US009096703B2

(12) United States Patent (10) Patent No.: US 9,096,703 B2 Li et al. (45) Date of Patent: Aug. 4, 2015

(54) NON-FOULING, ANTI-MICROBIAL, (56) References Cited ANT-THROMBOGENC GRAFT FROM COMPOSITIONS U.S. PATENT DOCUMENTS 4,098,728 A 7, 1978 Rosenblatt (75) Inventors: Jun Li, Cambridge, MA (US); Zheng 4,211,227 A 7, 1980 Anderson et al. 4,636,208 A 1, 1987 Rath Zhang, Cambridge, MA (US); Chad C. 4,877,864 A 10/1989 Wang et al. Huval, Cambridge, MA (US); Michael 5,013,649 A 5/1991 Wang et al. A. Bouchard, Cambridge, MA (US); 5, 180,375 A 1/1993 Feibus Arthur J. Coury, Cambridge, MA (US); 5,453,467 A * 9/1995 Bamford et al...... 525/287 Christopher R. Loose, Cambridge, MA 5,661,007 A 8/1997 Wozney et al. 5,688,678 A 11/1997 Hewicket al. (US) 5,739,236 A 4/1998 Bowers et al. 5,844,016 A 12/1998 Sawhney et al. 5,866,113 A 2f1999 Hendriks et al. (73) Assignee: SEMPRUS BIOSCIENCES 5.997,895 A 12/1999 Narotam et al. CORPORATION, Cambridge, MA 6,054,504 A 4/2000 Dalla Riva Toma (US) 6,150,459 A 1 1/2000 Mayes et al. 6,177,406 B1 1/2001 Wang et al. 6,251,964 B1 6/2001 Porssa et al. (*) Notice: Subject to any disclaimer, the term of this 6,358,557 B1 3/2002 Wang et al. patent is extended or adjusted under 35 6,395,800 B1 5, 2002 Jones et al. 6.432,919 B1 8/2002 Wang et al. U.S.C. 154(b) by 309 days. 6,489,382 B1 12/2002 Giesecke et al. 6,534,268 B1 3/2003 Kawai et al. 6,558,734 B2 5/2003 Koulik et al. (21) Appl. No.: 13/156,677 6,559,242 B1 5/2003 Ball et al. 6,589,665 B2 7/2003 Chabrecek et al. 6,711,879 B2 3/2004 Korteweg et al. (22) Filed: Jun. 9, 2011 7,087,658 B2 8, 2006 Swan et al. 7,220,491 B2 5/2007 Rounset al. (65) Prior Publication Data 7,238.364 B2 7/2007 Sawhney et al. 7,238,426 B2 7/2007 Jiang et al. US 2011/0305872A1 Dec. 15, 2011 7.276.286 B2 10/2007 Chapman et al. 7,306.625 B1 12/2007 Stratford et al. (Continued) Related U.S. Application Data FOREIGN PATENT DOCUMENTS (60) Provisional application No. 61/353,059, filed on Jun. 9, 2010. WO O3,OOO433 A1 1, 2003 WO 2007/OO2493 A2 1, 2007 (Continued) (51) Int. Cl. C08F 255/02 (2006.01) OTHER PUBLICATIONS A6IL 29/06 (2006.01) A6IL 29/08 (2006.01) Odian, George, Principles of Polymerization, 1991, John Wiley & A6IL 33/00 (2006.01) Sons Inc., 3rd Edition, pp. 6-7.* CSF 29/00 (2006.01) (Continued) C08. 7/16 (2006.01) C08. 7/8 (2006.01) C08G 18/83 (2006.01) Primary Examiner — Aaron Austin C08G 64/42 (2006.01) Assistant Examiner — Jasper Saberi C08G 77/38 (2006.01) (74) Attorney, Agent, or Firm — Bryan Cave LLP (52) U.S. Cl. CPC ...... C08F 255/02 (2013.01); A61L 29/06 (57) ABSTRACT (2013.01); A61L 29/085 (2013.01); A61L 33/0088 (2013.01); C08F 291/00 (2013.01); A method for preparing and resulting articles of manufacture C08G 18/836 (2013.01); C08G 64/42 comprising a Substrate having a Surface, a bulk beneath the (2013.01); C08G 77/38 (2013.01); C08J 7/16 Surface, and a grafted polymer layer on the Substrate surface, (2013.01); C08J 7/18 (2013.01); C08.J 2323/12 the Substrate surface and the grafted polymer layer, in com (2013.01); C08J 2369/00 (2013.01); C08.J. bination, constituting a modified Surface having a fibrinogen 2383/04 (2013.01); C08J 2433/14 (2013.01); adsorption of less than about 125 ng/cm in a fibrinogen Y10T 428/24355 (2015.01); Y10T 428/265 binding assay in which the modified surface is incubated for (2015.01) 60 minutes at 37° C. in 70 g/mL fibrinogen derived from human plasma containing 1.4 ug/mL I-125 radiolabeled (58) Field of Classification Search fibrinogen. CPC. A61L 33/00; A61L 33/0088; Y1OT 428/265; CO8F 255/02 See application file for complete search history. 25 Claims, No Drawings US 9,096,703 B2 Page 2

(56) References Cited Their Ability to Resist Protein Adsorption. The Journal of Physical Chemistry B 1998, 102 (2), 426-436. U.S. PATENT DOCUMENTS Haynie, S. L.; Crum, G. A.; Doele, B. A., Antimicrobial activities of amphiphilic peptides covalently bonded to a water-insoluble resin. 7,431,888 B2 10/2008 Frechet et al. 2001/0050749 A1 12/2001 Watanabe Antimicrobial Agents and Chemotherapy 1995, 39 (2), 301-307. 2003/0021823 A1 1/2003 Landers et al. Ignatova et al., Combination of electrografting and atom-transfer 2003.0143335 A1 7/2003 Qiu et al. radical polymerization for making the stainless steel Surface antibac 2004/0253383 A1 12/2004 Belik et al. terial and protein antiadhesive, Langmuir, 2005, 22(1), 255-262. 2004/O256232 A1 12/2004 Jiang et al. Ishihara, K.; Iwasaki, Y.; Ebihara, S.; Shindo, Y.; Nakabayashi, N., 2006/0057180 A1* 3/2006 Chilkoti et al...... 424/422 Photoinduced graft polymerization of 2-methacryloyloxyethyl 2006/0217285 A1 9, 2006 DeStarac phosphorylcholine on polyethylene membrane Surface for obtaining 2007/004.8249 A1 3/2007 Youngblood et al. 2007,025.4006 A1 11/2007 Loose et al. blood cell adhesion resistance. Colloids Surf B Biointerfaces 2000, 2008/0181861 A1* 7/2008 Jiang et al...... 424/78.09 18 (3-4), 325-335. 2009. O155335 A1 6/2009 O'Shaughnessey et al. Yuan, J.; Zhang, J.; Zang, X. Shen, J.; Lin, S., Improvement of blood 2009/0162662 A1* 6/2009 Chang et al...... 428/421 compatibility on cellulose membrane Surface by grafting betaines. 2009/O197791 A1 8, 2009 BalaStreet al. Colloids and Surfaces B: Biointerfaces 30, 2003. 2010, 0072642 A1 3/2010 Broad et al. Jiang, Y.; Rongbing, B., Ling, T., Jian, S.; Sicong, L., Blood compat 2010.009916O A1* 4/2010 Jiang et al...... 435/180 ibility of polyurethane Surface grafted copolymerization with Sulfobetaine monomer. Colloids Surf B Biointerfaces 2004, 36 (1), FOREIGN PATENT DOCUMENTS 27-33. WO 2007/024393 A2 3, 2007 Jin, Z.; Feng, W.; Beisser, K. Zhu, S.; Sheardown, H.; Brash, J. L., WO 2007/095393 A2 8, 2007 Protein-resistant polyurethane prepared by Surface-initiated atom WO 2008/006911 A1 1, 2008 transfer radical graft polymerization (ATRgP) of water-soluble poly WO 2008/O19381 A2 2, 2008 mers: effects of main chain and side chain lengths of grafts. Colloids WO 2008.083390 A2 T 2008 Surf B Biointerfaces 2009, 70 (1), 53-9. WO 2009/085096 A2 T 2009 Jin, Z.; Feng, W.; Zhu, S.; Sheardown, H.; Brash, J. L., Protein resistant polyurethane via Surface-initiated atom transfer radical OTHER PUBLICATIONS polymerization of oligo(ethylene glycol) methacrylate. J Biomed Mater Res A 2009, 91 (4), 1189-201. Salim, M. et al., Studies of electroosmotic flow and the effects of Zhang, J.; Yuan, J.; Yuan, Y.; Shen, J., Lin, S., Chemical modification protein adsorption in plasma-polymerized microchannel Surfaces, of cellulose membranes with Sulfo ammonium Zwitterionic vinyl Electrophoresis 2009, 30, 1877-1887. monomer to improve hemocompatibility. Colloids and Surfaces B: Patent Cooperation Treaty, International Search Report for PCT/ Biointerfaces 30. US 11/39790, mailed Mar. 12, 2012, 4 pages. Jun, Z. Youling, Y.; Kehua, W.; Jian, S.; Sicong, L., Surface modifi Patent Cooperation Treaty, International Search Report for PCT/ cation of segmented poly(ether urethane) by grafting Sulfo ammo US 11/39792, mailed Mar. 12, 2012, 6 pages. nium Zwitterionic monomer to improve hemocompatibilities. Col Cheng, N. et al. Thickness-Dependent Properties of PolyZwit loids and Surfaces B: Biointerfaces 2003, 28 (1), 1-9. terionic Brushes, Macromolecules, 2008, 41(17), 6317-6321. Kang, E.T.: Tan, K. L.; Liaw, D. J.; Chiang, H. H., Surface modifi Weinstock, B.A., et al., Rapid, Nondestructive Estimation of Surface cation and functionalization of electroactive polymer films via graft Polymer Layer Thickness. Using Attenuated Total Reflection Fourier ing of polyelectrolyte, polyampholyte and polymeric acids. Journal Transform Infrared (ATR FT-IR) Spectroscopy and Synthetic Spec to Materials Science 1996, 31, 1295-1301. tra Derived from Optical Principles, Applied Spectroscopy, 2012, Keiji Fujimoto. Y.T., Hiroyuki Inoue, Yoshito Ikada, Ozone-induced 66(11) 1311-1319. graft polymerization onto polymer surface. J Polym Sci A Polym Odian, G., Polymerization Mechanism. Types of Polymers and Poly Chem 1993, 31, 1035-1043. merizations, p. 6-7. Kildal, K.; Olafsen, K.; Stori, A., Peroxide-initiated grafting of Bell, C.; Peppas, N., Biomedical membranes from hydrogels and acrylamide on to polyethylene Surfaces. Journal of Applied Polymer interpolymer complexes. Biopolymers II 1995, 122, 125-175. Science 1992, 44 (11), 1893-1898. Chapman, R. G.; Ostuni, E.; Liang, M. N.; Meluleni, G.; Kim, E.; Wang, L.; Su, Y-I.; Zheng, L.; Chen, W.; Jiang, Z. Highly efficient Yan. L.; Pier, G.; Warren, H. S.; Whitesides, G. M., Polymeric Thin antifouling ultrafiltration membranes incorporating Zwitterionic Films That Resist the Adsorption of Proteins and the Adhesion of poly(3-(methacryloylamino)propyl-dimethyl(3-sulfopropyl) Bacteria. Langmuir 2001, 17 (4), 1225-1233. ammonium hydroxide). Journal of Membrane Science 340, 2009. Cheng, G.; Li, G.; Xue, H.; Chen, S.; Bryers, J. D.; Jiang, S. Zwit Liu, P-S.; Chen, Q.: Liu, X.;Yuan, B.; Wu, S.-S.; Shen, J.; Lin, S.-C., terionic carboxybetaine polymer Surfaces and their resistance to Grafting of Zwitterion from Cellulose Membranes via ATRP for long-term biofilm formation. Biomaterials 2009, 30 (28), 5234-40. Improving Blood Compatibility. Biomacromolecules 2009, 10 (10), Cheng. G.; Zhang, Z.; Chen, S.; Bryers, J. D., Jiang, S., Inhibition of 2809-2816. bacterial adhesion and biofilm formation on Zwitterionic Surfaces. Massia, S. P.; Stark, J., Immobilized RGD peptides on surface Biomaterials 2007, 28 (29), 4192-9. grafted dextran promote biospecific cell attachment. J Biomed Mater Du, H.; Chandaroy, P.; Hui, S.W. Grafted poly-(ethylene glycol) on Res 2001, 56 (3), 390-9. lipid surfaces inhibits protein adsorption and cell adhesion. Michel, R.; Pasche, S.; Textor, M. Castner, D. G., Influence of PEG Biochimica et Biophysica Acta (BBA)—Biomembranes 1997, 1326 Architecture on Protein Adsorption and Conformation. Langmuir (2), 236-248. 2005, 21 (26), 12327-12332. Feng, W.; Brash, J.; Zhu, S., Atom-Transfer Radical Grafting Poly Sakharov AM. M. L., Skibida IP. Catalytic oxidative deformylation merization of 2-Methacryloyloxyethyl Phosphorycholine from Sili of polyethylene glycols with the participation of molecular oxygen. con Wafer Surfaces. Journal of Polymer Science Part A: Polymer Kinet Catal 2001, 42, 662-668. Chemistry 2004, 42, 2931-2942. Villa-Diaz, L. G.; Nandivada, H.; Ding, J.; Nogueira-de-Souza, N. Goda, T., Konno, T.: Takai, M.; Moro, T.: Ishihara, K., Biomimetic C.; Krebsbach, P. H.; O'Shea, K. S.; Lahann, J.; Smith, G. D., Syn phosphorylcholine polymer grafting from polydimethylsiloxane Sur thetic polymer coatings for long-term growth of human embryonic face using photo-induced polymerization. Biomaterials 2006, 27 stem cells. Nat Biotechnol 2010, 28 (6), 581-3. (30), 5151-60. West, S. L.; Salvage, J. P. Lobb, E.J.; Armes, S. P.; Billingham, N.C.; Harder, P.; Grunze, M.; Dahint, R.; Whitesides, G. M.; Laibinis, P. E., Lewis, A. L.; Hanlon, G. W.; Lloyd, A. W., The biocompatibility of Molecular Conformation in Oligo(ethylene glycol)-Terminated Self crosslinkable copolymer coatings containing Sulfobetaines and Assembled Monolayers on Gold and Silver Surfaces Determines phosphobetaines. Biomaterials 2004, 25 (7-8), 1195-1204. US 9,096,703 B2 Page 3

(56) References Cited to improve hemocompatibility. Colloids and Surfaces B: Biointerfaces 2003, 29, 247-256. OTHER PUBLICATIONS Zhang, V., Cheng, Yang, Xue, Jiang. Nonfouling Behavior of Polycarboxybetaine-Grafted Surfaces: Structural and Environmental Wozney, J. M.; Rosen, V.: Celeste, A.J.; Mitsock, L. M.; Whitters, M. Effects—Biomacromolecules (ACS Publications). J.; Kriz, R. W.; Hewick, R. M.; Wang, E. A., Novel regulators of bone Biomacromolecules (Web): Sep. 12, 2008, 10, 2686-92. formation: molecular clones and activities. Science 1988, 242 Zhang, Z.; Chen, S.; Chang.Y.; Jiang, S., Surface grafted Sulfobetaine (4885), 1528-1534. Yuan, Y.; Ai, F. Zang, X. Zhuang, W.; Shena, J., Lin, S., polymers via atom transfer radical polymerization as Superlow foul Polyurethane vascular catheter surface grafted with Zwitterionic ing coatings. J Phys Chem B 2006, 110 (22), 10799-804. Sulfobetaine monomer activated by ozone. Colloids and Surfaces B: Zhang, Z., Zhang, M., Chen, S.; Horbett, T. A.; Ratner, B. D., Jiang, Biointerfaces 35, 2004. S., Blood compatibility of surfaces with Superlow protein adsorption. Yuan, J.; Chen, L., Jiang, X. Shen, J.; Lin, S., Chemical graft poly Biomaterials 2008, 29 (32), 4285-91. merization of Sulfobetaine monomer on polyurethane Surface for Zhang, Z.; Chao, T., Chen, S.; Jiang, S., Superlow Fouling reduction in platelet adhesion. Colloids Surf B Biointerfaces 2004, Sulfobetaine and Carboxybetaine Polymers on Glass Slides. 39(1-2), 87-94. Langmuir 2006, 22 (24), 10072-10077. Yuan, J.; Zhang, J.; Zhou, J.;Yuan, Y. L.; Shen, J.; Lin, S.C., Platelet Jiang, Zwitterionic Separation Materials for Liquid Chromatography adhesion onto segmented polyurethane Surfaces modified by and Capillary Electrophoresis Synthesis, Characterization and Appli carboxybetaine. J Biomater Sci Polym Ed 2003, 14 (12), 1339-49. cation for Inorganic Ion and Biomolecule Separations, PhD Disser Yuan, Y.; Ai. F.; Zhang, X.; Shen, J.; Lin, S. Grafting Sulfobetaine tation, Umeå University, Umeå, Sweden, 63 pages, 2003. monomer onto the segmented poly(ether-urethane) Surface to International Search Report issued in PCT/US09/67013 on Jun. 14. improve hemocompatibility. J Biomaterial Sci Polym Ed 2002, 13, 2010, 4 pages. 1081-92. Yuan.Y.; Zhang, J.; Ai, F.Yuan, J.; Zhou, J.; Shen, J.; Lin, S., Surface modification of SPEC films by ozone induced graft copolymerization * cited by examiner US 9,096,703 B2 1. 2 NON-FOULING, ANTI-MICROBIAL, geously, the polymeric material may possess a range of poly ANT-THROMBOGENC GRAFT FROM meric backbones and substituents while providing the articles COMPOSITIONS with a highly efficient, biocompatible, and non-fouling Sur face. In another embodiment, bioactive compositions are CROSS-REFERENCE TO RELATED attached to the modified surface. APPLICATIONS One aspect of the present invention is the provision of non-fouling polymeric materials for various Substrates, such This application claims priority to U.S. Patent Application as polymers and metal oxides, which retain their activity in Ser. No. 61/353,059, filed Jun. 9, 2010, the contents of which the presence of blood proteins and/or in vivo due to improved are incorporated herein by reference in its entirety. 10 molecular structures. In one embodiment, bioactive compo sitions are attached to the non-fouling material. FIELD OF THE INVENTION Another aspect of the present invention is the provision of non-fouling compositions containing a high density of non The present invention generally relates to articles of manu 15 fouling polymeric material and/or wherein the inter-polymer facture, Such as medical devices, having a non-fouling Sur chain distance of the non-fouling polymeric materials face comprising a polymeric material that is grafted from the decreases the penetration of fouling molecules into the non article. The surface resists the adhesion of biological material. fouling polymer layer. BACKGROUND OF THE INVENTION A further aspect of the present invention is the provision of grafted polymer layers for medical devices or other articles Many different materials have been investigated to resist that are hydrophilic, but possess somewhat limited Swelling non-specific protein adsorption. Chemistries utilized for this capacity in water. purpose include, but are not limited to: polyethers (e.g., poly A further aspect of the present invention is the provision of ethylene glycol), polysaccharides Such as dextran, hydro graft-from methods for modifying a Surface of an article of philic polymers such as polyvinylpyrrolidone or hydroxy 25 manufacture wherein the grafting is initiated from the article ethyl-methacrylate, heparin, intramolecular Zwitterions or itself to provide the article with a polymeric grafted polymer mixed charge materials, and hydrogen bond accepting groups layer that is relatively thick and relatively uniformly distrib such as those described in U.S. Pat. No. 7,276,286. The ability uted on the Surface of the article. In general, the resulting of these materials in preventing protein adsorption varies polymeric grafted polymer layers are generally thicker than greatly between the chemistries. Of these materials, only a 30 self-assembled monolayer-initiated coatings and thus more few resist fouling to the degree required for short-term in vivo fully cover the defects and irregularities in commercial bio application. However, the few materials appropriate for short materials, including polymers and metals, so that non-fouling term application, when used for longer periods of time in grafted polymer layers are effective in complex media and/or complex media or in vivo, exhibit significant fouling or other in vivo. degradation, making them unsuitable for long-term applica 35 Briefly, therefore, one aspect of the present invention is an tions. Furthermore, surfaces coated with materials that resist article of manufacture comprising a polymeric Substrate hav in Vivo degradation are often susceptible to a noticeable ing a surface and a polymer layer on the Substrate Surface. The decrease in fouling resistance over time. Substrate surface and the polymer layer, in combination, con WO 2007/02493 describes grafting sulfobetaine and car stitute a modified Surface having a fibrinogen adsorption of boxybetaine from self-assembled monolayers on gold Sub 40 less than about 125 ng/cm in a fibrinogen binding assay in strates or from silyl groups on glass Substrates using atom which the modified surface is incubated for 60 minutes at 37° transfer radical polymerization (ATRP). Gold and glass are C. in a solution containing 70 ug/mL fibrinogen derived from not appropriate substrates for many medical devices used in human plasma and 1.4 ug/mL I-125 radiolabeled fibrinogen. vivo. Self-assembled monolayers, such as thiol-based mono In one embodiment, the Substrate surface and the polymer layers, may be unstable since the thiol group is not stably 45 layer, in combination, constitute a modified Surface having a bound to the substrate. fibrinogen adsorption of less than about 90 ng/cm in a U.S. Pat. No. 6,358,557 to Wang et al. describes the graft fibrinogen binding assay in which the modified Surface is polymerization of Substrate surfaces, but not with a high incubated for 60 minutes at 37°C. in a solution containing 70 density of a highly non-fouling polymeric material. Athermal ug/mL fibrinogen derived from human plasma and 1.4 ug/mL initiator is used to initiate polymerization, typically at tem 50 I-125 radiolabeled fibrinogen. In another embodiment, the peratures greater than 85°C. Such temperatures are generally Substrate surface and the polymer layer, in combination, con not suitable for many medical devices. Such as thin-walled stitute a modified Surface having a fibrinogen adsorption of polyurethane catheters. Further, the “salt out' method less than about 90 ng/cm in a fibrinogen binding assay in described is generally not suitable for grafting polymers such which the modified surface is incubated for 60 minutes at 37° as Zwitterionic polymers. 55 C. in a solution containing 70 ug/mL fibrinogen derived from Jian et al., Colloids and Surfaces B: Biointerfaces 28, 1-9 human plasma and 1.4 ug/mL I-125 radiolabeled fibrinogen. (2003) describes the surface modification of segmented poly In another embodiment, the substrate surface and the polymer (ether urethane) by grafting Sulfobetaine Zwitterionic mono layer, in combination, constitute a modified Surface having a mer, but not with a high density of non-fouling material. The fibrinogen adsorption of less than about 50 ng/cm in a resulting materials are not sufficiently non-fouling to be use 60 fibrinogen binding assay in which the modified Surface is ful in medical device applications. incubated for 60 minutes at 37°C. in a solution containing 70 ug/mL fibrinogen derived from human plasma and 1.4 ug/mL SUMMARY OF THE INVENTION I-125 radiolabeled fibrinogen. The present invention is further directed to an article of Among the various aspects of the present invention is the 65 manufacture comprising a polymeric Substrate having a Sur provision of medical devices and other articles having a non face and a layer of a graft-from polymer on the Substrate fouling polymeric material grafted therefrom. Advanta Surface. The Substrate Surface and the graft-from polymer, in US 9,096,703 B2 3 4 combination, constitute a modified surface having a static C. in a solution containing 70 ug/mL fibrinogen derived from contact angle of less than 25 degrees. human plasma and 1.4 ug/mL I-125 radiolabeled fibrinogen. The present invention is further directed to an article of The present invention is further directed to an article of manufacture comprising a substrate having a Surface and a manufacture comprising a substrate having a surface and a polymer layer on the substrate surface. The substrate surface polymer layer on the substrate surface. The substrate surface and the polymer layer, in combination, constitute a modified and the polymer layer, in combination, constitute a modified Surface having a fibrinogen adsorption of less than about 125 Surface having a fibrinogen adsorption of less than about 125 ng/cm in a fibrinogen binding assay in which the modified ng/cm in a fibrinogen binding assay in which the modified surface is incubated for 60 minutes at 37° C. in a solution surface is incubated for 60 minutes at 37° C. in a solution 10 containing 70 ug/mL fibrinogen derived from human plasma containing 70 ug/mL fibrinogen derived from human plasma and 1.4 ug/mL I-125 radiolabeled fibrinogen. The polymer and 1.4 g/mL I-125 radiolabeled fibrinogen. The polymer layer has a global average dry thickness that is at least equal layer has a global average dry thickness of at least about 50 to the global average R. Surface roughness of the Substrate nm and the magnitude of the difference between the global Surface. In one such embodiment, the Substrate surface and average dry thickness of the polymer layer as determined by 15 the polymer layer, in combination, constitute a modified Sur scanning electron microscopy (SEM) and the global average face having a fibrinogen adsorption of less than about 90 humidified thickness of the polymer layer as determined by ng/cm in a fibrinogen binding assay in which the modified environmental scanning electron microscopy (ESEM) is less surface is incubated for 60 minutes at 37° C. in a solution than 200% of the global average dry thickness. In one such containing 70 ug/mL fibrinogen derived from human plasma embodiment, the Substrate surface and the polymer layer, in and 1.4 ug/mL I-125 radiolabeled fibrinogen. In another such combination, constitute a modified Surface having a fibrino embodiment, the Substrate Surface and the polymer layer, in gen adsorption of less than about 90 ng/cm in a fibrinogen combination, constitute a modified Surface having a fibrino binding assay in which the modified surface is incubated for gen adsorption of less than about 70 ng/cm in a fibrinogen 60 minutes at 37° C. in a solution containing 70 ug/mL binding assay in which the modified surface is incubated for fibrinogen derived from human plasma and 1.4 g/mL I-125 25 60 minutes at 37° C. in a solution containing 70 ug/mL radiolabeled fibrinogen. In another such embodiment, the fibrinogen derived from human plasma and 1.4 ug/mL I-125 Substrate surface and the polymer layer, in combination, con radiolabeled fibrinogen. In another such embodiment, the stitute a modified Surface having a fibrinogen adsorption of Substrate surface and the polymer layer, in combination, con less than about 70 ng/cm in a fibrinogen binding assay in stitute a modified Surface having a fibrinogen adsorption of which the modified surface is incubated for 60 minutes at 37° 30 less than about 50 ng/cm in a fibrinogen binding assay in C. in a solution containing 70Lug/mL fibrinogen derived from which the modified surface is incubated for 60 minutes at 37° human plasma and 1.4 ug/mL I-125 radiolabeled fibrinogen. C. in a solution containing 70 ug/mL fibrinogen derived from In another such embodiment, the substrate surface and the human plasma and 1.4 ug/mL I-125 radiolabeled fibrinogen. polymer layer, in combination, constitute a modified Surface The present invention is further directed to an article of having a fibrinogen adsorption of less than about 50ng/cm in 35 manufacture comprising a substrate having a surface and a a fibrinogen binding assay in which the modified Surface is polymer layer on the substrate surface. The substrate surface incubated for 60 minutes at 37°C. in a solution containing 70 and the polymer layer, in combination, constitute a modified ug/mL fibrinogen derived from human plasma and 1.4 ug/mL Surface having a fibrinogen adsorption of less than about 125 I-125 radiolabeled fibrinogen ng/cm in a fibrinogen binding assay in which the modified The present invention is further directed to an article of 40 surface is incubated for 60 minutes at 37° C. in a solution manufacture comprising a substrate having a Surface and a containing 70 ug/mL fibrinogen derived from human plasma polymer layer on the substrate surface. The substrate surface and 1.4 ug/mL I-125 radiolabeled fibrinogen. In one embodi and the polymer layer, in combination, constitute a modified ment, the polymer layer has a global average R. Surface Surface having a fibrinogen adsorption of less than about 125 roughness that is less than 300% of the global average R. ng/cm in a fibrinogen binding assay in which the modified 45 Surface roughness of the Substrate Surface. In one embodi surface is incubated for 60 minutes at 37° C. in a solution ment, the polymer layer has a global average R. Surface containing 70 ug/mL fibrinogen derived from human plasma roughness that is less than the 200% of the global average and 1.4 ug/mL I-125 radiolabeled fibrinogen. The article R. Surface roughness of the Substrate Surface. In another further comprises a solvent extractable polymerization initia embodiment, the polymer layer has a global average R. tor or degradation product thereof. In one Such embodiment, 50 surface roughness that is less than 150% of the global average the Substrate Surface and the polymer layer, in combination, R. Surface roughness of the Substrate Surface. In another constitute a modified surface having a fibrinogen adsorption embodiment, the polymer layer has a global average R. of less than about 90 ng/cm in a fibrinogen binding assay in Surface roughness that is less than the global average R. which the modified surface is incubated for 60 minutes at 37° Surface roughness of the Substrate Surface. In each of the C. in a solution containing 70Lug/mL fibrinogen derived from 55 forgoing embodiments, the Substrate Surface and the polymer human plasma and 1.4 ug/mL I-125 radiolabeled fibrinogen. layer, in combination, may constitute a modified Surface hav In another such embodiment, the substrate surface and the ing a fibrinogen adsorption of less than about 90 ng/cm in a polymer layer, in combination, constitute a modified Surface fibrinogen binding assay in which the modified Surface is having a fibrinogen adsorption of less than about 70ng/cm in incubated for 60 minutes at 37°C. in a solution containing 70 a fibrinogen binding assay in which the modified Surface is 60 ug/mL fibrinogen derived from human plasma and 1.4 ug/mL incubated for 60 minutes at 37°C. in a solution containing 70 I-125 radiolabeled fibrinogen. In each of the forgoing ug/mL fibrinogen derived from human plasma and 1.4 ug/mL embodiments, the Substrate Surface and the polymer layer, in I-125 radiolabeled fibrinogen. In another such embodiment, combination, constitute a modified Surface having a fibrino the Substrate Surface and the polymer layer, in combination, gen adsorption of less than about 70 ng/cm in a fibrinogen constitute a modified surface having a fibrinogen adsorption 65 binding assay in which the modified surface is incubated for of less than about 50 ng/cm in a fibrinogen binding assay in 60 minutes at 37° C. in a solution containing 70 ug/mL which the modified surface is incubated for 60 minutes at 37° fibrinogen derived from human plasma and 1.4 ug/mL I-125 US 9,096,703 B2 5 6 radiolabeled fibrinogen. In each of the forgoing embodi embodiment, the Substrate surface has a global average R. ments, the Substrate Surface and the polymer layer, in combi Surface roughness of at least 150 nm and graft polymerization nation, may constitute a modified Surface having a fibrinogen is continued until the polymer has a global average dry thick adsorption of less than about 50 ng/cm in a fibrinogen bind ness that exceeds the global average R. Surface roughness ing assay in which the modified surface is incubated for 60 of the substrate surface. In another such embodiment, the minutes at 37°C. in a solution containing 70 ug/mL fibrino Substrate Surface has a global average R. Surface roughness gen derived from human plasma and 1.4 ug/mL I-125 radio of at least 100 nm and graft polymerization is continued until labeled fibrinogen. For example, in one embodiment, the the polymer has a global average dry thickness that exceeds Substrate surface and the polymer layer, in combination, con the global average R. Surface roughness of the substrate stitute a modified Surface having a fibrinogen adsorption of 10 Surface. In another such embodiment, the Substrate Surface less than about 50 ng/cm in a fibrinogen binding assay in has a global average R. Surface roughness of at least 50 nm which the modified surface is incubated for 60 minutes at 37° and graft polymerization is continued until the polymer has a C. in a solution containing 70Lug/mL fibrinogen derived from global average dry thickness that exceeds the global average human plasma and 1.4 g/mL I-125 radiolabeled fibrinogen R, surface roughness of the substrate surface. and the polymer layer has a global average R. Surface 15 For articles such as microfluidic devices and wovenmeshes roughness that is less than the global average R. Surface which are designed to have channels or pores having a size roughness of the Substrate surface. (e.g., diameter or width) in the range of 100 nm to 1 millime The present invention is further directed to an article of ter, it may be desired to have a global average dry thickness of manufacture comprising a substrate having a Surface and a the graft polymer layer that is less than 10% of the channel or polymer layer on the substrate surface. The substrate surface pore size of the device. Without being bound by any theory, and the polymer layer, in combination, constitute a modified having a global average dry thickness Substantially smaller Surface having a fibrinogen adsorption of less than about 125 than the channel or pore size may reduce impact on the ng/cm in a fibrinogen binding assay in which the modified function of the device. In certain embodiments, it is desired to surface is incubated for 60 minutes at 37° C. in a solution have a global average dry thickness of the graft polymer layer containing 70 ug/mL fibrinogen derived from human plasma 25 that is less than 5% of the channel or pore size of the device. and 1.4 g/mL I-125 radiolabeled fibrinogen. The polymer In certain embodiments, it is desired to have a global average layer has a global average dry thickness wherein the standard dry thickness of the graft polymer layer that is less than 3% of deviation of the global average dry thickness of the polymer the channel or pore size of the device. In certain embodi layer does not exceed 100% of the global average dry thick ments, it is desired to have a global average dry thickness of ness of the polymer layer. In one such embodiment, the Sub 30 the graft polymer layer that is less than 1% of the channel or strate Surface and the polymer layer, in combination, consti pore size of the device. In certain embodiments, it is desired tute a modified surface having a fibrinogen adsorption of less to have a global average dry thickness of the graft polymer than about 90 ng/cm in a fibrinogen binding assay in which layer that is less than 0.1% of the channel or pore size of the the modified surface is incubated for 60 minutes at 37°C. in device. a solution containing 70 ug/mL fibrinogen derived from 35 The present invention is further directed to a process for human plasma and 1.4 ug/mL I-125 radiolabeled fibrinogen. grafting a polymer from an article comprising a substrate In another such embodiment, the substrate surface and the having a Surface, a bulk beneath the Surface, and a near polymer layer, in combination, constitute a modified Surface surface Zone lying between the surface and the bulk. The having a fibrinogen adsorption of less than about 70 ng/cm in process comprises incorporating a polymerization initiator a fibrinogen binding assay in which the modified Surface is 40 into the near-Surface Zone and graft polymerizing a polymer incubated for 60 minutes at 37°C. in a solution containing 70 from the Substrate surface to form a polymer layer comprising ug/mL fibrinogen derived from human plasma and 1.4 ug/mL the grafted polymer, the polymer layer having a global aver I-125 radiolabeled fibrinogen. In another such embodiment, age dry thickness of at least about 200 nm. The magnitude of the Substrate Surface and the polymer layer, in combination, the difference between the global average dry thickness of the constitute a modified surface having a fibrinogen adsorption 45 polymer layer as determined by scanning electron micros of less than about 50 ng/cm in a fibrinogen binding assay in copy (SEM) and the global average humidified thickness of which the modified surface is incubated for 60 minutes at 37° the polymer layer as determined by environmental scanning C. in a solution containing 70Lug/mL fibrinogen derived from electron microscopy (ESEM) is less than 200% of the global human plasma and 1.4 ug/mL I-125 radiolabeled fibrinogen. average dry thickness. In one Such embodiment, the polymer The present invention is further directed to a process for 50 layer has a global average dry thickness of at least about 150 grafting a polymer from an article of manufacture comprising nm and the magnitude of the difference between the global a Substrate having a Surface, a bulk beneath the Surface, a average dry thickness of the polymer layer as determined by near-surface Zone lying between the surface and the bulk. The scanning electron microscopy (SEM) and the global average process comprises incorporating a polymerization initiator humidified thickness of the polymer layer as determined by into the near-Surface Zone and graft polymerizing a polymer 55 environmental scanning electron microscopy (ESEM) is less from the article. than 200% of the global average dry thickness. In another The present invention is further directed to a process for Such embodiment, the polymer layer has a global average dry grafting a polymer from an article of manufacture comprising thickness of at least about 100 nm and the magnitude of the a Substrate having a Surface, a bulk beneath the Surface, a difference between the global average dry thickness of the near-surface Zone lying between the surface and the bulk. The 60 polymer layer as determined by scanning electron micros process comprises incorporating a polymerization initiator copy (SEM) and the global average humidified thickness of into the near-Surface Zone and graft polymerizing a polymer the polymer layer as determined by environmental scanning from the article wherein the substrate surface has a global electron microscopy (ESEM) is less than 200% of the global average R. Surface roughness of at least 200 nm and graft average dry thickness. In another Such embodiment, the poly polymerization is continued until the polymer has a global 65 mer layer has a global average dry thickness of at least about average dry thickness that exceeds the global average R. 50 nm and the magnitude of the difference between the global Surface roughness of the Substrate surface. In one Such average dry thickness of the polymer layer as determined by US 9,096,703 B2 7 8 scanning electron microscopy (SEM) and the global average charged State or in a non-charged state, but in the non-charged humidified thickness of the polymer layer as determined by state is capable of becoming negatively charged, e.g., upon environmental scanning electron microscopy (ESEM) is less removal of an electrophile (e.g., a proton (H+), for example in than 200% of the global average dry thickness. a pH dependent manner) or a protecting group (e.g., a car The present invention is further directed to a process for 5 boxylic acid ester), or the addition of a nucleophile. In certain grafting a polymer from an article comprising a substrate instances, the group is Substantially negatively charged at an having a Surface, a bulk beneath the Surface, and a near approximately physiological pH but undergoes protonation surface Zone lying between the surface and the bulk. The and becomes substantially neutral at a weakly acidic pH. The process comprises incorporating a polymerization initiator non-limiting examples of Such groups include carboxyl and optionally other species such as ligands and/or catalysts 10 groups, barbituric acid and derivatives thereof, Xanthine and into the near-Surface Zone and graft polymerizing a polymer derivatives thereof, boronic acids, phosphinic acids, phos from the Substrate surface to form a polymer layer comprising phonic acids, sulfinic acids, Sulfonic acids, phosphates, and the grafted polymer, the polymer layer having a global aver Sulfonamides. age dry thickness that is at least equal to the global average Anionic species or Anionic moiety: unless otherwise indi R. Surface roughness of the Substrate surface. 15 cated, an “Anionic species' oran “Anionic moiety' is a group, Other objects and features will be in part apparent and in residue or molecule that is present in a negatively charged or part pointed out hereinafter. non-charged State, but in the non-charged state is capable of becoming negatively charged, e.g., upon removal of an elec ABBREVIATIONS AND DEFINITIONS trophile (e.g., a proton (H+), for example in a pH dependent manner) or other protecting group (e.g., a carboxylic acid The following definitions and methods are provided to ester), or the addition of a nucleophile. In certain instances, better define the present invention and to guide those of ordi the group, residue or molecule is substantially negatively nary skill in the art in the practice of the present invention. charged at an approximately physiological pH but undergoes Unless otherwise noted, terms are to be understood according protonation and becomes Substantially neutral at a weakly to conventional usage by those of ordinary skill in the relevant 25 acidic pH. art. Antibiofilm activity: unless otherwise indicated, “antibio When introducing elements of the present invention or the film activity” may be quantified, for example, using a stan preferred embodiment(s) thereof, the articles “a,” “an,” “the dard continuous flow assay. In one such assay, samples may and “said are intended to mean that there are one or more of be pre-incubated with 50% fetal bovine serum for 18-20 the elements. The terms “comprising.” “including and “hav 30 hours at 120 RPM at 37° C. Following preincubation, samples ing are intended to be inclusive and mean that there may be are then exposed to a subculture of bacteria via a modified additional elements other than the listed elements. CDC (mCDC) to make a bacterial suspension of 10°Cfu/mL Aliphatic: unless otherwise indicated, “aliphatic' or “ali in 1xRBS. The reactor is run in batch mode for 2 hours at 37° phatic group” means an optionally Substituted, non-aromatic C. with agitation. Thereafter, the samples are transferred to a hydrocarbon moiety. The moiety may be, for example, linear, 35 fresh reactor a suitable growth media for where flow of the branched, or cyclic (e.g., mono or polycyclic Such as fused, sterile media (8 mL/min) runs 20-23 hours with agitation. In bridging, or spiro-fused polycyclic), or a combination one preferred embodiment, the bacterial strain is Staphylo thereof. Unless otherwise specified, aliphatic groups contain coccus epidermidis (S. epidermidis, ATCC 35984), and the 1-20 carbon atoms. growth media used is 1:10 Tryptic soy broth (TSB)+0.25 wt Alkyl: unless otherwise indicated, the alkyl groups 40 % glucose. In an alternate preferred embodiment, the bacte described herein are preferably lower alkyl containing from rial strain is Escherichia coli (E. coli, ATCC 25922) and the one to eight carbon atoms in the principal chain and up to 20 growth media is M63 media supplemented with 1 mM carbon atoms. They may be linear, branched or cyclic and MgSO, 0.2% glucose, and 0.5% casamino acids. After incu include methyl, ethyl, propyl, butyl, hexyl and the like. bation, the samples are rinsed five times in 100 mL of 1xPBS Amino: unless otherwise indicated, the term "amino” as 45 to remove bacteria not tightly attached. Then, accumulated used herein alone or as part of another group denotes the bacteria on materials are macroscopically rated for biofilm moiety —NR'R' wherein R', and R are independently Surface coverage and are removed by Sonication in a new hydrogen, hydrocarbyl, substituted hydrocarbyl or heterocy solution of PBS and the total number of bacterial cells quan clo. tified through dilution plating. Preferably at least a 1, 2, 3 or Ammonium: unless otherwise indicated, the term "ammo 50 4 log reduction in bacterial count is found on the article with nium' as used herein alone or as part of another group denotes the non-fouling polymer layer relative to a reference Sub the moiety - N'R'R'R' wherein R, R and Rare indepen strate, that is, the same or an otherwise functionally equiva dently hydrogen, hydrocarbyl, substituted hydrocarbyl or lent Substrate lacking the non-fouling polymer layer. An heterocyclo. article that has a 1 log reduction in adhered bacteria relative to Amide or Amido: unless otherwise indicated, the "amide' 55 a reference substrate is said to have antibiofilm activity of 1 or "amido’ moieties represent a group of the formula log. An article that has a 2 log reduction in adhered bacteria —CONR'R' wherein R' and Rare as defined in connection relative to a reference substrate is said to have antibiofilm with the term “amino.” “Substituted amide.” for example, activity of 2 log, and so forth. refers to a group of the formula—CONR'R' wherein at least Antimicrobial: unless otherwise indicated, “antimicro one of R' and R are other than hydrogen. “Unsubstituted 60 bial refers to molecules and/or compositions that kill (i.e., amido.” for example, refers to a group of the formula microbicidal), inhibit the growth of (i.e., microbistatic), and/ —CONR'R'', wherein R' and Rare each hydrogen. or prevent fouling by, microorganisms including bacteria, Anionic Monomer, Anionic Monomeric Unit or Anionic yeast, fungi, mycoplasma, viruses or virus infected cells, Repeat Unit: unless otherwise indicated, an "anionic mono and/or protozoa. Antimicrobial activity with respect to bac mer.” "anionic monomeric unit' or “anionic repeat unit is a 65 teria may be quantified, for example, using a standard assay. monomer or monomeric unit bearing an anion or other In one such assay, samples may be pre-incubated with 50% anionic species, e.g., a group that is present in a negatively fetal bovine serum for 18-20 hours at 120 RPM at 37° C. US 9,096,703 B2 10 Following pre-incubation, samples are placed in Staphyllo nal amplifying molecule, including, but not limited to, a pro coccus aureus (S. aureus, ATCC 25923) which has been tein kinase, a cytokine, a chemokine, an interferon, tumor diluted from an overnight culture to a planktonic concentra necrosis factor, growth factor, growth factor inhibitor, hor tion of 1-3x10 CFU/mL in 1% tryptone soy broth (TSB) mone, enzyme, receptor-targeting ligand, gene silencing diluted in 1xRBS or other suitable media. Samples are incu agent, ambisense, antisense, an RNA, a living cell, cohesin, bated with bacteria for 24-26 hrs with agitation (120 rpm) at laminin, fibronectin, fibrinogen, osteocalcin, osteopontin, or 37°C. The concentration of TSB or other media can vary with osteoprotegerin. Bioactive agents can be aptamers, proteins, the organism being used. After incubation, the samples are glycoproteins, peptides, oligliopeptides, polypeptides, poly placed in 3 mL PBS for 5 min at 240 RPM at 37°C. to remove mers, inorganic compounds, organometallic compounds, bacteria not tightly attached to the material. Then, accumu 10 organic compounds or any synthetic or natural, chemical or lated bacteria on materials are removed by Sonication in a new biological compound. solution of PBS and the total number of bacterial cells are Biocompatibility: unless otherwise indicated, “biocompat quantified through dilution plating. Preferably at least a 1, 2, ibility” is the ability of a material to perform with an appro 3 or 4 log reduction in bacterial count occurs relative to priate host response in a specific situation. This can be evalu colonization on a reference Substrate, that is, the same or an 15 ated using International Standard ISO 10993. Biocompatible otherwise functionally equivalent Substrate lacking the non compositions described herein are preferably substantially fouling polymer layer. A surface that has a lower bacterial non-toxic. count on it than the reference substrate may be said to reduce Biological fluids: unless otherwise indicated, “biological microbial colonization. fluids' are fluids produced by organisms containing proteins Antimicrobial peptide (AmP): unless otherwise indicated, and/or cells, as well as fluids and excretions from microbes. “antimicrobial peptide' (or “AmP) refers to oligopeptides, This includes, but is not limited to, blood, saliva, urine, cere polypeptides, or peptidomimetics that kill (i.e., are microbi broSpinal fluid, tears, semen, lymph, ascites, sputum, bone cidal) or inhibit the growth of (i.e., are microbistatic) micro marrow, synovial fluid, aqueous humor, cerumen, organisms including bacteria, yeast, fungi, mycoplasma, broncheoalveolar lavage fluid, prostatic fluid, cowper's fluid viruses or virus infected cells, and/or protozoa. 25 or pre-ejaculatory fluid, Sweat, fecal matter, cyst fluid, pleural Anti-thrombogenic: unless otherwise indicated, “anti and peritoneal fluid, chyme, chyle, bile, intestinal fluid, pus, thrombogenic’ refers to the ability of a composition to resist sebum, Vomit, mucosal Secretion, stool water, pancreatic thrombus formation. Anti-thrombogenic activity can be juice, lavage fluids from sinus cavities, bronchopulmonary evaluated using an ex-Vivo flow loop model of thrombosis. aspirates, or any derivative thereof (e.g., serum, plasma). Briefly, up to 10 liters of fresh blood are collected from a 30 Block Copolymer: unless otherwise indicated, a “block single animal (bovine). This blood is heparinized to prevent copolymer comprises two or more homopolymer or copoly coagulation, filtered to remove particulates, and autologous mer subunits linked by covalent bonds. Block copolymers radio-labeled platelets are added. Within eight hours after with two or three distinct blocks are called diblock copoly blood harvesting, coated and uncoated articles are placed in a mers and triblock copolymers, respectively. A schematic gen flow loop circuit, which pumps blood from a bath over the 35 eralization of a diblock copolymer is represented by the for article and thenbackinto the bath. A second internal flow loop mula A,B,C...... -IX,Y,Z. . . . . wherein each letter circuit can be established for an article containing alumen by stands for a constitutional or monomeric unit, and wherein connecting the two ports of the article through a 2nd peristal each Subscript to a constitutional unit represents the mole tic pump. The size of tubing into which the article is placed fraction of that unit in the particular block, the three dots and speed of the blood flow may be adjusted based on the size 40 indicate that there may be more (there may also be fewer) of the article being tested. constitutional units in each block and m and n indicate the Aryl: unless otherwise indicated, the term “aryl' or “aryl molecular weight of each block in the diblock copolymer. As group' refers to optionally Substituted monocyclic, bicyclic, Suggested by the schematic, in some instances, the number and tricyclic ring systems having a total of five to fourteen and the nature of each constitutional unit is separately con ring members, wherein at least one ring in the system is 45 trolled for each block. The schematic is not meant and should aromatic and wherein each ring in the system contains three to not be construed to infer any relationship whatsoever between seven ring members. The terms “aryl or “ar as used herein the number of constitutional units or the number of different alone or as part of another group denote optionally Substituted types of constitutional units in each of the blocks. Nor is the homocyclic aromatic groups, preferably monocyclic orbicy schematic meant to describe any particular number or clic groups containing from 6 to 12 carbons in the ring por 50 arrangement of the constitutional units within a particular tion, such as phenyl, biphenyl, naphthyl, Substituted phenyl, block. In each block the constitutional units may be disposed substituted biphenyl or substituted naphthyl. Phenyl and sub in a purely random, an alternating random, a regular alternat stituted phenyl are the more preferred aryl. ing, a regular block or a random block configuration unless Attached: unless otherwise indicated, two moieties or expressly stated to be otherwise. A purely random configura compounds are “attached if they are held together by any 55 tion, for example, may have the non-limiting form: X-X- interaction including, by way of example, one or more cova Y—Z—X-Y Y-Z Y—Z—Z—Z. . . . A non-limiting, lent bonds, one or more non-covalent interactions (e.g., exemplary alternating random configuration may have the hydrogen bonds, ionic bonds, static forces, van der Waals non-limiting form: X Y X—Z Y X Y—Z Y interactions, combinations thereof, or the like), or a combi X-Z..., and an exemplary regular alternating configuration nation thereof. 60 may have the non-limiting form: X-Y-Z X-Y-Z- Bioactive Agent/Active Agent/Biomolecule: unless other X-Y-Z . . . . An exemplary regular block configuration wise indicated, “bioactive agent' or “active agent” or “bio may have the following non-limiting configuration: . . . molecule used herein synonymously, refers to any organic X X X Y Y Y-Z Z Z X—X X ..., while orinorganic therapeutic, prophylactic or diagnostic agent that an exemplary random block configuration may have the non actively or passively influences a biological system. For 65 limiting configuration: ... X-X-X-Z—Z X- X-Y- example, a bioactive agent can be an amino acid, antimicro Y Y Y-Z Z-Z X—X—Z—Z—Z— . . . . In a gra bial peptide, immunoglobulin, an activating, signaling or sig dient polymer, the content of one or more monomeric units US 9,096,703 B2 11 12 increases or decreases in a gradient manner from thea end of includes any increase in thickness to the Substrate or change the polymer to the () end. In none of the preceding generic in Surface chemical composition of the Substrate. examples is the particular juxtaposition of individual consti Complex Media: unless otherwise indicated, “complex tutional units or blocks or the number of constitutional units media' refers to biological fluids or Solutions containing pro in a block or the number of blocks meant nor should they be teins or digests of biological materials. Examples include, but construed as in any manner bearing on or limiting the actual are not limited to, cation-adjusted Mueller Hinton broth, tryp structure of block copolymers forming a micelle described tic soy broth, brain heart infusion, or any number of complex herein. As used herein, the brackets enclosing the constitu media, as well as any biological fluid. tional units are not meant and are not to be construed to mean Copolymer: unless otherwise indicated, “copolymer that the constitutional units themselves form blocks. That is, 10 refers to a polymer derived from two, three or more mono the constitutional units within the square brackets may com meric species and includes alternating copolymers, periodic bine in any manner with the other constitutional units within copolymers, random copolymers, statistical copolymers and the block, i.e., purely random, alternating random, regular block copolymers. alternating, regular block or random block configurations. Cysteine: unless otherwise indicated, "cysteine' refers to The block copolymers described hereinare, optionally, alter 15 the amino acid cysteine or a synthetic analogue thereof, nate, gradient or random block copolymers. In some embodi wherein the analogue contains a free Sulfhydryl group. ments, the block copolymers are dendrimer, star or graft Degradation Products: unless otherwise indicated, “degra copolymers. dation products are atoms, radicals, cations, anions, or mol Branched: unless otherwise indicated, “branched’ refers to ecules other than water formed as the result of hydrolytic, a polymer structure in which a polymer chain divides into two oxidative, enzymatic, or other chemical processes. or more polymer chains. Dry Thickness: unless otherwise indicated, “Dry Thick Brushes/Polymer Brushes: unless otherwise indicated, ness, as used herein in connection with a polymer layer, shall “brushes' or “polymer brushes are used herein synony mean the thickness of the polymer layer using a scanning mously and refer to polymerchains that are bound to a Surface electron microscope (SEM). To measure dry thickness, the generally through a single point of attachment using graft 25 sample is freeze fractured for imaging by being Submerged in from techniques. The polymers can be end-grafted (attached liquid nitrogen then cracked with an ultra microtome blade. via a terminal group) or attached via a side chain or a position For metal substrates, they may be scored with a notch before in the polymer chain other than a terminal position. The a primer or the non-fouling polymer is applied to make freeze polymers can be linear or branched. For example, the polymer fracturing easier. The freeze fracturing should break the chains described herein can contain a plurality of side chains 30 article at a plane approximately orthogonal to the polymer that contain Zwitterionic groups. The side chains can consist modified surface in order to measure the thickness of the of a single non-fouling moiety or monomer and/or a non polymer layer normal to the substrate. The samples are sput fouling oligomer (e.g., 2-10 monomeric residues) or polymer ter coated in gold for 90 seconds using a sputter coater and (e.g., >10 monomeric residues). then imaged underhigh vacuum at 5 kV using an SE2 detector Carboxyammonium: unless otherwise indicated, a "car 35 under a Field Emission Scanning Electron Microscope boxyammonium moiety is a Zwitterionic moiety comprising (SEM). Exemplary microtome blades include the Leica carboxylate and ammonium functionality and includes, for Ultracut UCT Ultramicrotome, exemplary sputter coaters example, carboxyammonium monomers, carboxyammo include the Cressington 208HR, exemplary SEMs include the nium oligomers, carboxyammonium polymers, carboxyam Supraš5VP FESEM, Zeiss. Dry thickness may be approxi monium repeat units, and other carboxyammonium-contain 40 mated by analyzing intensity of chemical signals in the ing materials. Carboxybetaine monomers, oligomers, grafted polymer, for instance, through the use of ATR-FTIR. polymers, repeat units and other carboxybetaine materials are Fibrinogen Adsorption Assay: unless otherwise indicated, exemplary carboxyammonium moieties. a “Fibrinogen Adsorption Assay” is an assay used to assess Cationic Monomer, Cationic Monomeric Unit or Cationic the capacity of a Surface for fibrinogen. In the assay, test Repeat Unit: unless otherwise indicated, a "cationic mono 45 samples are placed in a suitable sized container, which may be mer.” “cationic monomeric unit' or “cationic repeat unit is a a 96-well manifold, microcentrifuge tube, or other container. monomer or a monomeric or repeat unit (the terms "mono The Volumes in the following are appropriate for a deep meric unit' and “repeat unit' being used interchangeably) 96-well plate, but may be scaled to properly cover a device bearing a cation or other cationic species, e.g., a moiety being tested. The samples are sterilized with 70% ethanol capable of having a positive charge upon addition of an elec 50 Solution for thirty minutes and the test groups run with an in trophile (e.g., a proton (H+) or an alkyl cation, for example in per run of 3-4. The sample container is blocked with 20 a pH dependent manner) or removal of a protecting group or mg/mL Bovine Serum Albumin (BSA) in 1xEBS for 1 hour at a nucleophile). 4°C., followed by three rinses with 1xPBS before samples Cationic species or Cationic Moiety: unless otherwise indi are added. The sample is exposed to a solution containing 70 cated, a “Cationic species” or a "Cationic Moiety” is a group, 55 ug/mL unlabeled human fibrinogen, 1.4 g/mL I-125 radio residue or molecule that is present in a positively charged or labeled human fibrinogen, 35-55 ug/mL BSA in water, non-charged State, but in the non charged State is capable of optionally tri-sodium citrate, and optionally sodium chloride. becoming positively charged, e.g., upon addition of an elec The BSA is a common agent co-lyophilized with the radio trophile (e.g., a proton (H+), for example in a pH dependent labeled fibrinogen. Optionally, the BSA and radiolabeled manner) or removal of a protecting group or a nucleophile. In 60 fibrinogen may have been dissolved from a lyophilized form certain instances, the group, residue or molecule is perma that contains tri-sodium citrate and Sodium chloride. The nently charged, e.g., comprises a quaternary nitrogen atom. samples are incubated for one hour at 37° C. on an orbital Coating: unless otherwise indicated, "coating refers to shaker at 150 RPM. The test solution is then removed and four any temporary, semi-permanent or permanent layer, or layers, 1-minute rinses with a 10 mM NaI and one 1-minute rinse treating or covering a surface. The coating may be a chemical 65 with 1xPBS is performed. The samples are loaded into a modification of the underlying substrate or may involve the gamma counter. The counter measures the radioactivity in addition of new materials to the surface of the substrate. It I-125 counts per minute for each sample and this data is used US 9,096,703 B2 13 14 to calculate the absolute fibrinogen adsorption or a percent Graft-from method: unless otherwise indicated, the term reduction of the non-fouling polymer layer samples Versus a 'graft-from, as used herein in connection with a method for reference Substrate, that is, the same or an otherwise func the modification of a material with a polymer, shall mean the tionally equivalent Substrate lacking the non-fouling polymer in situ polymerization and growth of a polymer at the Surface layer. The percent reduction is equal to: (1-non-fouling 5 of, or within a material. sample CPM/Average CPM of the reference substrate) Graft-from polymer: unless otherwise indicated, the term * 100%. 'graft-from polymer, as used herein, shall mean a polymer Global Average Dry Thickness: unless otherwise indi formed by a graft-from method. cated, “Global Average Dry Thickness.” as used herein in Graft-through method: unless otherwise indicated, the connection with a polymer layer, shall mean the mean calcu 10 term 'graft-through as used herein in connection with a lated by averaging the Local Average Dry Thickness of at method for the modification of a material with a polymer, least 3, and preferably at least 5, representative locations shall mean the in situ polymerization of monomers in the spaced approximately evenly across the portion of the article neighborhood of the material that may polymerize through carrying the polymer layer. For example, if a polymer layer is functional groups presented from the material Surface. For applied to the indwelling portion of a catheter, the represen 15 example, the material may have vinyl groups presented from tative locations are approximately evenly spaced across the the Surface through which polymerization occurs. indwelling portion of the catheter. It is preferred to measure Graft-through polymer: unless otherwise indicated, the the thickness at representative points across the longest term 'graft-through polymer, as used herein, shall mean a dimension of the portion of the article that is covered with the polymer formed by a graft-through method. polymer layer. The standard deviation of the Global Average Graft-to method: unless otherwise indicated, the term Dry Thickness is found by calculating the standard deviation 'graft-to.” as used herein in connection with a method for the of the Local Average Dry Thickness across at least 5, and modification of a material with a polymer shall mean the preferably at least 10, representative locations spaced modification of the surface of a material with a presynthe approximately evenly across the portion of the article carry 25 sized polymer ing the polymer layer. Graft-to polymer: unless otherwise indicated, the term Global Average Humidified Thickness: unless otherwise 'graft-topolymer, as used herein, shall mean a grafted poly indicated, “Global Average Humidified Thickness, as used merformed by a graft-to method. herein in connection with a polymer layer, shall mean the Heteroalkyl: unless otherwise indicated, the term “het mean calculated by averaging the Local Average Humidified 30 Thickness of at least 3, and preferably at least 5, representa eroalkyl means an alkyl group wherein at least one of the tive locations spaced approximately evenly across the portion backbone carbon atoms is replaced with a heteroatom. of the article carrying the polymer layer. For example, if a Heteroaryl: unless otherwise indicated, the term “het polymer layer is applied to the indwelling portion of a cath eroaryl' means an aryl group wherein at least one of the ring eter, the representative locations are approximately evenly 35 members is a heteroatom, and preferably 5 or 6 atoms in each spaced across the indwelling portion of the catheter. It is ring. The heteroaromatic group preferably has 1 or 2 oxygen preferred to measure the thickness at representative points atoms, 1 or 2 sulfur atoms, and/or 1 to 4 nitrogenatoms in the across the longest dimension of the portion of the article that ring, and may be bonded to the remainder of the molecule is covered with the polymer layer. The standard deviation of through a carbon or heteroatom. Exemplary heteroaromatics the Global Average Humidified Thickness is found by calcu 40 include furyl, thienyl, pyridyl, oxazolyl pyrrolyl, indolyl, lating the standard deviation of the Local Average Humidified quinolinyl, or isoquinolinyl and the like. Exemplary Substitu Thickness across at least 5, and preferably at least 10, repre ents include one or more of the following groups: hydrocar sentative locations spaced approximately evenly across the byl, substituted hydrocarbyl, keto (i.e., =O), hydroxy, pro portion of the article carrying the polymer layer. tected hydroxy, acyl, acyloxy, alkoxy, alkenoxy, alkynoxy, Global Average R. Surface Roughness: unless otherwise 45 aryloxy, halogen, amido, amino, nitro, cyano, thiol, ketals, indicated, “Global Average R. Surface Roughness, as used acetals, esters and ethers. herein in connection with a polymer layer, shall mean the Heteroatom: unless otherwise indicated, the term "heteroa mean calculated by averaging the R. Surface roughness of at tom' means an atom other than hydrogen or carbon, such as least 5, and preferably at least 10, representative locations a chlorine, iodine, bromine, oxygen, Sulfur, nitrogen, phos spaced approximately evenly across the portion of the article 50 carrying the polymer layer. For example, if a polymer layer is phorus, boron, arsenic, selenium or silicon atom. applied to the indwelling portion of a catheter, the represen Heterocyclo: unless otherwise indicated, the terms “het tative locations are approximately evenly spaced across the erocyclo' and "heterocyclic” as used herein alone or as part of indwelling portion of the catheter. It is preferred to measure another group denote optionally Substituted, fully saturated the thickness at representative points across the longest 55 or unsaturated, monocyclic or bicyclic, aromatic or nonaro dimension of the portion of the article that is covered with the matic groups having at least one heteroatom in at least one polymer layer. The standard deviation of the Global Average ring, and preferably 5 or 6 atoms in each ring. The heterocyclo R. Surface Rroughness is found by calculating the standard group preferably has 1 or 2 oxygenatoms, 1 or 2 sulfur atoms, deviation of the Local Average R. Surface Roughness and/or 1 to 4 nitrogenatoms in the ring, and may be bonded to across at least 5, and preferably at least 10, representative 60 the remainder of the molecule through a carbon or heteroa locations spaced approximately evenly across the portion of tom. Exemplary heterocyclo include heteroaromatics such as the article carrying the polymer layer. furyl, thienyl, pyridyl, oxazolyl pyrrolyl, indolyl, quinolinyl, Graft: unless otherwise indicated, the term 'graft, as used or isoquinolinyl and the like. Exemplary Substituents include herein in connection with a polymer, means the modification one or more of the following groups: hydrocarbyl, Substituted of the surface of a material with a polymer by a 'graft-from'. 65 hydrocarbyl, keto, hydroxy, protected hydroxy, acyl, acyloxy, 'graft-through', or a 'graft-to” approach, or a combination alkoxy, alkenoxy, alkynoxy, aryloxy, halogen, amido, amino, thereof to form a grafted polymer. nitro, cyano, thiol, ketals, acetals, esters and ethers. US 9,096,703 B2 15 16 Heterohydrocarbyl: unless otherwise indicated, the term representative locations spaced approximately evenly across "heterohydrocarbyl means a hydrocarbyl group wherein at a cross section of article that spans approximately 10-40 least one of the chain carbon atoms is replaced with a het micrometers. erOatOm. Local Average Humidified Thickness: unless otherwise Humidified Thickness: unless otherwise indicated, indicated, “Local Average Humidified Thickness” is the "humidified thickness, as used herein in connection with a mean Humidified Thickness calculated by averaging polymer layer, shall mean the thickness of the polymer layer Humidified Thickness measurements of at least 3, and pref using an environmental scanning electron microscope erably at least 5, representative locations spaced approxi (ESEM and approximately 26% relative humidity). To mea mately evenly across a cross section of the article that spans 10 approximately 10-40 micrometers. The standard deviation of sure humidified thickness, the sample is freeze fractured for the Local Average Humidified Thickness may be determined imaging by being Submerged in liquid nitrogen then cracked by calculating the standard deviation of the Humidified with an ultra microtome blade. The freeze fracturing should Thickness across of at least 5, and preferably at least 10, break the article at a plane orthogonal to the polymer modified representative locations spaced approximately evenly across surface in order to measure the thickness of the polymer layer 15 a cross section of article that spans approximately 10-40 normal to the Substrate. After fracturing, the samples are micrometers. soaked in water for at least one hour and then Submerged in Membrane-Targeting Antimicrobial Agent: unless other liquid nitrogen and fixed to a cold stage at -8°C. to -12°C. wise indicated, “membrane-targeting antimicrobial agent' The samples are then imaged using a VPSE detector at the refers to any antimicrobial agent that retains its bactericidal or highest resolvable humidity (approximately 26% or 81 Pa) bacteriostatic activity when immobilized on a substrate and under a Scanning Electron Microscope (SEM) with an Envi can therefore be used to create an immobilized antimicrobial ronmental Scanning Electron Microscope (E-SEM). Exem Surface. In one embodiment, the membrane-targeting antimi plary microtome blades include the Leica Ultracut UCT crobial agent is an antimicrobial peptide, and in another Ultramicrotome, exemplary SEMs include the Supra55VP embodiment it is a quaternary ammonium compound or poly FESEM, Zeiss, and exemplary E-SEMs include the Zeiss 25 C. EVO 55. Non-Degradable: unless otherwise indicated, “non-de Hydrocarbon or Hydrocarbyl: unless otherwise indicated, gradable' refers to material compositions that do not react the terms “hydrocarbon and “hydrocarbyl as used herein significantly within a biological environment either hydro describe organic compounds or radicals consisting exclu lytically, reductively, enzymatically or oxidatively to cleave sively of the elements carbon and hydrogen. These moieties 30 into Smaller or simpler components. include alkyl, alkenyl, alkynyl, and aryl moieties. These moi Non-Fouling Composition/Non-Fouling Material/Non eties also include alkyl, alkenyl, alkynyl, and aryl moieties Fouling Polymer/Non-Fouling Polymer Layer: unless other Substituted with otheraliphatic or cyclic hydrocarbon groups, wise indicated, a “non-fouling composition' or “non-fouling Such as alkaryl, alkenaryl and alkynary1. Unless otherwise material' or “non-fouling polymer or “Non-fouling polymer indicated, these moieties preferably comprise 1 to 20 carbon 35 layer” as used interchangeably herein, is a composition that atOms provides or increases the protein resistance of a Surface of an Hydrophilic: unless otherwise indicated, “hydrophilic' article to which the composition is attached. For example, refers to solvents, molecules, compounds, polymers, mix when attached to a Substrate such a composition may resist tures, materials, or functional groups which have an affinity the adhesion of proteins, including blood proteins, plasma, for water. Such materials typically include one or more 40 cells, tissue and/or microbes to the substrate relative to the hydrophilic functional groups, such as hydroxyl, Zwitteri amount of adhesion to a reference Substrate, that is, the same onic, carboxy, amino, amide, phosphate, Sulfonyl, hydrogen or an otherwise functionally equivalent Substrate lacking the bond forming, and/or ether groups. composition. Preferably, a substrate surface will be substan Hydrophobic: unless otherwise indicated, “hydrophobic' tially non-fouling in the presence of human blood. Preferably refers to solvents, molecules, compounds, polymers, mix 45 the amount of adhesion will be decreased 20%, 30%, 40%, tures, materials, or functional groups that are repelled by 50%. 60%, 70%, 80%, or more, for example, 85%, 90%, water. Such materials typically contain non-polar functional 95%, 99%, 99.5%, 99.9%, or more, relative to the reference groups. substrate. One particularly preferred measure of the non Immobilization/Immobilized: unless otherwise indicated, fouling character or protein resistance of a surface is the “immobilization' or “immobilized’ refers to a material or 50 amount of fibrinogen adsorbed in a Fibrinogen Adsorption bioactive agent that is covalently or non-covalently attached Assay as described herein. Preferably, the amount of directly or indirectly to a substrate. “Co-immobilization' adsorbed fibrinogen using the Fibrinogen Adsorption Assay refers to immobilization of two or more agents. described herein is <125 ng/cm, <90 ng/cm, <70 ng/cm, Initiator: unless otherwise indicated, “initiator” refers to a <50 ng/cm, <30 ng/cm, <20 ng/cm, <15 ng/cm, <12 Substance or a combination of Substances that can produce a 55 ng/cm, <10 ng/cm, <8 ng/cm, <6 ng/cm, <4 ng/cm, <2 radical or other species under relatively mild conditions and ng/cm, <1 ng/cm, <0.5 ng/cm, or <0.25 ng/cm. promote polymerization reactions. For example, redox pairs Non-Naturally Occurring Amino Acid: unless otherwise as described elsewhere herein may be an initiator. indicated, “non-naturally occurring amino acid refers to any Local Average Dry Thickness: unless otherwise indicated, amino acid that is not found in nature. Non-natural amino “Local Average Dry Thickness” is the mean Dry Thickness 60 acids include any D-amino acids, amino acids with side calculated by averaging Dry Thickness measurements of at chains that are not found in nature, and peptidomimetics. least 3, and preferably at least 5, representative locations Examples of peptidomimetics include, but are not limited to, spaced approximately evenly across a cross section of the b-peptides, g-peptides, and d-peptides; oligomers having article that spans approximately 10-40 micrometers. The backbones which can adopt helical or sheet conformations, standard deviation of the Local Average Dry Thickness is 65 Such as compounds having backbones utilizing bipyridine determined by calculating the standard deviation of the Dry segments, compounds having backbones utilizing Solvopho Thickness across at least 5, and more preferably at least 10, bic interactions, compounds having backbones utilizing side US 9,096,703 B2 17 18 chain interactions, compounds having backbones utilizing meter. For example, the sample Surface roughness can be hydrogen bonding interactions, and compounds having back measured by a Tencor P-16+ profilometer with a 60 degree, 2 bones utilizing metal coordination. All of the amino acids in um diamond tip stylus. Preferably, an 800 um Scan length is the human body, except glycine, exist as the D and L forms. chosen with 20 Lum/second scan rate, 50 Hz scan frequency, Nearly all of the amino acids occurring in nature are the and 2 ug loading force. At least three different sites are mea L-forms. D-forms of the amino acids are not found in the Sured for the same sample, and the Surface roughness is aver proteins of higher organisms, but are present in some lower aged from at least three samples. Alternatively, the R. Sur forms of life, such as in the cell walls of bacteria. They also face roughness can be measured preferably by non-contact are found in some antibiotics, among them, streptomycin, methods, including using optical profilometers. For example, actinomycin, bacitracin, and tetracycline. These antibiotics 10 the sample surface roughness is measured by a optical pro can kill bacterial cells by interfering with the formation of filometer (Zeta Z20 or Olympus Lext OLS4000). Preferably proteins necessary for viability and reproduction. Non-natu a 3-D image is taken by the optical profilometer under a 50x rally occurring amino acids also include residues, which have objective lens, and the sample's Surface roughness is then side chains that resist non-specific protein adsorption, which measured along at least three different lines cross the image. may be designed to enhance the presentation of the antimi 15 At least three different spots are measured and the surface crobial peptide in biological fluids, and/or polymerizable side roughness is averaged from at least three samples. In a pre chains, which enable the synthesis of polymer brushes using ferred example an Olympus LEXT OLS4000 3D Laser Mea the non-natural amino acid residues within the peptides as Suring Microscope is employed for roughness measurements monomeric units. and 3D imaging. A LEXT microscope utilizes low wave Polymer: unless otherwise indicated, “polymer includes length optical technology with a 408 nm laser in combination natural and synthetic, homopolymers and copolymers com with confocal scanning can be used for the measurement. prising multiple repeat units and, unless otherwise indicated, Samples to be measured are mounted on a glass slide by may be linear, branched, or dendritic. Examples of copoly double-sided tape. Digital 3-D images are taken with the mers include, but are not limited to, random copolymers and Olympus LEXT OLS4000 laser confocal microscope block copolymers, Smart polymers, temperature responsive 25 (“LEXT) under an Olympus MPLAPON 50x objective lens. (e.g., NIPAM), and pH responsive (e.g., pyridyl based) poly The digital images taken in this way have a 256x256 um field CS. area. The Z-direction repeatability for this LEXT machine Polypeptide/Peptide/Oligopeptide: unless otherwise indi has been certified by Olympus to be less than 0.012 um. To cated, “polypeptide.” “peptide.” and "oligopeptide' encom measure the roughness, at least three images have been taken pass organic compounds composed of amino acids, whether 30 from each sample and the R, roughness is calculated by natural, synthetic or mixtures thereof, that are linked together using a 9 um cut-off length. chemically by peptide bonds. Peptides typically contain 3 or Solvent Extractable Polymerization Initiator: unless other more amino acids, preferably more than 9 and less than 150, wise indicated, “Solvent Extractable Polymerization Initia more preferably less than 100, and most preferably between 9 tor” refers to any compound capable of starting radical poly and 51 amino acids. The polypeptides can be “exogenous.” or 35 merization that has been incorporated within the article, "heterologous, i.e., production of peptides within an organ wherein either the initiator or its degradation products may be ism or cell that are not native to that organism or cell. Such as extracted from the article using a Suitable solvent. In general, human polypeptide produced by a bacterial cell. Exogenous extractions can use nonpolar or polar solvents. For example, also refers to Substances that are not native to the cells and are extraction solvents such as water, acetone or ethanol; and/or added to the cells, as compared to endogenous materials, 40 other extraction solvents in which the solubility of the initia which are produced by the cells. The peptide bond involves a tor and/or its degradation products is at least 1 mg/L. The single covalent link between the carboxyl group (oxygen extraction should be carried out for a sufficient time such that bearing carbon) of one amino acid and the amino nitrogen of the change in concentration of the extract is not increasing a second amino acid. Small peptides with fewer than about ten more than 5% per hour. Alternatively, extraction until the constituent amino acids are typically called oligopeptides, 45 amount of extracted materialina Subsequent extraction is less and peptides with more than ten amino acids are termed than 10% of that detected in the initial extraction, or until polypeptides. Compounds with molecular weights of more there is no analytically significant increase in the cumulative than 10,000 Daltons (50-100 amino acids) are usually termed extracted material levels detected. Extraction conditions proteins. include: 37° C. for 72 h; 50° C. for 72 h; 70° C. for 24 h; 121° Quaternary Nitrogen: unless otherwise indicated, “quater 50 C. for 1 h. Extraction ratio includes 6 cm/mL surface area/ nary nitrogen as used herein, refers to a nitrogenatom that is Volume and/or 0.2 g sample/mL. In some instances, complete a member of a quaternary ammonium cation. dissolution of the substrate may be appropriate. Materials R. Surface Roughness: unless otherwise indicated, “R, shall be cut into small pieces before extraction to enhance Surface Roughness' refers to root mean squared roughness of Submersion in the extract media, for example, for polymeric a Surface, which measures the Vertical deviations of a real 55 substrates approximately 10 mmx50mm or 5mmx25mm are surface from its ideal form. The roughness refers to surface appropriate. The instrumentation used includes high-perfor micro-roughness which may be different than measurements mance liquid chromatography-photo-diode array detection of large scale surface variations. Preferably, this may be mea mass spectrometry (HPLC-PDA-MS) for organics analysis: sured using atomic force microscopy (MFP-3D, Aslyum) gas chromatography-mass spectrometry (GC-MS) for organ across a field of approximately 1-30 um by 1-30 um, prefer 60 ics analysis; inductively coupled plasma-optical emission ably 20 um by 20 Lum. The sample is washed with purified spectroscopy or mass spectrometry (ICP-OES or ICP-MS) water to remove surface salts and then air dried. Standard for metals analysis; and sometimes ion chromatography (IC) silicon cantilever (Olympus AC160TS, spring constant 42 for inorganics and ion analysis. Sometimes more advanced N/m) is employed for the measurement with an AC/Tapping MS detectors such as time-of-flight (TOF) are used to obtain mode. The R. Surface roughness is calculated by the Soft 65 accurate mass information. Hexane and alcohol extractions ware (IGOR Pro) attached with the AFM machine. Alterna are analyzed by GC-MS. Water and alcohol extractions are tively the roughness can be measured using a stylus profilo analyzed by HPLC. The initiator or its degradation products US 9,096,703 B2 19 20 may be quantified and/or detected in the substrate or grafted the composition does not lyse 50%, preferably 20%, more polymer by the previously described methods. These include preferably 10%, even more preferably 5%, most preferably FTIR-ATR, electron spectroscopy for chemical analysis 1%, of human red blood cells when the following assay is (ESCA, also called X-ray photoelectron spectroscopy, XPS), applied: a stock of 10% washed pooled red blood cells (Rock Secondary Ion Mass Spectrometry (SIMS), and surface-en land Immunochemicals Inc, Gilbertsville, Pa.) is diluted to hanced Raman spectroscopy (SERS). For example, peroxide 0.25% with a hemolysis buffer of 150 mM. NaCl and 10 mM may be detected spectrophotometrically using any of the Tris at pH 7.0. A 0.5 cm antimicrobial sample is incubated following three methods: the iodide method (oxidation of with 0.75 mL of 0.25% red blood cell suspension for 1 hour at sodium iodide by peroxides in the presence of ferric chlo 37°C. The solid sample is removed and cells are spun down ride), the DPPH method (treatment with 1,1-diphenyl-2-pic 10 rylhydrazyl, a radical Scavenger, to decompose the peroX at 6000 g, the supernatant is removed, and the OD414 mea ides), or the peroxidase method (reduction with glutathione, sured on a spectrophotometer. Total hemolysis is defined by catalyzed by glutathione peroxidase, followed by measuring diluting 10% of washed pooled red blood cells to 0.25% in the coupled oxidation of NADPH in the presence of glu sterile deionized (DI) water and incubating for 1 hour at 37° tathione reductase). See, for example, Fujimoto et al., Journal 15 C., and 0% hemolysis is defined using a suspension of 0.25% of Polymer Science Part A: Polymer Chemistry, Vol. 31, red blood cells in hemolysis buffer without a solid sample. 1035-1043 (1993). Substantially Non-Toxic: unless otherwise indicated, Stable: unless otherwise indicated, “stable, as used herein “Substantially non-toxic' means a Surface that is Substantially in reference to a material, means that the material retains hemocompatible and Substantially non-cytotoxic. functionality over extended periods of time. In one embodi Substituted/Optionally Substituted: unless otherwise indi ment, the referenced material retains at least 90% of a refer cated, the term “substituted” and “optionally substituted enced activity (or property) for at least 30 days at 37°C. in at means that the referenced group is or may be substituted with least one of phosphate buffered saline containing protein, one or more additional Suitable group(s), which may be indi media, or serum, or in vivo. In one embodiment, the reference vidually and independently selected, for example, from material retains at least 80% of a referenced activity (or 25 acetals, acyl, acyloxy, alkenoxy, alkoxy, alkylthio, alkynoxy, property) for at least 90 days at 37° C. in at least one of amido, amino, aryl, aryloxy, arylthio, azido, carbonyl, car phosphate buffered saline containing protein, media, or boxamido, carboxyl, cyano, esters, ethers, hydrocarbyl, Sub serum, or in vivo. In one embodiment, the referenced material stituted hydrocarbyl, heterohydrocarbyl, substituted hetero retains at least 90% of the referenced activity (or property) for hydroalkyl, cycloalkyl, halogen, heteroalicyclic, heteroaryl, at least 30 days at 37° C. and at least 80% of the referenced 30 hydroxy, isocyanato, isothiocyanato, ketals, keto, mercapto, activity (or property) for at least 90 days at 37°C. The refer enced activity or property may include surface contact angle, nitro, perhaloalkyl, silyl, Sulfamoyl, Sulfate, sulfhydryl, Sul non-fouling, anti-thrombogenic, and/or antimicrobial activ fonamido, Sulfonate, Sulfonyl, Sulfoxido, thiocarbonyl, thio ity. cyanato, thiol, and/or the protected derivatives thereof. It will be understood that “substitution' or “substituted” includes Static Contact Angle: unless otherwise indicated, “Static 35 Contact Angle' is the angle at which a water/vapor interface the implicit proviso that such Substitution is in accordance meets a Substrate Surface at or near equilibrium conditions. with permitted valence of the substituted atom and the sub The contact angle is measured by first soaking the samples stituent, and that the Substitution results in a stable com with pure ethanol for 5 minutes and washing with PBS three pound, e.g., which does not spontaneously undergo transfor times. The samples are then soaked within PBS (150 mM, pH 40 mation Such as by rearrangement, cyclization, elimination, 7.4) for 24 hours and washed three times with purified water. etc. Then the samples are dried under a flow of airfor 5 min before Substrate: unless otherwise indicated, “substrate” refers to testing. A drop of purified water (e.g., 1 LL) is deposited on the material from which a non-fouling polymer is grafted. the test Surface, the shape of the droplet is photographed by a Sulfoammonium: unless otherwise indicated, a "sulfoam microscope with a CCD camera using a video contact angle 45 monium moiety is a Zwitterionic moiety comprising Sulfate system (e.g., VCA 2000, AST Inc.), and the contact angle is and ammonium functionality and includes, for example, Sul then determined (using, for example, a VCA Optima XE). foammonium monomers, Sulfoammonium oligomers, Sul The size of the water droplet used to determine the contact foammonium polymers, Sulfoammonium repeat units, and angle may vary depending upon the Substrate type and com other Sulfoammonium-containing materials. Sulfobetaine position. For a 5 French device, for instance, an 0.1 uL drop of 50 monomers, oligomers, polymers, repeat units, and other Sul purified water may be used. Substantially Hemocompatible: unless otherwise indi fobetaine materials are exemplary Sulfoammonium moieties. cated, “substantially hemocompatible” means that the com Tether/Tethering Agent/Linker: unless otherwise indi position is Substantially non-hemolytic, in addition to being cated, “tether or “tethering agent' or “linker as used herein non-thrombogenic and non-immunogenic, as tested by 55 synonymously, refers to any molecule, or set of molecules, or appropriately selected assays for thrombosis, coagulation, polymer used to covalently or non-covalently immobilize one and complement activation as described in ISO 10993-4. or more non-fouling materials, one or more bioactive agents, Substantially Non-Cytotoxic: unless otherwise indicated, or combinations thereof on a material where the molecule “substantially non-cytotoxic' refers to a composition that remains as part of the final chemical composition. The tether does not Substantially change the metabolism, proliferation, 60 can be either linear or branched with one or more sites for or viability of mammalian cells that contact the surface of the immobilizing bioactive agents. The tether can be any length. composition. These may be quantified by the International However, in one embodiment, the tether is greater than 3 Standard ISO 10993-5 which defines three main tests to angstroms in length. The tether may be non-fouling, such as assess the cytotoxicity of materials including the extract test, a monomer, oligomer, or polymer or a non-fouling non-Zwit the direct contact test and the indirect contact test. 65 terionic material. The tether may be immobilized directly on Substantially Non-Hemolytic Surface: unless otherwise the substrate or on a polymer, either of which may be non indicated, “substantially non-hemolytic surface” means that fouling. US 9,096,703 B2 21 22 Undercoating Layer: unless otherwise indicated, “under of such materials; for example, manufacturing methods may coating layer” refers to any coating, or combination of coat affect the porosity of a material, its roughness (micro-rough ings, incorporated into a Substrate from which a non-fouling ness and macro-roughness), incorporation of foreign-body polymer is grafted. inclusions that project from the Surface of the material, and Zwitterion/Zwitterionic Material: unless otherwise indi similar Surface characteristics. Each of these, and other fac cated, “Zwitterion' or “Zwitterionic material' refers to a mac tors, may contribute to a materials resistance (or lack romolecule, material, or moiety possessing both cationic and thereof) to protein adsorption and/or cell/microorganism anionic groups. In most cases, these charged groups are bal adhesion. anced, resulting in a material with Zero net charge. Without being bound to any particular theory, it is presently Zwitterionic Polymers: unless otherwise indicated, “Zwit 10 terionic polymers' may be homopolymers or copolymers and believed that the graft-from polymerization methods of the include both polyampholytes (e.g., polymers with the present invention provide a surface modification, that is, a charged groups on different monomer units) and polybetaine non-fouling polymer layer, having a branched structure (polymers with the anionic and cationic groups on the same which disfavors protein adsorption and/or cell/microorgan monomer unit). Exemplary Zwitterionic polymers include 15 ism adhesion and which may, in addition, conceal or other alternating copolymers, statistical copolymers, random wise alter the sites in a substrate that favor the adhesion of copolymers and block copolymers of two, three or more cells, bacteria or other microorganisms. Thus, for example, and relative to the (unmodified) surface of the article, the OOCS. grafted polymer layer may cover or even, partially or com DETAILED DESCRIPTION OF THE INVENTION pletely fill, scratches, pinholes, voids or other defects in the surface of the article that could potentially otherwise serve as Among the various aspects of the present invention may be a site for a performance failure. By way of further example, noted the provision of articles, such as medical devices, hav grafted polymer layers having a thickness that is at least as ing a non-fouling grafted polymer layer. In general, therefore, great as the Surface roughness of the (unmodified) Surface of the article comprises a Substrate and a polymeric material 25 the article, that are relatively uniform, that are sufficiently grafted from the substrate. Surprisingly, it has been found that dense, and/or are significantly hydrophilic can significantly non-fouling grafted polymer layers can be provided by incor increase a materials resistance to protein adsorption and/or porating one or more polymerization initiator(s) into the Sub cell/microorganism adhesion. strate, for example, by imbibing the substrate with the initia In one aspect of the present invention, a non-fouling layer tor(s) or depositing a layer onto the Substrate that comprises 30 is applied onto only a portion or portions of a Substrate or the initiator(s), and grafting a polymer from the Substrate. In object, including in a 2 or 3-dimensional pattern or patterns at a particularly preferred embodiment, the polymeric material discrete locations on a substrate or object. In some embodi is grafted from the Substrate in a polymerization mixture ments the non-fouling layer is applied onto a Substrate or comprising monomer and a solvent system wherein the Sub object in Such a way as to have discrete and/or blended geo strate is not significantly Swelled by the solvent system and 35 metrical features and/or designs at many scales ranging from the incorporated initiator has limited solubility in the solvent nanometers to micrometers to millimeters. Preferred embodi system. Stated differently, the initiator(s) incorporated into ments include controllably forming a discrete non-fouling the substrate have reversed phase properties compared to the feature(s) involves the selective masking or blocking of the solvent system especially in terms of hydrophilicity. Without desired portions of the substrate from imbibing and/or appli being bound to any particular theory, it is believed that this 40 cation of the initiator and/or from the graft polymerization. In method provides a relatively high local concentration of ini one embodiment a portion of the Substrate article is masked tiator(s) at or near the Substrate Surface/polymerization mix during initiator application. In one preferred embodiment a ture interface, and favors grafting from the Substrate and the portion of the Substrate article is masked during polymeriza grafted polymer to form a branched polymer. tion. The masking technique may be applied to any of the Regardless of the theory, the grafted polymers of the 45 Substrates described herein, including metals, ceramics, present invention comprise a relatively dense, branched and glasses, polymers, biological tissues, living or dead, woven hydrophilic structure that uniformly covers substrate surface and non-woven fibers, semi-metals such as silicon. defects and enhances performance. As a result, articles having Among the various aspects of the present invention is con a surface modified by the grafted polymers possess improved trollably placing a non-fouling polymer at discrete locations anti-fouling, and/or antithrombotic characteristics and, in 50 on a Substrate or object by several means. For locations, certain embodiments, improved antimicrobial characteris patterns, geometric features/designs greater than or equal to tics. millimeter scale controllably placing a non-fouling polymer Generally speaking, Small initiator molecules can be con may be accomplished by physically masking areas where the centrated at or near the Substrate Surface, where polymeriza non-fouling polymer will not form, for example, by using tion is initiated and propagated, more readily than larger 55 techniques such as applying tape, screens, resists, or other polymer molecules synthesized in solution. As a result, and as blocking materials that inhibit access of the polymerization compared to graft-to coatings, greater Surface densities can be solution to the substrate or object surface, therefore inhibiting achieved which, in turn, tends to improve non-fouling perfor polymer formation. For locations, patterns, geometric fea mance. Additionally, longer polymer chains and/or branched tures/designs less than millimeter scale controllably placing a non-fouling chains may further improve performance. 60 non-fouling polymer may be accomplished by physically Medical devices and other articles comprise any of a wide masking areas where the non-fouling polymer will not form range of materials. Certain of these materials, by virtue of using techniques such as photolithographic procedures (such their intrinsic characteristics, exhibit a greater resistance to as stereolithography, laser-chemical three-dimensional writ protein adsorption and cell/microorganism adhesion; for ing, and modular assembly), microcontact printing, or example, hydrophilic materials tend to exhibit less protein 65 microstamping of blocking materials that inhibit access of the adsorption than hydrophobic materials. In addition, methods polymerization solution to the Substrate or object surface, of manufacture can greatly affect the Surface characteristics therefore inhibiting polymer formation. In addition, masking US 9,096,703 B2 23 24 materials can be applied at any scale by means of digital a fibrinogen adsorption of less than 12 ng/cm in such an application methods such as spray jet, valve jet, and inkjet assay. In some embodiments, the modified Surface exhibits a printing methods. fibrinogen adsorption of less than 10 ng/cm in such an assay. In some embodiments, masking during polymerization In some embodiments, the modified surface exhibits a permits the ability to apply a mix of different polymers over fibrinogen adsorption of less than 8 ng/cm in such an assay. different areas of a substrate or object. By means of selec In some embodiments, the modified surface exhibits a tively removing portions or types of masking materials or fibrinogen adsorption of less than 6 ng/cm in such an assay. agents from discrete locations followed by Subsequent poly In some embodiments, the modified surface exhibits a merization steps with different monomers or monomer mix fibrinogen adsorption of less than 4 ng/cm in such an assay. tures a non-fouling Surface can be constructed consisting of 10 In some embodiments, the modified surface exhibits a any number of different polymers with similar or different fibrinogen adsorption of less than 2ng/cm in such an assay. non-fouling, dimensional, mechanical, physical, and/or In some embodiments, the modified surface exhibits a chemical properties. fibrinogen adsorption of less than 1 ng/cm in such an assay. One embodiment includes techniques that remove initiator In some embodiments, the modified surface exhibits a imbibed portions and/or non-fouling polymer portions from 15 fibrinogen adsorption of less than 0.5 ng/cm in such an assay. the substrate or object in a controlled fashion and thus create In some embodiments, the modified surface exhibits a locations, patterns, geometric features/designs of non-foul fibrinogen adsorption of less than 0.25 ng/cm in such an ing polymer at any scale, including laser ablation, abrasive assay. In one embodiment, the grafted polymer in each of the media stream/spray, or direct contact physical abrasion/ foregoing examples recited in this paragraph is a Zwitterionic Scraping. polymer. In one embodiment, the grafted polymer in each of The presence or absence of a non-fouling polymer in a the foregoing examples and embodiments recited in this para controlled design/pattern can be used to control and/or modu graph is a polymer containing carboxyammonium or Sul late the interaction, adsorption, desorption, of proteins and foammonium repeat units. In one embodiment, the grafted other biomolecules as well as control and/or modulate, inter polymer in each of the foregoing examples and embodiments action, adsorption, desorption, proliferation of cells (eukary 25 recited in this paragraph is a polymer containing Sulfobetaine otes, prokaryotes). Structures which may influence these pro or carboxybetaine repeat units. In one embodiment, the cesses including creating columns perpendicular to the article grafted polymer in each of the foregoing examples and Surface, channels along the Surface, or a number of other embodiments recited in this paragraph is a Zwitterionic poly geometrical patterns. The feature size or space between fea mer and the Zwitterionic polymer is grafted from a polyure tures may be Smaller, approximately the same size as, or 30 thane polymer. In one embodiment, the grafted polymer in larger than the protein or cell being influenced. Structures that each of the foregoing examples and embodiments recited in reduce adsorption may be synergistic with non-fouling poly this paragraph is a carboxyammonium or sulfoammonium mer Surface modifications to enhance non-fouling ability. polymer and the carboxyammonium or Sulfoammonium Independent of any theory, articles of the present invention polymer is grafted from a polyurethane polymer. In one having a modified Surface comprising a grafted polymer 35 embodiment, the grafted polymer in each of the foregoing exhibit low fibrinogen adsorption in a fibrinogen adsorption examples and embodiments recited in this paragraph is a assay. In general, the modified Surface exhibits a fibrinogen polymer containing Sulfobetaine or carboxybetaine repeat adsorption of less than 125 ng/cm in a fibrinogen adsorption units and the polymer containing Sulfobetaine or carboxybe assay in which samples are incubated for 60 minutes at 37°C. taine repeat units is grafted from a polyurethane polymer. in 70 ug/mL fibrinogen derived from human plasma, and the 40 Preferred embodiments also show reduction in thrombus amount of adsorbed fibrinogenis determined using a standard for Substrates having a grafted polymer layer of the present protocol, preferably by using radiolabeled fibrinogen. In one invention. For example, thrombus reduction of modified sub embodiment, the modified surface exhibits a fibrinogen strates, i.e., Substrates having a grafted polymer layer can be adsorption of less than 90 ng/cm in a fibrinogen adsorption assessed relative to a reference Substrate, i.e., the same or an assay in which samples are incubated for 60 minutes at 37°C. 45 otherwise functionally equivalent Substrate lacking the in 70 ug/mL fibrinogen derived from human plasma, and the grafted polymer layer, by exposing them to freshly harvested amount of adsorbed fibrinogenis determined using a standard bovine blood, heparinized, with radiolabeled platelets, in a protocol, preferably by using radiolabeled fibrinogen. In one flow loop for 2 hours. As an assessment of anti-thrombogenic embodiment, the modified surface exhibits a fibrinogen performance, samples are placed in an ex-vivo flow loop adsorption of less than 70 ng/cm in a fibrinogen adsorption 50 model of thrombosis. Anti-thrombogenic activity can be assay in which samples are incubated for 60 minutes at 37°C. evaluated using ex-vivo flow loop model of thrombosis. in 70 ug/mL fibrinogen derived from human plasma, and the Briefly, up to 10 liters of fresh blood are collected from a amount of adsorbed fibrinogenis determined using a standard single animal (bovine). This blood is heparinized to prevent protocol, preferably by using radiolabeled fibrinogen. In one coagulation, filtered to remove particulates, and autologous embodiment, the modified surface exhibits a fibrinogen 55 radio-labeled platelets are added. Within eight hours after adsorption of less than 50 ng/cm in a fibrinogen adsorption blood harvesting, coated and uncoated articles are placed in a assay in which samples are incubated for 60 minutes at 37°C. flow loop circuit, which pumps blood from a bath over the in 70 ug/mL fibrinogen derived from human plasma, and the article and thenbackinto the bath. A second internal flow loop amount of adsorbed fibrinogenis determined using a standard circuit can be established for Substrate containing a lumen by protocol, preferably by using radiolabeled fibrinogen. Pref 60 connecting the two ports of the Substrate through a 2nd peri erably, the modified surface exhibits a fibrinogen adsorption staltic pump. The size of tubing into which the article is of less than 30 ng/cm in such an assay. More preferably, in placed and speed of the blood flow may be adjusted based on certain embodiments the modified surface exhibits a fibrino the size of the article being tested. Preferably, when the gen adsorption of less than 20 ng/cm in such an assay. Still articles are 14-15.5 French dialysis catheters, they are placed more preferably, in certain embodiments the modified surface 65 in a flow loop circuit with tubing diameter of approximately exhibits a fibrinogen adsorption of less than 15 ng/cm in such 12.5-25.4 mm inner diameter. Blood is pumped in the outer an assay. In some embodiments, the modified Surface exhibits circuit at a rate of approximately 2.5 L/min, while blood in the US 9,096,703 B2 25 26 inner circuit is pumped at a rate of approximately ~200-400 37°C. In one preferred embodiment, antibiofilm activity of 2 ml/min. When the articles are 5 French PICC catheter shafts, log is achieved after 30 days storage in citrated human plasma they are placed in a flow loop circuit of approximately 6.4 mm at 37°C. Inafurther preferred embodiment, antibiofilm activ inner diameter and blood flow rate is approximately 200 ity of 2 log is achieved after 90 days storage in citrated human mL/min. The lumens may be locked with a solution, for plasma at 37° C. example saline, during evaluation. Alternatively, the distal tip Preferred embodiments show resistance to protein adsorp may be sealed, for example with epoxy, during evaluation. tion after extended exposure to PBS, which may indicate When the articles are 10 French rods, they are placed in a flow hydrolytic stability. In some embodiments, the modified sur loop circuit of approximately 6.4 mm inner diameter and face exhibits a fibrinogen adsorption of less than 125 ng/cm blood flow rate is approximately 200 ml/min. After 60-120 10 in a fibrinogen adsorption assay in which samples are incu minutes, the articles are removed, inspected visually for bated for 60 minutes at 37°C. in 70 g/mL fibrinogen derived thrombus formation, and adhered platelets are quantified from human plasma after 30 days exposure to PBS at 37°C. using a Gamma counter. For samples not containing alumen, In some embodiments, the modified surface exhibits a only an outer circuit may be used to measure thrombus on the fibrinogen adsorption of less than 90 ng/cm in a fibrinogen outside of the device. In this assay, preferred embodiments 15 adsorption assay in which samples are incubated for 60 min show at least an 80% reduction relative to reference substrate utes at 37°C. in 70 g/mL fibrinogen derived from human in adsorbed platelets and substantial visual reduction of plasma after 30 days exposure to PBS at 37° C. In some thrombus. For example, in certain embodiments there is at embodiments, the modified surface exhibits a fibrinogen least a 90% reduction in adsorbed platelets for modified sub adsorption of less than 70 ng/cm in a fibrinogen adsorption strates relative to reference substrates. Preferred embodi assay in which samples are incubated for 60 minutes at 37°C. ments show at least a 98% reduction in adsorbed platelets for in 70 ug/mL fibrinogen derived from human plasma after 30 modified substrates relative to reference substrates. Alterna days exposure to PBS at 37°C. In some embodiments, the tively, in a preferred embodiment, the thrombogenecity of a modified surface exhibits a fibrinogen adsorption of less than modified substrate is reduced relative to the non-modified 50 ng/cm in a fibrinogen adsorption assay in which samples substrate, after exposure to a 47% (w/v) sodium citrate solu 25 are incubated for 60 minutes at 37°C. in 70 ug/mL fibrinogen tion in DI water for greater than 3 days. Embodiments show a derived from human plasma after 30 days exposure to PBS at visual reduction of thrombus relative to for modified sub 37°C. In some embodiments, the modified surface exhibits a strates relative to reference substrates. Preferred embodi fibrinogen adsorption of less than 30 ng/cm in a fibrinogen ments show at least an 80% reduction of a modified substrate adsorption assay in which samples are incubated for 60 min relative to a reference substrate in adsorbed platelets and 30 utes at 37°C. in 70 g/mL fibrinogen derived from human substantial visual reduction of thrombus. Preferred embodi plasma after 30 days exposure to PBS at 37° C. In some ments show at least a 90% reduction in adsorbed platelets for embodiments, the modified surface exhibits a fibrinogen modified substrates relative to reference substrates. Preferred adsorption of less than 20 ng/cm in a fibrinogen adsorption embodiments show at least a 98% reduction in adsorbed assay in which samples are incubated for 60 minutes at 37°C. platelets for modified substrates relative to reference sub 35 in 70 ug/mL fibrinogen derived from human plasma after 30 strates. Alternatively, the thrombogenecity of preferred days exposure to PBS at 37°C. In some embodiments, the embodiments are reduced relative to the non-modified sub modified surface exhibits a fibrinogen adsorption of less than strate after exposure to animal serum and/or plasma. For 15 ng/cm in a fibrinogen adsorption assay in which samples example, the thrombogenecity of preferred embodiments are are incubated for 60 minutes at 37°C. in 70 ug/mL fibrinogen reduced after 55 day exposure to citrated human plasma at 37° 40 derived from human plasma after 30 days exposure to PBS at C. for modified substrates relative to reference substrates. 37°C. In some embodiments, the modified surface exhibits a Embodiments show a visual reduction of thrombus for modi fibrinogen adsorption of less than 12 ng/cm in a fibrinogen fied substrates relative to reference substrates. Preferred adsorption assay in which samples are incubated for 60 min embodiments show at least an 80% reduction for modified utes at 37°C. in 70 g/mL fibrinogen derived from human substrates relative to reference substrates in adsorbed plate 45 plasma after 30 days exposure to PBS at 37° C. In some lets and substantial visual reduction of thrombus. Preferred embodiments, the modified surface exhibits a fibrinogen embodiments show at least a 90% reduction in adsorbed adsorption of less than 10 ng/cm in a fibrinogen adsorption platelets for modified substrates relative to reference sub assay in which samples are incubated for 60 minutes at 37°C. strates. Preferred embodiments show at least a 98% reduction in 70 ug/mL fibrinogen derived from human plasma after 30 in adsorbed platelets for modified substrates relative to refer 50 days exposure to PBS at 37°C. In some embodiments, the ence Substrates. modified surface exhibits a fibrinogen adsorption of less than Preferred embodiments show antibiofilm activity for 8 ng/cm in a fibrinogen adsorption assay in which samples modified Substrates of at least 0.5 log, 1 log, 1.5 log, 2 log, 2.5 are incubated for 60 minutes at 37°C. in 70 ug/mL fibrinogen log, 3 log, or 4 log. More preferred embodiments have anti derived from human plasma after 30 days exposure to PBS at biofilm activity after extended exposures to PBS, serum, or 55 37°C. In some embodiments, the modified surface exhibits a plasma products. In one preferred embodiment, antibiofilm fibrinogen adsorption of less than 6 ng/cm in a fibrinogen activity of 1 logis achieved after 30 days storage in PBS at 37° adsorption assay in which samples are incubated for 60 min C. In a further preferred embodiment, antibiofilm activity of utes at 37°C. in 70 g/mL fibrinogen derived from human 1 log is achieved after 90 days storage in PBS at 37°C. In one plasma after 30 days exposure to PBS at 37° C. In some preferred embodiment, antibiofilm activity of 2 log is 60 embodiments, the modified surface exhibits a fibrinogen achieved after 30 days storage in PBS at 37° C. In a further adsorption of less than 4 ng/cm in a fibrinogen adsorption preferred embodiment, antibiofilm activity of 2 log is assay in which samples are incubated for 60 minutes at 37°C. achieved after 90 days storage in PBS at 37°C. In one pre in 70 ug/mL fibrinogen derived from human plasma after 30 ferred embodiment, antibiofilm activity of 1 log is achieved days exposure to PBS at 37°C. In some embodiments, the after 30 days storage in citrated human plasma at 37°C. In a 65 modified surface exhibits a fibrinogen adsorption of less than further preferred embodiment, antibiofilm activity of 1 log is 2ng/cm in a fibrinogen adsorption assay in which samples achieved after 90 days storage in citrated human plasma at are incubated for 60 minutes at 37°C. in 70 ug/mL fibrinogen US 9,096,703 B2 27 28 derived from human plasma after 30 days exposure to PBS at adsorption assay in which samples are incubated for 60 min 37°C. In some embodiments, the modified surface exhibits a utes at 37°C. in 70 g/mL fibrinogen derived from human fibrinogen adsorption of less than 1 ng/cm in a fibrinogen plasma after 90 days exposure to PBS at 37° C. In some adsorption assay in which samples are incubated for 60 min embodiments, the modified surface exhibits a fibrinogen utes at 37°C. in 70 ug/mL fibrinogen derived from human adsorption of less than 4 ng/cm in a fibrinogen adsorption plasma after 30 days exposure to PBS at 37° C. In some assay in which samples are incubated for 60 minutes at 37°C. embodiments, the modified surface exhibits a fibrinogen in 70 ug/mL fibrinogen derived from human plasma after 90 adsorption of less than 0.5 ng/cm in a fibrinogen adsorption days exposure to PBS at 37°C. In some embodiments, the assay in which samples are incubated for 60 minutes at 37°C. modified surface exhibits a fibrinogen adsorption of less than in 70 g/mL fibrinogen derived from human plasma after 30 10 2 ng/cm in a fibrinogen adsorption assay in which samples days exposure to PBS at 37° C. In some embodiments, the are incubated for 60 minutes at 37°C. in 70 ug/mL fibrinogen modified surface exhibits a fibrinogen adsorption of less than derived from human plasma after 90 days exposure to PBS at 0.25 ng/cm in a fibrinogen adsorption assay in which 37°C. In some embodiments, the modified surface exhibits a samples are incubated for 60 minutes at 37°C. in 70 g/mL fibrinogen adsorption of less than 1 ng/cm in a fibrinogen fibrinogen derived from human plasma after 30 days expo 15 adsorption assay in which samples are incubated for 60 min Sure to PBS at 37° C. utes at 37°C. in 70 g/mL fibrinogen derived from human Preferred embodiments show resistance to protein adsorp plasma after 90 days exposure to PBS at 37° C. In some tion after extended exposure to PBS, which may indicate embodiments, the modified surface exhibits a fibrinogen hydrolytic stability. In some embodiments, the modified sur adsorption of less than 0.5 ng/cm in a fibrinogen adsorption face exhibits a fibrinogen adsorption of less than 125 ng/cm assay in which samples are incubated for 60 minutes at 37°C. in a fibrinogen adsorption assay in which samples are incu in 70 ug/mL fibrinogen derived from human plasma after 90 bated for 60 minutes at 37°C. in 70 g/mL fibrinogen derived days exposure to PBS at 37°C. In some embodiments, the from human plasma after 90 days exposure to PBS at 37°C. modified surface exhibits a fibrinogen adsorption of less than In some embodiments, the modified surface exhibits a 0.25 ng/cm in a fibrinogen adsorption assay in which fibrinogen adsorption of less than 90 ng/cm in a fibrinogen 25 samples are incubated for 60 minutes at 37°C. in 70 ug/mL adsorption assay in which samples are incubated for 60 min fibrinogen derived from human plasma after 90 days expo utes at 37°C. in 70 ug/mL fibrinogen derived from human Sure to PBS at 37° C. plasma after 90 days exposure to PBS at 37° C. In some In general, the surface of the substrate may be modified embodiments, the modified surface exhibits a fibrinogen with any of a range of non-fouling polymeric materials. For adsorption of less than 70 ng/cm in a fibrinogen adsorption 30 example, the non-fouling polymeric material may be a assay in which samples are incubated for 60 minutes at 37°C. homopolymer or a copolymer. If a copolymer, the non-foul in 70 g/mL fibrinogen derived from human plasma after 90 ing polymeric material may be an alternating copolymer (e.g., days exposure to PBS at 37° C. In some embodiments, the AB . . . . a periodic copolymer (e.g., AB, ... wherein n modified surface exhibits a fibrinogen adsorption of less than and m are different), a statistical copolymer (a copolymer in 50 ng/cm in a fibrinogen adsorption assay in which samples 35 which monomers are arranged according to a known statisti are incubated for 60 minutes at 37°C. in 70 ug/mL fibrinogen cal rule), a random copolymer, or a block copolymer in which derived from human plasma after 90 days exposure to PBS at each of the blocks is independently a homopolymer or an 37°C. In some embodiments, the modified surface exhibits a alternating, periodic, statistical or random copolymer. Fur fibrinogen adsorption of less than 30 ng/cm in a fibrinogen thermore, when the non-fouling polymeric material is a adsorption assay in which samples are incubated for 60 min 40 copolymer it may be diblock, a triblock or other polyblock utes at 37°C. in 70 ug/mL fibrinogen derived from human copolymer. For example, in one preferred embodiment, the plasma after 90 days exposure to PBS at 37° C. In some non-fouling polymeric material comprises a homopolymer. embodiments, the modified surface exhibits a fibrinogen In an alternative preferred embodiment, the non-fouling poly adsorption of less than 20 ng/cm in a fibrinogen adsorption meric material comprises a random copolymer. In yet another assay in which samples are incubated for 60 minutes at 37°C. 45 embodiment, the non-fouling polymeric material comprises a in 70 g/mL fibrinogen derived from human plasma after 90 block copolymer, e.g., a diblock or triblock copolymer. days exposure to PBS at 37° C. In some embodiments, the In one embodiment the Surface modification, i.e., the modified surface exhibits a fibrinogen adsorption of less than grafted polymer, has a thickness which is at least equal to the 15 ng/cm in a fibrinogen adsorption assay in which samples surface roughness of the substrate surface. For example, if the are incubated for 60 minutes at 37°C. in 70 ug/mL fibrinogen 50 Surface of a substrate has a global average R. Surface rough derived from human plasma after 90 days exposure to PBS at ness of 100 nm, it is preferred in this embodiment that the 37°C. In some embodiments, the modified surface exhibits a grafted polymer layer have a global average dry thickness of fibrinogen adsorption of less than 12 ng/cm in a fibrinogen at least 100 nm. In some embodiments, the substrate surface adsorption assay in which samples are incubated for 60 min is relatively smooth, e.g., a global average R. Surface rough utes at 37°C. in 70 ug/mL fibrinogen derived from human 55 ness of 2 nm. In other embodiments, the Substrate Surface is plasma after 90 days exposure to PBS at 37° C. In some significantly rougher, e.g., a global average R. Surface embodiments, the modified surface exhibits a fibrinogen roughness of 1 Lum. In other embodiments, the Substrate Sur adsorption of less than 10 ng/cm in a fibrinogen adsorption face will have a surface roughness intermediate of these val assay in which samples are incubated for 60 minutes at 37°C. ues, e.g., a global average R. Surface roughness of 75-250 in 70 g/mL fibrinogen derived from human plasma after 90 60 nm. In each of these embodiments, it is preferred that the days exposure to PBS at 37° C. In some embodiments, the thickness of the grafted polymer layer exceed the global modified surface exhibits a fibrinogen adsorption of less than average R. Surface roughness of the Substrate surface. Thus, 8 ng/cm in a fibrinogen adsorption assay in which samples for example, in one embodiment the global average dry thick are incubated for 60 minutes at 37°C. in 70 ug/mL fibrinogen ness of the grafted polymer layer is at least 110% of the global derived from human plasma after 90 days exposure to PBS at 65 average R. Surface roughness of the Substrate surface. By 37°C. In some embodiments, the modified surface exhibits a way of further example, the global average dry thickness may fibrinogen adsorption of less than 6 ng/cm in a fibrinogen be at least 200% of the global average R. Surface roughness US 9,096,703 B2 29 30 of the substrate surface. By way of yet further example, the average R. Surface roughness of the modified surface is no global average dry thickness may be at least 500% of the more than the global average R. Surface roughness of the global average R. Surface roughness of the Substrate Sur surface of the article without the grafted polymer layer. face. By way of yet further example, the global average dry In one embodiment, and particularly for articles having thickness may be at least 1,000% of the global average R. Substrate Surfaces with relatively large surface roughness val Surface roughness of the Substrate Surface. In a preferred ues, the grafted polymer layer may reduce the Surface rough embodiment, the global average dry thickness of the grafted ness; stated differently, the modified surface, i.e., the surface polymer layer is determined using a scanning electron micro of the article with the grafted polymer, has less Surface rough scope (SEM) under vacuum and global average R. Surface ness than the Surface of the Substrate. For example, in one roughness is determined using an atomic force microscope. In 10 Such embodiment the global average R. Surface roughness one embodiment, the grafted polymer layer in each of the of the modified surface is at least 50% less than the global foregoing embodiments and examples recited in this para average R. Surface roughness of the Surface of the article graph is a Zwitterionic polymer. In one embodiment, the without the grafted polymer layer. By way of further example, grafted polymer in each of the foregoing examples and in one such embodiment the global average R. Surface embodiments recited in this paragraph is a polymer contain 15 roughness of the modified surface is at least 25% less than the ing carboxyammonium or Sulfoammonium repeat units. In global average R. Surface roughness of the surface of the one embodiment, the grafted polymer in each of the foregoing article without the grafted polymer layer. By way of further examples and embodiments recited in this paragraph is a example, in one such embodiment the global average R. polymer containing Sulfobetaine or carboxybetaine repeat surface roughness of the modified surface is at least 10% less units. In one embodiment, the grafted polymer in each of the than the global average R. Surface roughness of the Surface foregoing examples and embodiments recited in this para of the article without the grafted polymer layer. By way of graph is a Zwitterionic polymer and the Zwitterionic polymer further example, in one Such embodiment global average R. is grafted from a polyurethane polymer. In one embodiment, surface roughness of the modified surface is at least 5% less the grafted polymer in each of the foregoing examples and than the global average R. Surface roughness of the Surface embodiments recited in this paragraph is a carboxyammo 25 of the article without the grafted polymer layer. nium or Sulfoammonium polymer and the carboxyammo Independent of the relative surface roughness, the modified nium or Sulfoammonium polymer is grafted from a polyure Surface preferably has a relatively low surface roughness thane polymer. In one embodiment, the grafted polymer in value. For example, the modified surface preferably has a each of the foregoing examples and embodiments recited in global average R. Surface roughness of less than 500 nm. this paragraph is a polymer containing Sulfobetaine or car 30 By way of further example, the modified surface may have a boxybetaine repeat units and the polymer containing Sulfo global average R. Surface roughness of less than 400 nm. betaine or carboxybetaine repeat units is grafted from a poly By way of further example, the modified surface may have a urethane polymer. global average R. Surface roughness of less than 300 nm. In one embodiment, the grafted polymer layer preferen By way of further example, the modified surface may have a tially at least partially fills in defects in the substrate surface. 35 global average R. Surface roughness of less than 200 nm. Without being bound by any particular theory, a depression or By way of further example, the modified surface may have a invagination in the Substrate is Surrounded by initiator con global average R. Surface roughness of less than 150 nm. taining Substrate and therefore the amount of initiator avail By way of further example, the modified surface may have a able to drive polymerization within the depression or invagi global average R. Surface roughness of less than 100 nm. nation may be more Substantial than on a flat region of the 40 By way of further example, the modified surface may have a substrate. This may accelerate the polymerization in and fill global average R. Surface roughness of less than 75 nm. By ing of these defects. In some embodiments, defects in the way of further example, the modified surface may have a form of depressions having a depth of at least 100 nm as global average R. Surface roughness of less than 50 nm. By measured in a direction that is normal to the Surrounding way of further example, the modified surface may have a Surface and a width as measured in a direction parallel to and 45 global average R. Surface roughness of less than 25 nm. By at the surrounding surface that is at least 100 nm may be way of further example, the modified surface may have a preferentially filled with a grafted polymer. global average R. Surface roughness of less than 10 nm. By In one embodiment, the grafted polymer layer does not way of further example, the modified surface preferably has a significantly increase the Surface roughness. For example, in global average R. Surface roughness of less than 5 nm. By one embodiment, the modified surface, i.e., the surface of the 50 way of further example, the modified surface preferably has a article with the grafted polymer, has a Surface roughness global average R. Surface roughness of less than 2 nm. By value that is less than 300% of the global average R, surface way of further example, the modified surface preferably has a roughness of the surface of the article without the grafted global average R. Surface roughness of less than 1 nm. In polymer layer. By way of further example, in one Such one embodiment, the grafted polymer layer in each of the embodiment, the global average R. Surface roughness of 55 foregoing embodiments and examples recited in this para the modified surface is no more than 250% of the global graph is a Zwitterionic polymer. In one embodiment, the average R. Surface roughness of the Surface of the article grafted polymer in each of the foregoing examples and without the grafted polymer layer. By way of further example, embodiments recited in this paragraph is a polymer contain in one such embodiment, the global average R. Surface ing carboxyammonium or Sulfoammonium repeat units. In roughness of the modified surface is no more than 200% of 60 one embodiment, the grafted polymer in each of the foregoing the global average R. Surface roughness of the surface of examples and embodiments recited in this paragraph is a the article without the grafted polymer layer. By way of polymer containing Sulfobetaine or carboxybetaine repeat further example, in one such embodiment, the global average units. In one embodiment, the grafted polymer in each of the R. Surface roughness of the modified Surface is no more foregoing examples and embodiments recited in this para than 150% of the global average R. Surface roughness of the 65 graph is a Zwitterionic polymer and the Zwitterionic polymer surface of the article without the grafted polymer layer. By is grafted from a polyurethane polymer. In one embodiment, way of further example, in one such embodiment, the global the grafted polymer in each of the foregoing examples and US 9,096,703 B2 31 32 embodiments recited in this paragraph is a carboxyammo embodiment, the grafted polymer in each of the foregoing nium or Sulfoammonium polymer and the carboxyammo examples and embodiments recited in this paragraph is a nium or Sulfoammonium polymer is grafted from a polyure Zwitterionic polymer and the Zwitterionic polymer is grafted thane polymer. In one embodiment, the grafted polymer in from a polyurethane polymer. In one embodiment, the grafted each of the foregoing examples and embodiments recited in 5 polymer in each of the foregoing examples and embodiments this paragraph is a polymer containing Sulfobetaine or car recited in this paragraph is a carboxyammonium or Sulfoam boxybetaine repeat units and the polymer containing Sulfo monium polymer and the carboxyammonium or Sulfoammo betaine or carboxybetaine repeat units is grafted from a poly nium polymer is grafted from a polyurethane polymer. In one urethane polymer. embodiment, the grafted polymer in each of the foregoing In one embodiment, the grafted polymer layer may reduce 10 examples and embodiments recited in this paragraph is a the number of visual protrusions having a size greater than 0.1 polymer containing Sulfobetaine or carboxybetaine repeat micrometers relative to a reference Substrate, that is, the same units and the polymer containing Sulfobetaine or carboxybe or an otherwise functionally equivalent Substrate lacking the taine repeat units is grafted from a polyurethane polymer. non-fouling polymer layer. For example, the number of Such In general, the Surface modification preferably has a rela visual protrusions may be reduced by at least 25%. By way of 15 tively uniform thickness. For example, in one embodiment it further example, the number of Such visual protrusions may is generally preferred that the standard deviation of the global be reduced by at least 50%. By way of further example, the average dry thickness of the non-fouling grafted polymer number of such visual protrusions may be reduced by at least layer not exceed 100% of the global average dry thickness of 75%. By way of further example, the number of such visual the non-fouling grafted polymer layer. By way of further protrusions may be reduced by at least 90%. In one embodi example, in one embodiment the standard deviation of the ment, the grafted polymer layer may reduce the number of global average dry thickness of the non-fouling grafted poly visual protrusions having a size greater than 0.5 micrometers mer layer will not exceed 50% of the global average dry relative to a reference Substrate, that is, the same or an other thickness of the non-fouling grafted polymer layer By way of wise functionally equivalent Substrate lacking the non-foul further example, in one embodiment the standard deviation of ing polymer layer. For example, the number of Such visual 25 the global average dry thickness of the non-fouling grafted protrusions may be reduced by at least 25%. By way of further polymer layer will not exceed 20% of the global average dry example, the number of Such visual protrusions may be thickness of the non-fouling grafted polymer layer. By way of reduced by at least 50%. By way of further example, the further example, in one embodiment the standard deviation of number of such visual protrusions may be reduced by at least the global average dry thickness of the non-fouling grafted 75%. By way of further example, the number of such visual 30 polymer layer will not exceed 10% of the global average dry protrusions may be reduced by at least 90. thickness of the non-fouling grafted polymer layer. The stan Depending upon the article to which the surface modifica dard deviation of the thickness is preferably determined by tion is being applied and its working environment, the grafted taking at least 5, and more preferably at least 6-10, randomly polymer layer may have any of a wide range of thicknesses. spaced measurements of the grafted polymer layer thickness. For some applications, for example, the non-fouling grafted 35 In one embodiment, the grafted polymer in each of the fore polymer layer will have a global average dry thickness of at going examples and embodiments recited in this paragraph is least about 50 nm. For some applications, substantially a polymer containing carboxyammonium or Sulfoammonium thicker grafted polymer layers may be desirable. For repeat units. In one embodiment, the grafted polymer in each example, the non-fouling grafted polymer layer may have a of the foregoing examples and embodiments recited in this global average dry thickness of 50 micrometers. Typically, 40 paragraph is a polymer containing Sulfobetaine or carboxy however, the non-fouling grafted polymer layer will have an betaine repeat units. In one embodiment, the grafted polymer average thickness that is less. For example, in some embodi in each of the foregoing examples and embodiments recited in ments the non-fouling grafted polymer layer will have a glo this paragraph is a Zwitterionic polymer and the Zwitterionic balaverage dry thickness of up to 10 micrometers. By way of polymer is grafted from a polyurethane polymer. In one further example, in Some embodiments the non-fouling 45 embodiment, the grafted polymer in each of the foregoing grafted polymer layer will have a global average dry thickness examples and embodiments recited in this paragraph is a of up to 1 micrometer. By way of further example, in some carboxyammonium or Sulfoammonium polymer and the car embodiments the non-fouling grafted polymer layer will have boxyammonium or Sulfoammonium polymer is grafted from a global average dry thickness of up to 500 nm. By way of a polyurethane polymer. In one embodiment, the grafted further example, in Some embodiments the non-fouling 50 polymer in each of the foregoing examples and embodiments grafted polymer layer will have a global average dry thickness recited in this paragraph is a polymer containing Sulfobetaine in the range of about 100 nm to about 1,000 nm. By way of or carboxybetaine repeat units and the polymer containing further example, in Some embodiments the non-fouling sulfobetaine or carboxybetaine repeat units is grafted from a grafted polymer layer will have a global average dry thickness polyurethane polymer. in the range of about 300 nm to about 600 nm. By way of 55 In general, the Surface modifications of the present inven further example, in Some embodiments the non-fouling tion are relatively hydrophilic. In general, the modified sur grafted polymer layer will have a global average dry thickness face exhibits a static contactangle of less than 40 degrees. For in the range of about 200 nm to about 400 nm. In a preferred example, modified Surfaces of articles comprising non-foul embodiment, the global average dry thickness of the grafted ing polymeric materials of the present invention grafted from polymer layer is determined using a scanning electron micro- 60 a relatively hydrophobic polymer Such as silicone, hydrocar scope (SEM) under vacuum. In one embodiment, the grafted bon rubbers, fluorosilicones, fluoropolymers and other poly polymer in each of the foregoing examples and embodiments mers having a native contact angle of at least 90 degrees may recited in this paragraph is a polymer containing carboxyam exhibit a static contact angle of less than 40 degrees. By way monium or Sulfoammonium repeat units. In one embodiment, of further example, modified Surfaces of articles comprising the grafted polymer in each of the foregoing examples and 65 non-fouling polymeric materials of the present invention embodiments recited in this paragraph is a polymer contain grafted from a relatively hydrophobic Substrate having a con ing Sulfobetaine or carboxybetaine repeat units. In one tact angle of at least 90 degrees may exhibit a static contact US 9,096,703 B2 33 34 angle of less than 30 degrees. By way of further example, embodiment modified surfaces of articles having non-fouling modified surfaces of articles comprising non-fouling poly polymeric materials of the present invention grafted from a meric materials of the present invention grafted from a rela Substrate having a contact angle of at least 25 degrees exhibit tively hydrophobic Substrate having a contact angle of at least a static contact angle of less than 18 degrees. By way of 90 degrees may exhibit a static contact angle of less than 25 5 further example, in one embodiment modified surfaces of degrees. By way of further example, modified surfaces of articles having non-fouling polymeric materials of the articles having non-fouling polymeric materials of the present invention grafted from a Substrate having a contact present invention grafted from a relatively hydrophobic sub angle of at least 25 degrees exhibit a static contact angle of strate having a contactangle of at least 90 degrees may exhibit less than 17 degrees. By way of further example, in one a static contact angle of less than 20 degrees. By way of 10 embodiment modified surfaces of articles having non-fouling further example, modified Surfaces of articles having non polymeric materials of the present invention grafted from a fouling polymeric materials of the present invention grafted Substrate having a contact angle of at least 25 degrees exhibit from a relatively hydrophobic Substrate having a contact a static contact angle of less than 16 degrees. By way of angle of at least 90 degrees may exhibit a static contact angle further example, in one embodiment modified surfaces of of less than 15 degrees. In one embodiment, the grafted poly 15 articles having non-fouling polymeric materials of the mer in each of the foregoing examples and embodiments present invention grafted from a Substrate having a contact recited in this paragraph is a polymer containing carboxyam angle of at least 25 degrees exhibit a static contact angle of monium or Sulfoammonium repeat units. In one embodiment, less than 15 degrees. By way of further example, in one the grafted polymer in each of the foregoing examples and embodiment modified surfaces of articles having non-fouling embodiments recited in this paragraph is a polymer contain polymeric materials of the present invention grafted from a ing Sulfobetaine or carboxybetaine repeat units. In one Substrate having a contact angle of at least 25 degrees exhibit embodiment, the grafted polymer in each of the foregoing a static contact angle of about 5 to about 15 degrees. In one examples and embodiments recited in this paragraph is a embodiment, the non-fouling polymeric material in each of Zwitterionic polymer and the Zwitterionic polymer is grafted the foregoing embodiments and examples recited in this para from a polyurethane polymer. In one embodiment, the grafted 25 graph is a Zwitterionic polymer. In one embodiment, the polymer in each of the foregoing examples and embodiments grafted polymer in each of the foregoing examples and recited in this paragraph is a carboxyammonium or Sulfoam embodiments recited in this paragraph is a polymer contain monium polymer and the carboxyammonium or Sulfoammo ing carboxyammonium or Sulfoammonium repeat units. In nium polymer is grafted from a polyurethane polymer. In one one embodiment, the grafted polymer in each of the foregoing embodiment, the grafted polymer in each of the foregoing 30 examples and embodiments recited in this paragraph is a examples and embodiments recited in this paragraph is a polymer containing Sulfobetaine or carboxybetaine repeat polymer containing sulfobetaine or carboxybetaine repeat units. In one embodiment, the grafted polymer in each of the units and the polymer containing Sulfobetaine or carboxybe foregoing examples and embodiments recited in this para taine repeat units is grafted from a polyurethane polymer. graph is a Zwitterionic polymer and the Zwitterionic polymer Articles having non-fouling polymeric materials of the 35 is grafted from a polyurethane polymer. In one embodiment, present invention grafted from a less hydrophobic substrate the grafted polymer in each of the foregoing examples and Such as polyurethane (including aliphatic polycarbonate embodiments recited in this paragraph is a carboxyammo based polyurethanes) having a contact angle less than 90 nium or Sulfoammonium polymer and the carboxyammo degrees but greater than 25 degrees may exhibit a static con nium or Sulfoammonium polymer is grafted from a polyure tact angle of less than 25 degrees. For example, in one 40 thane polymer. In one embodiment, the grafted polymer in embodiment modified surfaces of articles having non-fouling each of the foregoing examples and embodiments recited in polymeric materials of the present invention grafted from a this paragraph is a polymer containing Sulfobetaine or car Substrate having a contact angle of at least 25 degrees exhibit boxybetaine repeat units and the polymer containing Sulfo a static contact angle of less than 24 degrees. By way of betaine or carboxybetaine repeat units is grafted from a poly further example, in one embodiment modified surfaces of 45 urethane polymer. articles having non-fouling polymeric materials of the In addition to being relatively hydrophilic, the grafted present invention grafted from a Substrate having a contact polymer layers of the present invention may also have a angle of at least 25 degrees exhibit a static contact angle of limited Swelling capacity. For example, in one embodiment less than 23 degrees. By way of further example, in one the difference between the dry thickness of the grafted poly embodiment modified surfaces of articles having non-fouling 50 merlayer and the thickness of the grafted polymer layer under polymeric materials of the present invention grafted from a ambient conditions is not great. For example, the magnitude Substrate having a contact angle of at least 25 degrees exhibit of the difference between the global average dry thickness of a static contact angle of less than 22 degrees. By way of the grafted polymer layer as determined by standard Scanning further example, in one embodiment modified surfaces of electron microscopy (SEM) to the global average humidified articles having non-fouling polymeric materials of the 55 thickness of the grafted polymer layer as determined by envi present invention grafted from a Substrate having a contact ronmental scanning electron microscopy (ESEM) is less than angle of at least 25 degrees exhibit a static contact angle of 200% of the global average dry thickness. For some applica less than 21 degrees. By way of further example, in one tions, even less Swelling capacity may be desirable. For embodiment modified surfaces of articles having non-fouling example, the difference in thickness of the grafted polymer polymeric materials of the present invention grafted from a 60 layer under such conditions may be less than 100% of the Substrate having a contact angle of at least 25 degrees exhibit global average dry thickness. By way of further example, the a static contact angle of less than 20 degrees. By way of difference in thickness of the grafted polymer layer as deter further example, in one embodiment modified surfaces of mined by SEM and ESEM under such conditions may be less articles having non-fouling polymeric materials of the than 50% of the global average dry thickness. By way of present invention grafted from a Substrate having a contact 65 further example, the difference in thickness of the grafted angle of at least 25 degrees exhibit a static contact angle of polymer layer as determined by SEM and ESEM under such less than 19 degrees. By way of further example, in one conditions may be less than 25% of the global average dry US 9,096,703 B2 35 36 thickness. By way of further example, the difference in thick with a global average dry thickness that is at least 200% of the ness of the grafted polymer layer as determined by SEM and global average R. Surface roughness of the substrate, a ESEM under such conditions may be less than 10% of the standard deviation for the thickness of the non-fouling grafted global average dry thickness. By way of further example, the polymer layer that does not exceed 50% of the global average difference in thickness of the grafted polymer layer as deter dry thickness of the non-fouling grafted polymer layer, and a mined by SEM and ESEM under such conditions may be less magnitude of the difference between the global average dry than 5% of the global average dry thickness. By way of thickness of the grafted polymer layer as determined by stan further example, the difference in thickness of the grafted dard Scanning electron microscopy (SEM) and the global polymer layer as determined by SEM and ESEM under such average humidified thickness of the grafted polymer layer as conditions may be less than 1% of the global average dry 10 determined by environmental scanning electron microscopy thickness. By way of further example, no difference may be (ESEM) that is less than 100% of the global average dry observable by Such a comparison. In one embodiment, the thickness. By way of further example, the process may be grafted polymer layer in each of the foregoing embodiments controlled to provide an article having a grafted polymer layer and examples recited in this paragraph is a Zwitterionic poly with a global average dry thickness that is at least 200% of the mer. In one embodiment, the grafted polymer in each of the 15 global average R. Surface roughness of the Substrate, a foregoing examples and embodiments recited in this para standard deviation for the thickness of the non-fouling grafted graph is a polymer containing carboxyammonium or Sul polymer layer that does not exceed 50% of the global average foammonium repeat units. In one embodiment, the grafted dry thickness of the non-fouling grafted polymer layer, and a polymer in each of the foregoing examples and embodiments magnitude of the difference between the global average dry recited in this paragraph is a polymer containing Sulfobetaine thickness of the grafted polymer layer as determined by stan or carboxybetaine repeat units. In one embodiment, the dard Scanning electron microscopy (SEM) and the global grafted polymer in each of the foregoing examples and average humidified thickness of the grafted polymer layer as embodiments recited in this paragraph is a Zwitterionic poly determined by environmental scanning electron microscopy mer and the Zwitterionic polymer is grafted from a polyure (ESEM) that is less than 50% of the global average dry thane polymer. In one embodiment, the grafted polymer in 25 thickness. By way of further example, the process may be each of the foregoing examples and embodiments recited in controlled to provide an article having a grafted polymer layer this paragraph is a carboxyammonium or Sulfoammonium with a global average dry thickness that is at least 200% of the polymer and the carboxyammonium or Sulfoammonium global average R. Surface roughness of the substrate, a polymer is grafted from a polyurethane polymer. In one standard deviation for the thickness of the non-fouling grafted embodiment, the grafted polymer in each of the foregoing 30 polymer layer that does not exceed 50% of the global average examples and embodiments recited in this paragraph is a dry thickness of the non-fouling grafted polymer layer, and a polymer containing sulfobetaine or carboxybetaine repeat magnitude of the difference between the global average dry units and the polymer containing Sulfobetaine or carboxybe thickness of the grafted polymer layer as determined by stan taine repeat units is grafted from a polyurethane polymer. dard Scanning electron microscopy (SEM) and the global Advantageously, the process of the present invention may 35 average humidified thickness of the grafted polymer layer as be tuned to provide independent control of the thickness, the determined by environmental scanning electron microscopy thickness uniformity, the degree of hydrophilicity (contact (ESEM) that is less than 25% of the global average dry angle), and/or the Swelling capacity of the grafted polymer thickness. By way of further example, the process may be layer, as well as the Surface roughness of the Surface-modified controlled to provide an article having a grafted polymer layer article. Thus, for example, the process may be controlled to 40 with a global average dry thickness that is at least 200% of the provide an article having a grafted polymer layer with a global average R. Surface roughness of the Substrate, a global average dry thickness that is at least 110% of the global standard deviation for the thickness of the non-fouling grafted average R. Surface roughness of the Substrate, a standard polymer layer that does not exceed 20% of the global average deviation for the thickness of the non-fouling grafted polymer dry thickness of the non-fouling grafted polymer layer, and a layer that does not exceed 100% of the global average dry 45 magnitude of the difference between the global average dry thickness of the non-fouling grafted polymer layer, and a thickness of the grafted polymer layer as determined by stan magnitude of the difference between the global average dry dard Scanning electron microscopy (SEM) and the global thickness of the grafted polymer layer as determined by stan average humidified thickness of the grafted polymer layer as dard Scanning electron microscopy (SEM) and the global determined by environmental scanning electron microscopy average humidified thickness of the grafted polymer layer as 50 (ESEM) that is less than 25% of the global average dry determined by environmental scanning electron microscopy thickness. By way of further example, the process may be (ESEM) that is less than 200% of the global average dry controlled to provide an article having a grafted polymer layer thickness. By way of further example, the process may be with a global average dry thickness that is at least 200% of the controlled to provide an article having a grafted polymer layer global average R. Surface roughness of the substrate, a with a global average dry thickness that is at least 200% of the 55 standard deviation for the thickness of the non-fouling grafted global average R. Surface roughness of the Substrate, a polymer layer that does not exceed 10% of the global average standard deviation for the thickness of the non-fouling grafted dry thickness of the non-fouling grafted polymer layer, and a polymer layer that does not exceed 50% of the global average magnitude of the difference between the global average dry dry thickness of the non-fouling grafted polymer layer, and a thickness of the grafted polymer layer as determined by stan magnitude of the difference between the global average dry 60 dard Scanning electron microscopy (SEM) and the global thickness of the grafted polymer layer as determined by stan average humidified thickness of the grafted polymer layer as dard Scanning electron microscopy (SEM) and the global determined by environmental scanning electron microscopy average humidified thickness of the grafted polymer layer as (ESEM) that is less than 25% of the global average dry determined by environmental scanning electron microscopy thickness. By way of further example, the process may be (ESEM) that is less than 200% of the global average dry 65 controlled to provide an article exhibiting a static contact thickness. By way of further example, the process may be angle of less than 25 degrees and a grafted polymer layer with controlled to provide an article having a grafted polymer layer a global average dry thickness that is at least 110% of the US 9,096,703 B2 37 38 global average R. Surface roughness of the Substrate, a controlled to provide an article exhibiting a static contact standard deviation for the thickness of the non-fouling grafted angle of less than 25 degrees and a grafted polymer layer with polymer layer that does not exceed 100% of the global aver a global average dry thickness that is at least 200% of the age dry thickness of the non-fouling grafted polymer layer, global average R. Surface roughness of the Substrate, a and a magnitude of the difference between the global average standard deviation for the thickness of the non-fouling grafted dry thickness of the grafted polymer layer as determined by polymer layer that does not exceed 50% of the global average standard Scanning electron microscopy (SEM) and the global dry thickness of the non-fouling grafted polymer layer, and a average humidified thickness of the grafted polymer layer as magnitude of the difference between the global average dry determined by environmental scanning electron microscopy thickness of the grafted polymer layer as determined by stan (ESEM) that is less than 200% of the global average dry 10 dard Scanning electron microscopy (SEM) and the global thickness. By way of further example, the process may be average humidified thickness of the grafted polymer layer as controlled to provide an article exhibiting a static contact determined by environmental scanning electron microscopy angle of less than 25 degrees and a grafted polymer layer with (ESEM) that is less than 10% of the global average dry a global average dry thickness that is at least 200% of the thickness. By way of further example, the process may be global average R. Surface roughness of the Substrate, a 15 controlled to provide an article exhibiting a static contact standard deviation for the thickness of the non-fouling grafted angle of less than 25 degrees and a grafted polymer layer with polymer layer that does not exceed 50% of the global average a global average dry thickness that is at least 200% of the dry thickness of the non-fouling grafted polymer layer, and a global average R. Surface roughness of the Substrate, a magnitude of the difference between the global average dry standard deviation for the thickness of the non-fouling grafted thickness of the grafted polymer layer as determined by stan polymer layer that does not exceed 50% of the global average dard Scanning electron microscopy (SEM) and the global dry thickness of the non-fouling grafted polymer layer, and a average humidified thickness of the grafted polymer layer as magnitude of the difference between the global average dry determined by environmental scanning electron microscopy thickness of the grafted polymer layer as determined by stan (ESEM) that is less than 100% of the global average dry dard Scanning electron microscopy (SEM) and the global thickness. By way of further example, the process may be 25 average humidified thickness of the grafted polymer layer as controlled to provide an article exhibiting a static contact determined by environmental scanning electron microscopy angle of less than 25 degrees and a grafted polymer layer with (ESEM) that is less than 5% of the global average dry thick a global average dry thickness that is at least 200% of the ness. By way of further example, the process may be con global average R. Surface roughness of the Substrate, a trolled to provide an article exhibiting a static contact angle of standard deviation for the thickness of the non-fouling grafted 30 less than 25 degrees and a grafted polymer layer with a global polymer layer that does not exceed 50% of the global average average dry thickness that is at least 200% of the global dry thickness of the non-fouling grafted polymer layer, and a average R. Surface roughness of the substrate, a standard magnitude of the difference between the global average dry deviation for the thickness of the non-fouling grafted polymer thickness of the grafted polymer layer as determined by stan layer that does not exceed 50% of the global average dry dard Scanning electron microscopy (SEM) and the global 35 thickness of the non-fouling grafted polymer layer, and a average humidified thickness of the grafted polymer layer as magnitude of the difference between the global average dry determined by environmental scanning electron microscopy thickness of the grafted polymer layer as determined by stan (ESEM) that is less than 50% of the global average dry dard Scanning electron microscopy (SEM) and the global thickness. By way of further example, the process may be average humidified thickness of the grafted polymer layer as controlled to provide an article exhibiting a static contact 40 determined by environmental scanning electron microscopy angle of less than 25 degrees and a grafted polymer layer with (ESEM) that is less than 5% of the global average dry thick a global average dry thickness that is at least 200% of the ness. By way of further example, in each of the foregoing global average R. Surface roughness of the Substrate, a examples, the grafted polymer layer may have a global aver standard deviation for the thickness of the non-fouling grafted age dry thickness in the range of 100 nm to 1,000 nm. polymer layer that does not exceed 50% of the global average 45 In general, grafted polymeric material may be detected in a dry thickness of the non-fouling grafted polymer layer, and a near-surface Zone of the substrate using EDS mapping, XPS, magnitude of the difference between the global average dry or TOF-SIMS. The sample may be freeze fractured in liquid thickness of the grafted polymer layer as determined by stan nitrogen to expose the coating/substrate interface. Fractured dard Scanning electron microscopy (SEM) and the global surface may then be coated with a thin layer of Au/Pt and average humidified thickness of the grafted polymer layer as 50 observed under a scanning electron microscope with Energy determined by environmental scanning electron microscopy Dispersive X-ray Analyser (EDAX) for element analysis. (ESEM) that is less than 25% of the global average dry Suitable instruments include a FEI/Philips XL30 FEG thickness. By way of further example, the process may be ESEM. In order to assess if the polymeric material extends controlled to provide an article exhibiting a static contact into the near-surface Zone, at least 25, and preferably at least angle of less than 25 degrees and a grafted polymer layer with 55 50, representative locations spaced approximately evenly a global average dry thickness that is at least 200% of the across the portion of the article carrying the grafted polymer global average R. Surface roughness of the Substrate, a layer should be analyzed to identify a detectable enhance standard deviation for the thickness of the non-fouling grafted ment of polymeric material in the near-surface Zone. For polymer layer that does not exceed 50% of the global average example, if a grafted polymer layer is applied to the indwell dry thickness of the non-fouling grafted polymer layer, and a 60 ing portion of a catheter, the representative locations are magnitude of the difference between the global average dry approximately evenly spaced across the indwelling portion of thickness of the grafted polymer layer as determined by stan the catheter. It is preferred to measure the thickness at repre dard Scanning electron microscopy (SEM) and the global sentative points across the longest dimension of the portion of average humidified thickness of the grafted polymer layer as the article that is covered with the grafted polymer layer. determined by environmental scanning electron microscopy 65 As described in greater detail elsewhere herein, incorpora (ESEM) that is less than 10% of the global average dry tion of initiator into the substrate enables polymeric material thickness. By way of further example, the process may be to be grafted from surface and from within near-surface Zone US 9,096,703 B2 39 40 of the substrate. In general, however, it is preferred that poly F1108, Ti-6Al-4V ELI (ASTM F136), Nitinol (ASTM meric material not extend too far into the Substrate; thus, in F2063), nickel titanium alloys, and thermo-memory alloy one embodiment polymeric material is present in the near materials; stainless steel (ASTM F138 and F139), tantalum Surface Zone but not at greater depths, i.e., not in the bulk. The (ASTM F560), palladium, zirconium, niobium, molybde maximum depth to which near-surface Zone extends is, at 5 num, nickel-chrome, or certain cobalt alloys including Stel least in part, a function of the initiator and the technique used lite, cobalt-chromium (Vitallium, ASTM F75 and Wrought to incorporate initiator in the substrate. Typically, however, it cobalt-chromium (ASTM F90)), and cobalt-chromium is generally preferred that lower boundary of the near-surface nickel alloys such as ELGILOYR, PHYNOX(R) and HAS Zone not be greater than 20 micrometers from the substrate TELLOYOR). Surface as measured in a direction normal to the Surface. By 10 Suitable ceramic materials include, but are not limited to, way of example, the lower boundary may not be greater than oxides, carbides, or nitrides of the transition elements such as 15 micrometers from the Substrate surface as measured in a titanium oxides, hafnium oxides, iridium oxides, chromium direction normal to the surface. By way of further example, oxides, aluminum oxides, and Zirconium oxides. Silicon the lower boundary may not be greater than 10 micrometers based materials, such as silica, may also be used. from the Substrate surface as measured in a direction normal 15 Suitable polymeric materials include, but are not limited to the surface. Similarly, the minimum depth of near-surface to, polyamide, polyamine, polyanhydride, polyazine, poly Zone, i.e., the distance of the upper boundary from the Sub (carbonate), polyester, polyether, polyetheretherketone strate Surface is, at least in part, also a function of the initiator (PEEK), polyguanidine, polyimide, polyketal, poly(ketone), and the technique used to incorporate initiator in the Sub polyolefin, poly(orthoester), polyphosphazine, polysaccha strate. Typically, however, the upper boundary will be at least ride, polysiloxane, polysulfone, polyurea, polyurethane, 0.1 micrometers from the Substrate Surface as measured in a halogenated polymer, silicone, aldehyde crosslinked resin, direction normal to the Surface. By way of example, the upper epoxy resin, phenolic resin, latex, or a copolymer or blend boundary may be at least 0.2 micrometers from the substrate thereof. Exemplary polymers include polystyrene and Substi Surface as measured in a direction normal to the Surface. By tuted polystyrenes, polyalkylenes, such as polyethylene and way of further example, the upper boundary may be at least 25 polypropylene, poly(urethane)S. polyacrylates and poly 0.3 micrometers from the substrate surface as measured in a methacrylates, polyacrylamides and polymethacrylamides, direction normal to the Surface. polyesters, polysiloxanes, polyethers (including polyac Substrates etals), poly(orthoesters), poly(carbonates), poly(hydroxyal In general, the Substrate comprises any of a wide range of kanoate)s, polyfluorocarbons, PEEK, Teflon, silicones, materials selected, for example, from one or more metals, 30 epoxy resins, KEVLARR, NOMEX(R), DACRONR), ceramics, glasses, polymers, biological tissues, living or HYTREL(R), PEBAX(R), SURLYNR), nylon, polyalkenes, dead, woven and non-woven fibers, semi-metals such as sili phenolic resins, PTFE, natural and synthetic elastomers, con, and combinations thereof. In one embodiment, the Sub adhesives and sealants, polyolefins, polysulfones, polyacry strate is a composite of two or more materials. For example, lonitrile, biopolymers such as polysaccharides and natural the Substrate may comprise a polymeric coating, also some 35 latex copolymers thereof, and combinations thereof. In one times referred to herein as an “undercoating.” or a “precoat embodiment the Substrate is a medical grade polyurethane ing over a metallic, ceramic, glass, polymeric, woven or such as CARBOTHANE(R), aliphatic polycarbonate-based non-woven fiber or semi-metal core. Alternatively, the sub polyurethanes, available from Lubrizol Corporation, blended strate may comprise a polymeric material throughout, i.e., with appropriate extrusion agents and plasticizers, possibly from its surface and into its bulk. By way of further example, 40 one already approved by the FDA or other appropriate regu the Substrate may comprise a polymeric coating, overlying a latory agency for use in vivo. Preferred substrates include metallic, ceramic, glass, polymeric, woven or non-woven elastollan, pearlthane, desmopan, estane, pellethane, irogan, fiber or semi-metal core inner layer which, in turn, overlies a exelast EC, laripur, carbothane, CARBOTHANER), isoplast, metallic, ceramic, glass, polymeric, woven or non-woven tecoflex, tecophilic, tecoplast, tecothane, biomer (Ethicon), fiber or semi-metal core. 45 biospan, cardiothane 51 (avothane), cardiomat, chronoflex As described in greater detail elsewhere herein, in a pre AL, chronoflex AR, chronoflex C, corplex, corethane, ferred embodiment of the polymerization method of the mitrathane, rimplast, toyobo TM5, vialon, enka PUR, com present invention, at least one polymerization initiator is feel ulcus, Viasorb, bioclusive, blisterfilm, opsite, tegaderm, incorporated into the substrate. As such, it is preferred that the epigard, lyofoam, omiderm, microthane, and Surethane. near-surface Zone of the Substrate comprise a material. Such 50 The Substrate may optionally contain a radiopaque addi as a polymer, having a sufficient capacity for the initiator(s). tive. Such as barium sulfate, bismuth salts, gold foil, or tan Thus, for example, when the Substrate comprises a metal, talum to aid in radiographic imaging. ceramic, glass or other material having insufficient capacity The substrate may be in the form of, or form part of gels, for initiator(s), the substrate is provided with an undercoat or liquids, films, particles (nanoparticles, microparticles, or mil a precoat having sufficient capacity for the polymerization 55 limeter diameter beads), fibers (wound dressings, bandages, initiator(s). gauze, tape, pads, sponges, including Woven and non-Woven In one embodiment, the Substrate may be a composite of sponges and those designed specifically for dental or oph two or more materials, e.g., an underlying material such as a thalmic Surgeries), blood storage bags, Surgical, medical or metal, ceramic, glass, semi-metal, polymer or other material dental instruments, blood oxygenators, ventilators, pumps, with a polymeric or other material coating thereon (e.g., an 60 drug delivery devices, tubing, wiring, electrodes, contracep undercoating or a precoating as described elsewhere herein). tive devices, feminine hygiene products, endoscopes, grafts In Such instances, the near-Surface Zone may lie partially (including Small diameter <6 mm), stents (including coro within the underlying material and partially within the poly nary, ureteral, renal, biliary, colorectal, esophageal, pulmo meric or other material coating thereon. nary, urethral, vascular, peripheral, neurovascular), Stent Suitable metallic materials include, but are not limited to, 65 grafts (including abdominal, thoracic, neurovascular and metals and alloys based on titanium, Such as unalloyed tita peripheral vascular), pacemakers, implantable cardioverter nium (ASTM F67) and titanium alloys, such as ASTM defibrillators, cardiac resynchronization therapy devices, car US 9,096,703 B2 41 42 diovascular device leads, Ventricular assist devices and driv coated with a medical grade polyurethane or CARBOTH elines, heart valves, Vena cava filters, endovascular coils, ANE(R). In another embodiment, the substrate is a vascularly catheters (including central venous, peripheral central, mid inserted catheter formed from a medical grade polyurethane line, peripheral, tunneled, dialysis access, urinary, neurologi or CARBOTHANER) containing a radiopaque additive, such cal, peritoneal, intra-aortic balloon pump, angioplasty bal as barium Sulfate or bismuth salts to aid in radiographic loon, diagnostic, interventional, drug delivery, etc.), catheter imaging, or formed from a material coated with a medical connectors and valves (including needleless connectors), grade polyurethane or CARBOTHANER) containing a radio intravenous delivery lines and manifolds, shunts (including paque additive, such as barium Sulfate or bismuth salts to aid cardiac, cerebral, lumbar-peritoneal, pulmonary, portosys in radiographic imaging. temic, portacaval, etc.), wound drains (internal or external 10 In another embodiment, the Substrate is a vascularly including ventricular, Ventriculoperitoneal, and lumboperito inserted catheter formed from a medical grade polyurethane neal), dialysis membranes, protein separation membranes, such as Tecothane(R) or formed from a material coated with a infusion ports, cochlear implants, endotracheal tubes, tra medical grade polyurethane Such as Tecothane(R). In another cheostomy tubes, ventilator breathing tubes and circuits, embodiment, the substrate is a vascularly inserted catheter guide wires, fluid collection bags, drug delivery bags and 15 formed from a medical grade polyurethane Such as Teco tubing, implantable sensors (e.g., intravascular, transdermal, thane(R) containing a radiopaque additive. Such as barium intracranial, glucose sensors), diagnostic devices (e.g., Sulfate or bismuth Salts to aid in radiographic imaging, or microfluidic, microelectromechanical, and optical), oph formed from a material coated with a medical grade polyure thalmic devices including contact lenses, intraocular lenses thane Such as Tecothane(R) containing a radiopaque additive, and phacoemulsification devices, orthopedic devices (includ Such as barium sulfate orbismuth salts, to aid in radiographic ing hip implants, knee implants, shoulder implants, spinal imaging. In another embodiment, the Substrate is a vascularly implants (including cervical plates Systems, pedicle screw inserted catheter formed from a medical grade polyurethane systems, interbody fusion devices, artificial disks, and other such as Pellethane(R) or formed from a material coated with a motion preservation devices), Screws, plates, rivets, rods, medical grade polyurethane such as Pellethane(R). In another intramedullary nails, bone cements, artificial tendons, and 25 embodiment, the substrate is a vascularly inserted catheter other prosthetics or fracture repair devices), dental implants, formed from a medical grade polyurethane such as Pelle periodontal implants, breast implants, penile implants, max thane(R) containing a radiopaque additive. Such as barium illofacial implants, cosmetic implants, valves, appliances, Sulfate or bismuth salts, to aid in radiographic imaging, or scaffolding, Suturing material, needles, hernia repair meshes, formed from a material coated with a medical grade polyure tension-free vaginal tape and vaginal slings, prosthetic neu 30 thane Such as Pellethane(R) containing a radiopaque additive, rological devices, tissue regeneration or cell culture devices, Such as barium sulfate orbismuth salts, to aid in radiographic dialyzer, cranial implants, syringes, blood collection contain imaging. ers, Scrotal implants, calve implants, buttock implants, Medical device substrates are often composed of multiple extraocular implants, horn implants, Subdermal implants, different materials, each with its own surface properties. Even transdermal implants, magnetic implants, medical devices 35 devices composed primarily of a single polymer may be made containing microfluidics, blood based sensors used outside of up of material blends and can include plasticizers, radio the body, nanoparticles used as sensors, IV catheter sheath, or opacity agents, and other additives all of which will affect other medical devices used within or in contact with the body Substrate Surface properties. To insure uniform Surface com or any portion of any of these. position for maximizing coating adhesion and efficacy, a The substrate may be in the form of, or form part of gels, 40 precoat of a single polymer or polymer blend may be placed foams, liquids, films, coatings, particles (nanoparticles, over the Substrate. In a particular embodiment, the undercoat microparticles, or millimeter diameter beads), fibers (includ ing coat contains a single polymer. ing woven and non-woven sponges and fabrics), marine and A polymer precoat or undercoat can be deposited on the underwater coatings (including coatings for ships, Subma Substrate using a variety of techniques known in the art, such rines, marine and hydrokinetic devices, aquariums, underwa 45 as solvent casting, dip-coating, spray-coating, plasma poly ter infrastructures, sewage pipes, and aqueduct tubes), pack merization, roller coating, electrostatic coating, or brush aging materials (including packaging for foods, beverages, coating. For example, the polymer to be applied as a precoat cosmetics, and consumer products), desalination and water or undercoat is dissolved in a solvent in which the substrate is treatment systems (including condensers, spacers, pipelines, substantially insoluble and the substrate is dipped therein to and membranes), separation membranes (including mem 50 deposit a layer of about 100 nm to about 500 micrometers of branes for macrofiltration, microfiltration, ultrafiltration, the precoat or undercoat polymer. Optionally, the deposited nanofiltration, and reversed osmosis filtration), lab appli polymer is crosslinked as it is being applied or after it has ances and consumer products including containers (e.g., petri been applied to the Substrate. Use of a single polymer under dishes, cell culture dishes, flasks, beakers), valves, needles, coating layer, for example, can result in the formation of a tapes, sealants, pipes, and tubes, earrings, body rings, contact 55 coating Surface that has a uniform identity and concentration lenses, cookware, gears (external/internal, spur, helical, of functional groups. double helical, bevel, hypoid, crown, worm, non-circular, In one preferred embodiment, a substrate is precoated with etc.), turbomachinary (turbines and compressors), pumps (di a polymer that conceals Substrate defects. The precoat thick rect lift, displacement, Velocity, buoyancy, and gravity), pro ness can be less than or greater than the amount of global pellers, blades, knives, windshields, and glassware. 60 average R. Surface roughness of the Substrate. In one pre In one embodiment, the Substrate is a vascularly inserted ferred embodiment, the Substrate has a precoat having an catheter Such as a peripherally inserted central catheter average thickness that exceeds the global average R. Sur (PICC), central venous catheter (CVC), or hemodialysis cath face roughness of the uncoated Substrate. As described else eter, venous valves, punctual plugs, and intra-ocular devices where herein, the precoating may optionally contain an ini and implants. In another embodiment, the Substrate is a vas 65 tiator or at least one member of an initiator pair. cularly inserted catheter formed from a medical grade poly In one embodiment, the surface of the substrate is treated to urethane or CARBOTHANER) or formed from a material improve the adherence of the precoat. For example, the sub US 9,096,703 B2 43 44 strate may be subjected to an oxidation pretreatment to monomer by means of a ring-opening polymerization reac increase the adhesion properties to the polymeric precoat; tion. Thus, the polymer may be a chain-growth homopolymer polymeric precoats may contain reactive groups that react or copolymer. In a preferred embodiment, the polymer is a with substrates forming a covalent bond. By way of further chain growth addition homopolymer or a chain growth addi example, prior to receiving a precoat, the Substrate may be tion copolymer comprising the residue of two or more mono silanized using Small molecule or polymeric reagents to CS. increase the adhesion properties to the polymeric precoat. By In accordance with one aspect of the present invention, it is way of further example, the surface may be subjected to generally preferred that the non-fouling polymeric material alternating organic and aqueous treatments. be prepared without inordinate use of a polyfunctional The undercoating layer may contain a radiopaque agent, 10 crosslinking agent. For example, it is generally preferred that Such as BaSO or bismuth, to aid in radiographic imaging of the Substrate. In one embodiment the polymer is a polyure the non-fouling polymeric material contain less than 50 mole thane polymer such as Tecoflex-93A or Carbothane 85A, % of the residue of a polyvalent crosslinker. In one such optionally containing 0 to 40% by weight BaSO. embodiment, the non-fouling polymeric material contains The undercoating layer can also include, but is not limited 15 less than 25 mole% of the residue of a polyvalent crosslinker. to, polymers such as polystyrene and Substituted polysty In one such embodiment, non-fouling polymeric material renes, polyethylene, polypropylene, poly(urethane)S. poly containless than 10 mole% of a polyvalent crosslinker. In one acrylates and polymethacrylates, polyacrylamides and poly Such embodiment, the non-fouling polymeric material con methacrylamides, polyesters, polysiloxanes, polyethers, poly tains less than 5 mole % of the residue of a polyvalent (orthoester), poly(carbonates), poly(hydroxyalkanoate)S. crosslinker. In one Such embodiment, non-fouling polymeric polyfluorocarbons, PEEK, Teflon, silicones, epoxy resins, material contain less than 3 mole % of a polyvalent KEVLARR, NOMEX(R), DACRONR), HYTRELR), crosslinker. In one Such embodiment, the non-fouling poly PEBAX(R), SURLYNR), nylon, polyalkenes, phenolic resins, meric material contains less than 0.1 mole'/6 of the residue of PTFE, natural and synthetic elastomers, adhesives and seal a polyvalent crosslinker. In one such embodiment, the non ants, polyolefins, polysulfones, polyacrylonitrile, biopoly 25 fouling polymeric material contains no residue of a polyva merS Such as polysaccharides and natural latex copolymers lent crosslinker. thereof, and combinations thereof. Through grafting, step-growth or chain-growth tech The precoated substrate can then be further functionalized niques, the non-fouling polymeric material may comprise any using the coating methods described below. of a range of polymer types or combinations thereof. The In the case where a greater density of non-fouling material 30 polymer backbone may be neutral (e.g., polyalkylene or poly is desired, the creation of microstructure on the substrate ether) or contain permanently charged moieties (e.g., cyclic surface can create more area for grafting non-fouling mate or acyclic quaternized nitrogen atoms), or even Zwitterionic rials from the Surface, without increasing the apparent Surface backbones (e.g., phosphorylcholine backbones). In one area of the article. For polymeric Substrates, including hydro embodiment, therefore, the non-fouling polymeric material gel networks, this Surface morphology can be created through 35 comprises a polymer or copolymer selected from the group appropriate polymer structural design. One example of this consisting of polyamide, polyamine, polyanhydride, methodology is the growth of surface tethered dendrimeric polyazine, poly(carbonate), polyester, polyether, polyethere polymers. Each generation of the dendrimer effectively therketone (PEEK), polyguanidine, polyimide, polyketal, doubles the number of Zwitterionic sites presenting. Other poly(ketone), polyolefin, poly(orthoester), polyphosphazine, polymer architectures include brush polymers, such as brush 40 polysaccharide, polysiloxane, polysulfone, polyurea, poly copolymers, comb polymers, such as comb copolymers, lin urethane, halogenated polymer, silicone, hydrocarbon, ether ear and branched copolymers, crosslinked polymers, hydro ester, ether-amide or ionized polyethylene and combinations gels, polymer blends, and combinations thereof. thereof. Surface Modifications The polymer may also contain a wide range of pendant In general, a non-fouling polymeric material is grafted 45 (side-chain) groups, hydrophilic and hydrophobic, neutral, from a substrate into which one or more polymerization ini anionic, cationic, or mixed charged. For example, the pendant tiators have been incorporated. In one embodiment, the non groups may include neutral hydrophilic groups such as fouling polymeric material is grafted from a Substrate that is hydroxy, oligo(ethylene glycol) and/or poly(ethylene glycol) a composite of two or more materials, e.g., an underlying moieties, or it may include charged groups such as anionic material Such as a metal, ceramic, glass, semi-metal, polymer 50 moieties, cationic moieties, and Zwitterionic moieties. or other material with a polymeric or other material coating Zwitterionic Groups thereon (e.g., an undercoating or a precoating as previously Zwitterions are molecules that carry formal positive and described herein). For example, in one embodiment, the non negative charges on non-adjacentatoms within the same mol fouling polymeric material is grafted from a polymeric under ecule and molecules that may be ionized by addition or coat layer, Such as a polyurethane layer which overlies a metal 55 removal of an electrophile or a nucleophile, or by removal of or ceramic bulk. By way of further example, in one embodi a protecting group. Both natural and synthetic polymers, con ment the non-fouling polymeric material is grafted from a taining Zwitterion functionality, have been shown to resist polymeric undercoat layer, Such as a polyurethane layer protein adhesion. In one embodiment, the Zwitterionic mono which overlies a polymeric bulk, such as polyurethane. mer contains a phosphorylcholine moiety, a carboxyammo Preferably, the non-fouling polymeric material that is 60 nium moiety, a Sulfoammonium moiety, derivatives thereof, grafted from the Substrate comprises a chain-growth polymer or combinations thereof. In one embodiment, the Zwitterionic (that is, a polymer or polymer block formed by addition monomer contains a carboxyammonium moiety, a Sulfoam polymerization), or a combination thereof. The chain-growth monium moiety, derivatives thereof, or combinations thereof. polymer may be, for example, an addition polymer derived In one embodiment, the Zwitterionic monomer contains a from monomer(s) incorporating double or triple bonds, e.g., 65 sulfobetaine moiety or a carboxybetaine moiety. The Zwitte an olefin. By way of further example, the chain-growth poly rionic polymer may be formed by initiating polymerization mer may comprise an addition polymer derived from a cyclic with radicals present in the polymeric Substrate, in the pres US 9,096,703 B2 45 46 ence of one or more monomers, such as Sulfobetaine meth bination with T and the nitrogen atom to which they are acrylate or carboxybetaine methacrylate monomers. attached form a nitrogen-containing heteroaromatic ring, Polysulfoammonium polymers such as polysulfobetaines, T'' is hydrocarbylene, substituted hydrocarbylene, ether, or oxylated alkylene, polycarboxyammonium polymers such as polycarboxybe Z is carboxylate, phosphate, phosphonic, phosphonate, taines and other natural and synthetic Zwitterion chemistries Sulfate, Sulfinic, or Sulfonate, and can be used to design non-fouling materials for the biomedi * designates the point of covalent attachment, direct or cal applications described herein. Some examples of natural indirect, of the Zwitterion of Formula ZI-3 to the polymer Zwitterions chemistries that could be used for non-fouling backbone. materials include, but are not limited to, amino acids, pep In certain preferred embodiments in which the polymer tides, natural Small molecules including, but not limited to, 10 contains Zwitterionic pendant group corresponding to For N.N.N-trimethylglycine (glycine betaine), trimethylamine mula ZI-3, T, T, T', and T' are selected from a more oxide (TMAO), dimethylsulfoniopropionate sarcosine, lyser narrow range of substituents, Z is carboxylate or sulfate, and gic acid and psilocybin. Additional synthetic Zwitterions that the Zwitterion corresponds to Formula ZI-4: could be used to create non-fouling materials, include, but are not limited to, amino-carboxylic acids (carboxybetaines), 15 amino-Sulfonic acids (sulfo betaines), cocamidopropyl Formula ZI-4 betaine, quinonoid based Zwitterions, decaphenylferrocene, T13 and non-natural amino acids. Natural and synthetic polymers T (T ed also include mixed charged structures with both positive 1 s NZ charged and negative charged moieties on the pendant T14 groups, in the main chains, or at the terminal groups. Materials containing, or composed of these natural or wherein * designates the point of covalent attachment, direct synthetic Zwitterions, can be grafted from Surfaces, particu or indirect, of the Zwitterion of Formula ZI-4 to the polymer larly the surfaces of medical devices, in order to improve 25 backbone; T is a bond or -(CH), with m being 1 to 3: biocompatibility, reduce thrombogenesis (Such as on the Sur T' and T''' are independently hydrogen, alkyl, or substituted face of stents or venous valves), and reduce fouling by pro alkyl: T is optionally substituted alkylene, phenylene, ether, teins or bacteria present in solution. This is particularly appli or oxylated alkylene; and Z is carboxylate or sulfate. For cable for Surfaces where non-specific binding of proteins in example, in this embodiment, T'and T''may independently Solution could negatively impact the desired or necessary 30 be hydrogen or lower alkyl, e.g., methyl, ethyl, or propyl. By mechanics of a device. way of further example, in this embodiment, T'and T''may In one embodiment, the non-fouling polymer contains independently be hydrogen or lower alkyl, e.g., methyl, ethyl, Zwitterionic pendant groups covalently attached, directly or or propyl. By way of further example, in this embodiment, indirectly to the polymer backbone. The Zwitterionic pendant T' may be —(CH), with n being 1-8. By way of further groups may have an overall net charge, for instance, by having 35 example, in this embodiment, T' may be -(CH2) - or a divalent center of anionic charge and monovalent center of —(CH2) and T and T'' may be methyl. By way of cationic charge or vice versa, or by having two centers of further example, in this embodiment, T''maybe —(CH) - cationic charge and one centerofanionic charge or vice versa. or —(CH) , T and T'' may be hydrogen or alkyl. By Preferably, however, the Zwitterion has no overall net charge way of further example, in this embodiment, T' may be and most preferably has a center of monovalent cationic 40 —(CH) , T' and T' may be methyl, T' may be charge and a center of monovalent anionic charge. Addition —(CH2) and Z may be carboxylate. By way of further ally, the center(s) of cationic charge are preferably perma example, in this embodiment, T' may be —(CH) , T' nent; that is, it is preferably a quaternary nitrogen, quaternary and T''may be methyl, T' may be —(CH), and Z' may phosphonium or tertiary Sulfonium group. Additionally, the be sulfate. center(s) of anionic charge are also permanent; that is, they 45 In certain preferred embodiments in which the polymer are completely ionized at physiological pH and are preferably contains Zwitterionic pendant group corresponding to For carboxylate, phosphate, phosphonic, phosphonate, Sulfate, mula ZI-3, T, Tand T'and the nitrogenatom to which they Sulfinic, or Sulfonate. are attached form a nitrogen-containing heteroaromatic ring. In another embodiment, the polymer contains Zwitterionic For example, T, Tand T' and the nitrogen atom to which pendant groups covalently attached, directly or indirectly, to 50 they are attached may form an optionally substituted hetero the polymer back bone, and the Zwitterion corresponds to cycle, containing a quaternary nitrogen atom. One Such Formula ZI-3: embodiment corresponds to Formula ZI-5:

Formula ZI-3 55 Formula ZI-5 T9 G) T5 e 1 T8Se- TI N. G -HET1Nz4 lo wherein * designates the point of covalent attachment, direct 60 or indirect, of the Zwitterion of Formula ZI-5 to the polymer wherein backbone; HET is a heterocycle containing a quaternary T is a bond, hydrocarbylene, substituted hydrocarbylene, nitrogen atom, T is optionally substituted alkylene, phe heterocyclo, or in combination with Tand T'and the nitro nylene, ether, or oxylated alkylene; and Z is carboxylate or gen atom to which they are attached form a nitrogen-contain sulfate. For example, in this embodiment, T' may be ing heteroaromatic ring, 65 —(CH), with n being 1-8. By way of further example, in Tand T' are independently hydrogen, hydrocarbyl, sub this embodiment, T''maybe -(CH2) - or-(CH2)4- and stituted hydrocarbyl or heterocyclo, or, Tand T', in com Z may be carboxylate or sulfate. By way of further example, US 9,096,703 B2 47 48 in this embodiment, T' may be —(CH2) and Z' may be * designates the point of covalent attachment, direct or sulfate. By way of further example, in this embodiment, T' indirect, of the Zwitterion of Formula ZI-1 to the polymer may be -(CH2) and Z may be carboxylate. Exemplary backbone. Zwitterions corresponding to Formula ZI-5 include Zwitteri In certain preferred embodiments in which the polymer ons corresponding to Formulae ZI-6A and ZI-6B: contains Zwitterionic pendant group corresponding to For mula ZI-1, T and T are oxygen, Z' is quaternary nitrogen, and the Zwitterion corresponds to Formula ZI-2: Formula ZI-6A

10 Formula ZI-2 G T4 5 Formula ZI-6B O O 1.T 1n 1 n T3 n T6

15 O wherein * designates the point of covalent attachment, direct wherein * designates the point of covalent attachment of the or indirect, of the Zwitterion of Formulae ZI-6A and ZI-6B to Zwitterion of Formula ZI-2 to the polymer backbone, T is the polymer backbone; T' is optionally substituted alkylene, hydrocarbylene, substituted hydrocarbylene, or oxylated phenylene, ether, or oxylated alkylene; and Z' is carboxylate alkylene, and T, T and T are independently hydrogen, or sulfate. For example, in this embodiment, T' may be hydrocarbyl, substituted hydrocarbyl or heterocyclo. For —(CH), with n being 1-8. By way of further example, in example, in this embodiment, T may be -(CH2), with n this embodiment, T' may be —(CH), or—(CH), and being 1-8. By way of further example, in this embodiment, Z may be carboxylate or sulfate. By way of further example, T, Tand T may independently be lower alkyl, e.g., methyl, in this embodiment, T' may be —(CH2) and Z' may be 25 ethyl or propyl. By way of further example, in this embodi sulfate. By way of further example, in this embodiment, T' ment, T may be —(CH), with n being 1-3, and T.T and may be -(CH2)2 - and Z' may be carboxylate. T may independently be lower alkyl, e.g., methyl, ethyl or In one embodiment, the polymer contains Zwitterionic propyl. By way of further example, in this embodiment, T pendant groups covalently attached, directly or indirectly, to may be -(CH2), with n being 1-3, and one or more of T. the polymer back bone, and the Zwitterion corresponds to 30 Tand T may be substituted hydrocarbyl such as oligomeric Formula ZI-7 phosphorylcholine (e.g., Formula 9). Neutral Hydrophilic Pendant Groups In one embodiment, the polymer contains neutral hydro Formula ZI-7 philic pendant groups covalently attached, directly or indi 35 rectly, to the polymer backbone. Exemplary neutral hydro philic groups include hydroxy, thiol, oxylated alkyls (e.g., oligoethylene glycol, polyethylene glycol and/or polypropy lene glycol), ether, thioether, and the like. In one Such specific embodiment, the polymer contains pendant groups compris 40 ing alkoxylated moieties corresponding to Formula POA-1: wherein T, Tand T are independently hydrogen, hydrocar Formula POA-1 byl, substituted hydrocarbyl or heterocyclo: T' is a bond, R2 hydrocarbylene, substituted hydrocarbylene, or heterocyclo, 45 : O and * designates the point of covalent attachment, direct or R3 indirect, of the Zwitterion of Formula ZI-7 to the polymer backbone. R1 In one embodiment, the polymer contains Zwitterionic pendant groups covalently attached, directly or indirectly, to 50 wherein a is 1-3, b is 1-8, each R' and R is independently the polymer back bone, and the Zwitterion corresponds to selected from the group consisting of hydrogen, halogen, and Formula ZI-1: optionally substituted lower alkyl, R is hydrocarbyl, substi tuted hydrocarbyl or heterocyclo, and * designates the point of attachment of the moieties corresponding to Formula Formula ZI-1 55 O POA-1 to the remainder of the pendant group and the back O GE) bone. By way of example, in one such embodiment, each R' TN-Tš Zl and Rare hydrogen, n is 2 or 3. By way of further example, 1n 1 N-1 in one such embodiment, each RandR is hydrogen, n is 2 or | O 3, and b is 3-5. By way of further example, in one such 60 embodiment, each RandR is hydrogen, n is 2 or 3, b is 3-5, and R is alkyl. In one embodiment, the repeat units are wherein derived from macromonomers containing 2-20 alkylene T and T are independently oxygen, sulfur, NH or a bond, oxide units. T is hydrocarbylene, substituted hydrocarbylene, ether, or Repeat Units oxylated alkylene, 65 In general, homopolymers or copolymers comprising Zwit Z' is a moiety comprising a quaternary nitrogen, phospho terionic pendant groups, neutral hydrophilic pendant groups, nium or Sulfonium cationic group, and cationic pendant groups and/or anionic pendant groups may US 9,096,703 B2 49 50 be prepared by polymerization of any of a wide range of monomers. In one preferred embodiment, the non-fouling Formula 3 polymeric material is a homopolymer or copolymer compris ing repeat units derived from an olefinic monomer. Thus, for example, in one embodiment the non-fouling polymeric 5 material comprises repeat units derived from an olefinic monomer and corresponding to Formula 1: wherein X comprises an oxylated alkylene moiety, a zwit Formula 1 10 terionic moiety, an anionic moiety, or a cationic moiety. For XI X3 example, X may be OX, NXX or SX, C-C wherein X* is a substituted hydrocarbylor heterocyclo moi ety comprising an oxylated alkylene moiety, a Zwitterionic X2 X4 moiety, an anionic moiety, or a cationic moiety, and X* is 15 hydrogen, hydrocarbyl, substituted hydrocarbyl or heterocy wherein clo. For example, X may behydrogen or lower alkyl. By way X' and X’ are independently hydrogen, hydrocarbyl, sub of further example, X may be OX, or - NHX". By stituted hydrocarbyl, heterocyclo, or substituted carbonyl, way of further example, X may be OX', or -NHX" provided, however, X" and X’ are not each selected from the wherein X comprises an oxylated alkylene moiety corre group consisting of aryl, heteroaryl, and heterosubstituted sponding to Formula POA-1. By way of further example, X." carbonyl, may be —OX', or -NHX' wherein X comprises a zwit X is hydrogen, alkyl or substituted alkyl, terionic moiety corresponding to Formula ZI-1, ZI-2, ZI-3, X4 is OX10, NX41X42, N*X41X42x43, SX40, ZI-4, ZI-5, ZI-6A, ZI-6B, or ZI-7. By way of further example, aryl, heteroaryl or acyl, 25 the repeat unit of Formula 3 may be a cationic repeat unit. By X" is hydrogen, hydrocarbyl, substituted hydrocarbyl, way of further example, the repeat unit of Formula 3 may be heterocyclo or acyl, and an anionic repeat unit. By way of further example, X may be X', X and X’ are independently hydrogen, hydrocar hydrogen or methyl and X may comprise an oxylated alky byl, substituted hydrocarbyl or heterocyclo. lene moiety corresponding to Formula POA-1 or a Zwitteri 30 onic moiety corresponding to Formula ZI-1, ZI-2, ZI-3, ZI-4, In certain embodiments in which the non-fouling poly ZI-5, ZI-6A, ZI-6B, or ZI-7. In one particularly preferred meric material comprises repeat units corresponding to For embodiment, the polymer contains repeat units correspond mula 1, it is preferred that X of at least a fraction of the repeat ing to Formula 3 and X* is —O(CH)N(CH)(CH), units comprise alkoxylated moieties, Zwitterionic moieties, SO, O(CH)N(CH)(CH2)CO, NH(CH)N" anionic moieties, or cationic moieties. In Such embodiments, 35 (CH)(CH) CO, or - NH(CH)N(CH)(CH),SO, for example, X' and X may be hydrogen, and the polymer wherein n is 1-8. In one embodiment, the polymer contains comprises repeat units corresponding to Formula 2: repeat units corresponding to Formula 3 and X* is NH (CH), N(CH2)CH(CH2)SO, NH(CH), N(CH2), CH(CH2)CO. NH(CH2)N(CH2)CH)2(CH2)SOs, Formula 2 40 NH(CH.)N(CH2)CH,), (CH) CO, NH(CH),N- cyclo-(CH2)CO, or NH(CH2)..Ncyclo-(CH2)SOs, (Ncyclo is a heterocyclic structure or a heterocyclic derivative containing at least one nitrogen element), wherein m is 1-8; in is 0-5; and p is 1-8. In one embodiment, the polymer contains 45 repeat units corresponding to Formula 3 and X is wherein X is hydrogen, alkyl or substituted alkyl, and X* is O(CH2)N(CH2)CH-(CH2)SOs, O(CH2)N(CH2), a pendant group comprising an oxylated alkylene moiety, a CH,(CH2)CO, O(CH),N" (CH), CH,-(CH2)SO, Zwitterionic moiety, an anionic moiety, or a cationic moiety. O(CH2)N(CH2)CHI (CH2)CO, O(CH2)..Ncy For example, X may be hydrogen or lower alkyl. By way of clo-(CH2)CO, or O(CH.)..Ncyclo-(CH.)SO, wherein 50 m is 1-8; n is 0-5; and p is 1-8. In one embodiment, the further example, X may be a pendant group comprising an polymer contains repeat units corresponding to Formula 3 oxylated alkylene moiety corresponding to Formula POA-1. and X* is O(CH),N-(CH)(CH)SO = O(CH)N" By way of further example, the repeat unit of Formula 2 may (CH)(CH2)CO. —NH(CH)N(CH)(CH2)SO, be Zwitterionic repeat unit comprising a Zwitterionic moiety NH(CH)N(CH)(CH) CO, NH(CH)N(CH), corresponding to Formula ZI-1, ZI-2, ZI-3, ZI-4, ZI-5, ZI-6A, 55 (CH)SO, NH(CH)N(CH)(CH2)CO. —O(CH) ZI-6B, or ZI-7. By way of further example, the repeat unit of N"(CHCH)(CH)SO. —O(CH)N(CHCH)(CH), Formula 2 may be a cationic repeat unit. By way of further CO. —O(CH)N(CH2CH2CHCH) (CH)SO, example, the repeat unit of Formula 2 may be an anionic - O(CH)N(CHCHCHCH)(CH),CO, or - NH repeat unit. By way of further example, X may be hydrogen (CH)Ncyclo-(CH2)SO. or methyl and X may be a pendant group comprising an 60 In one preferred embodiment, the non-fouling polymeric oxylated alkylene moiety corresponding to Formula POA-1 material is a Zwitterionic polymer or copolymer. For example, ora Zwitterionic moiety corresponding to Formula ZI-1, ZI-2, the non-fouling polymeric material may comprise carboxy ZI-3, ZI-4, ZI-5, ZI-6A, ZI-6B, or ZI-7. betaine repeat units and/or Sulfobetaine repeat units. Alterna In one presently preferred embodiment, the non-fouling tively, the non-fouling polymeric material may be a polyam polymeric material comprises repeat units corresponding to 65 pholyte, containing anionic and cationic repeat units. Formula 2 wherein X is acyl and the repeat units correspond Optionally, the non-fouling polymer may contain poly(ethyl to Formula 3: ene oxide) repeat units and/or other neutral olefinic repeat US 9,096,703 B2 51 52 units. Thus, for example, in one preferred embodiment, the one embodiment the sum of a and c is at least 0.2 and d is at non-fouling polymeric material is a Zwitterionic polymer or least 0.1. By way of further example, in one embodiment the copolymer comprising the repeat units of Formula 4: sum of a and c is at least 0.3 and d is at least 0.1. By way of further example, in one embodiment the Sum of a and c is at least 0.4 and d is at least 0.1. By way of further example, in Formula 4 one embodiment the sum of a and c is at least 0.5 and d is at A B C D least 0.1. By way of further example, in one embodiment the X3 X3 X3 X3 sum of a and c is at least 0.6 and d is at least 0.1. By way of H H H H further example, in one embodiment the Sum of a and c is at -1 --C C --C d 10 least 0.7 and d is at least 0.1. By way of further example, in X4 one embodiment the sum of a and c is at least 0.8 and d is at (O or NH) O (O or NH) 'O (O or NH) 'O least 0.1. By way of further example, in one embodiment the sum of a and c is at least 0.9 and d is at least 0.1. By way of pi further example, in one embodiment the Sum of a and c is at 15 least 0.1, b is at least 0.1 and d is at least 0.1. By way of further O -N- example, in one embodiment the Sum of a and c is at least 0.2, 2s. b is at least 0.1 and d is at least 0.1. By way of further example, in one embodiment the sum of a and c is at least 0.3, b is at p least 0.1 and d is at least 0.1. By way of further example, in OY O O 29 G one embodiment the Sum of a and c is at least 0.4, b is at least 2,1 0.1 and d is at least 0.1. By way of further example, in one O embodiment the sum of a and c is at least 0.5, b is at least 0.1 and d is at least 0.1. By way of further example, in one a is 0-1 b is 0-1; c is 0-1; d is 0-1; m is 1-20; n and o are embodiment the sum of a and c is at least 0.6, b is at least 0.1, independently 0-11; p and q are independently 0-11: X is 25 and d is at least 0.1. By way of further example, in one hydrogen, alkyl or substituted alkyl, X is —OX'. embodiment the sum of a and c is at least 0.7, b is at least 0.1 - NX'X', SX', aryl, heteroaryl or acyl: X" is hydro and d is at least 0.1. By way of further example, in one gen, hydrocarbyl, substituted hydrocarbyl, heterocyclo or embodiment the sum of a and c is at least 0.8, b is at least 0.1 acyl: X' and X’ are independently hydrogen, hydrocarbyl, and d is at least 0.1. By way of further example, in one substituted hydrocarbyl or heterocyclo; and X* is hydrogen, 30 embodiment the sum of a and c is at least 0.9, b is at least 0.1 hydrocarbyl or substituted hydrocarbyl, provided the sum of and d is at least 0.1. In each of these exemplary embodiments, a, b, c and d is greater than 0 and X of repeat unit D differs a may be 0, c may be 0, or a and c may each be greater than 0. from the corresponding pendant group of repeat units A, B In one preferred embodiment, the non-fouling polymeric and C. In one such embodiment, X is hydroxy-substituted material is a Zwitterionic polymer or copolymer comprising alkyl Such as hydroxypropyl. 35 the repeat units of Formula 4 m is 1-8; X is hydrogen, alkyl In one embodiment, it is preferred that the non-fouling or substituted alkyl, X* is OX', NX'X', SX', aryl, polymeric material is a Zwitterionic polymer comprising heteroaryl or acyl: X" is hydrogen, hydrocarbyl, substituted repeat units corresponding to the A and/or the C repeat units. hydrocarbyl, heterocyclo or acyl: X' and X’ are indepen For example, in one embodiment the Sum of a and c is at least dently hydrogen, hydrocarbyl, substituted hydrocarbyl or 0.1. By way of further example, in one embodiment the sum 40 heterocyclo; and X* is hydrogen, hydrocarbyl or substituted ofa and c is at least 0.2. By way of further example, in one hydrocarbyl, with the proviso that X of the D repeat differs embodiment the sum of a and c is at least 0.3. By way of from the corresponding pendant groups of the A, B or C further example, in one embodiment the Sum of a and c is at repeat units and a, b, c, and d, in combination, are selected least 0.4. By way of further example, in one embodiment the from one of the sets of combinations appearing in Table I: sum of a and c is at least 0.5. By way of further example, in 45 one embodiment the sum of a and c is at least 0.6. By way of further example, in one embodiment the Sum of a and c is at least 0.7. By way of further example, in one embodiment the sum of a and c is at least 0.8. By way of further example, in one embodiment the sum of a and c is at least 0.9. By way of 50 further example, in one embodiment the Sum of a and c is at least 0.1 and b is at least 0.1. By way of further example, in one embodiment the Sum of a and c is at least 0.2 and b is at least 0.1. By way of further example, in one embodiment the sum of a and c is at least 0.3 and b is at least 0.1. By way of 55 further example, in one embodiment the Sum of a and c is at least 0.4 and b is at least 0.1. By way of further example, in one embodiment the sum of a and c is at least 0.5 and b is at least 0.1. By way of further example, in one embodiment the sum of a and c is at least 0.6 and b is at least 0.1. By way of 60 further example, in one embodiment the Sum of a and c is at least 0.7 and b is at least 0.1. By way of further example, in one embodiment the sum of a and c is at least 0.8 and b is at least 0.1. By way of further example, in one embodiment the sum of a and c is at least 0.9 and b is at least 0.1. By way of 65 further example, in one embodiment the Sum of a and c is at least 0.1 and d is at least 0.1. By way of further example, in US 9,096,703 B2 54 repeat units corresponding of Formula 4 are cationic repeat units (X for such units is a cationic pendant group). By way of further example, in one such embodiment, d is at least 0.2 and approximately one-half the repeat units corresponding to repeat unit D are anionic repeat units (X for such units is an anionic pendant group) and approximately one-half of the repeat units corresponding of Formula 4 are cationic repeat units (X for such units is a cationic pendant group). By way of further example, in one such embodiment, d is at least 0.3 10 and approximately one-half the repeat units corresponding to repeat unit D are anionic repeat units (X for such units is an anionic pendant group) and approximately one-half of the repeat units corresponding of Formula 4 are cationic repeat 15 units (X for such units is a cationic pendant group). By way of further example, in one such embodiment, d is at least 0.4 and approximately one-half the repeat units corresponding to repeat unit D are anionic repeat units (X for such units is an anionic pendant group) and approximately one-half of the repeat units corresponding of Formula 4 are cationic repeat units (X for such units is a cationic pendant group). By way of further example, in one such embodiment, d is at least 0.5 and approximately one-half the repeat units corresponding to repeat unit D are anionic repeat units (X for such units is an 25 anionic pendant group) and approximately one-half of the repeat units corresponding of Formula 4 are cationic repeat units (X for such units is a cationic pendant group). By way of further example, in one such embodiment, d is at least 0.6 and approximately one-half the repeat units corresponding to 30 repeat unit D are anionic repeat units (X for such units is an anionic pendant group) and approximately one-half of the repeat units corresponding of Formula 4 are cationic repeat units (X for such units is a cationic pendant group). By way of further example, in one such embodiment, d is at least 0.7 35 and approximately one-half the repeat units corresponding to repeat unit D are anionic repeat units (X for such units is an anionic pendant group) and approximately one-half of the repeat units corresponding of Formula 4 are cationic repeat 40 units (X for such units is a cationic pendant group). By way of further example, in one such embodiment, d is at least 0.8 and approximately one-half the repeat units corresponding to repeat unit D are anionic repeat units (X for such units is an anionic pendant group) and approximately one-half of the 45 repeat units corresponding of Formula 4 are cationic repeat units (X for such units is a cationic pendant group). By way of further example, in one such embodiment, d is at least 0.9 and approximately one-half the repeat units corresponding to repeat unit D are anionic repeat units (X for such units is an 50 anionic pendant group) and approximately one-half of the repeat units corresponding of Formula 4 are cationic repeat units (X for such units is a cationic pendant group). By way of further example, in each of said examples in this paragraph, the remaining repeat units may correspond to repeat unit A. 55 By way of further example, in each of said examples in this In one embodiment, the non-fouling polymeric material is paragraph, the remaining repeat units may correspond to a polyampholyte Zwitterionic polymer or copolymer com repeat unit B. By way of further example, in each of said prising repeat units corresponding to repeat unit D of Formula examples in this paragraph, the remaining repeat units may 4. That is, d is greater than 0 and a fraction of the repeat units correspond to repeat unit C. corresponding to repeat unit Dare anionic repeat units (X" for 60 More preferably, the non-fouling polymeric material is a Such units is an anionic pendant group) and a fraction of the Zwitterionic polymer or copolymer comprising repeat units repeat units corresponding of Formula 4 are cationic repeat corresponding to repeat unit A and/or repeat unit C of For units (X for such units is a cationic pendant group). For mula 4. example, in one such embodiment, d is at least 0.1 and In certain embodiments, the non-fouling polymeric mate approximately one-half the repeat units corresponding to 65 rial is a homopolymer or copolymer comprising repeat units repeat unit D are anionic repeat units (X for such units is an corresponding to Formula 5. Formula 6, Formula 7. Formula anionic pendant group) and approximately one-half of the 8, or Formula 9: US 9,096,703 B2 56

Formula 5 Formula 10 X3 --X x X4 Formula 6 X7 lo Formula 11 X6 - 10 X8 Formula 7 --X-HET-- Formula 8 X10 X13 15 Formula 12 X9 X12 XII X14 Formula 9 O X7 X8 | le. -to--o- -- -- wherein X is hydrocarbylene, substituted hydrocarbylene, O X8 heterocyclo, amide, anhydride, ester, imide, thioester, thioet G her, urethane, or urea; X7 is hydrogen, alkyl or substituted 25 alkyl; and X is an anionic moiety. Suitable comonomers include, but are not limited to, acry lates, acrylamides, vinyl compounds, multifunctional mol ecules, such as di-, tri-, and tetraisocyanates, di-, tri-, and HET is part of a heterocyclic structure, 30 tetraols, di-, tri-, and tetraamines, and di-, tri-, and tetrathio X is hydrogen, alkyl or substituted alkyl, cyanates; cyclic monomers, such as lactones and lactams, and X is OX, NX'X', SX', aryl, heteroaryl or combination thereof. In the interests of brevity, exemplary acyl, methacrylate monomers are listed below (but it should be understood that analogous acrylate, acrylamide and meth X is ester, anhydride, imide, amide, ether, thioether, 35 acrylamide monomers may be similarly listed and are simi thioester, hydrocarbylene, substituted hydrocarbylene, het larly included): erocyclo, urethane, or urea; Charged methacrylates or methacrylates with primary, sec X is hydrocarbylene, substituted hydrocarbylene, hetero ondary or tertiary amine groups, such as, 3-sulfopropyl cyclo, amide, anhydride, ester, imide, thioester, thioether, methacrylate potassium salt, (2-dimethylamino)ethyl 40 methacrylate) methyl chloride quaternary salt, 2 urethane, or urea; (methacryloyloxy)ethyltrimethyl-ammonium chloride, X is hydrogen, alkyl or substituted alkyl: methacryloyl chloride, 3-(methacryloylamino)propyl X is an anionic moiety; trimethylammonium chloride), 2-aminoethyl methacry X is hydrocarbylene, substituted hydrocarbylene, hetero late hydrochloride, 2-(diethylamino)ethyl methacrylate, 45 2-(dimethylamino)ethyl methacrylate, 2-(tert-buty cyclo, amide, anhydride, ester, imide, thioester, thioether, lamino)ethyl methacrylate, and 2-(tert-butylamino urethane, or urea; ethyl methacrylate. X" is hydrogen, alkyl or substituted alkyl: Alkyl methacrylates or other hydrophobic methacrylates, X' is a cationic moiety; Such as ethyl methacrylate, butyl methacrylate, hexyl 50 methacrylate, 2-ethylhexyl methacrylate, methyl meth X' is hydrocarbylene, substituted hydrocarbylene, hetero acrylate, lauryl methacrylate, isobutyl methacrylate, cyclo, amide, anhydride, ester, imide, thioester, thioether, isodecyl methacrylate, phenyl methacrylate, decyl urethane, or urea; methacrylate, 3,3,5-trimethylcyclohexyl methacrylate, X' is hydrogen, alkyl or substituted alkyl: benzyl methacrylate, cyclohexyl methacrylate, Stearyl 55 methacrylate, tert-butyl methacrylate, tridecyl meth X'' is an anionic moiety; acrylate, 2-naphthyl methacrylate, 2.2.3,3-tetrafluoro L' and L are independently hydrocarbylene, substituted propyl methacrylate, 1.1.1.3.3.3-hexafluoroisopropyl hydrocarbylene, heterocyclo, amide, anhydride, ester, imide, methacrylate, 2.2.2-trifluoroethyl methacrylate, 2.2.3,3, thioester, thioether, urethane, or urea; and 3-pentafluoropropyl methacrylate, 2.2.3.4.4.4- X" is hydrogen, hydrocarbyl, substituted hydrocarbyl, 60 hexafluorobutyl methacrylate, 2.2.3.3.4.4.4-heptafluo heterocyclo or acyl, and robutyl methacrylate, 2.2.3.3.4.4.5.5-octafluoropentyl methacrylate, 3.3.4.4.5,5,6,6,7,7,8,8,8-tridecafluorooc X' and X’ are independently hydrogen, hydrocarbyl, tyl methacrylate, and 3.3.4.4.5,5,6,6,7,7,8,8,9,9,10,10. substituted hydrocarbyl or heterocyclo. 10-heptadecafluorodecyl methacrylate. In one embodiment, the non-fouling polymeric material 65 Reactive or crosslinkable methacrylates, such as 2-(trim comprises repeat units corresponding to Formula 7 wherein ethylsilyloxy)ethyl methacrylate, 3-(trichlorosilyl)pro the heterocycle. HET corresponds to Formulae 10, 11 or 12: pyl methacrylate, 3-(trimethoxysilyl)propyl methacry US 9,096,703 B2 57 58 late, 3-tris(trimethylsiloxy)silylpropyl methacrylate, grafted polymer layer. In some preferred embodiments, the trimethylsilyl methacrylate, allyl methacrylate, vinyl grafted polymer layer contains XPS signals of nitrogen, and methacrylate, 3-(acryloyloxy)-2-hydroxypropyl meth optionally sulfur. acrylate, 3-(diethoxymethylsilyl)propyl methacrylate In general, the grafted polymeric material may comprise 3-(dimethylchlorosilyl)propyl methacrylate 2-isocy 5 repeat units corresponding to any of Formulae 1 to 12. By way anatoethyl methacrylate, glycidyl methacrylate, 2-hy of further example, the grafted polymeric material may com droxyethyl methacrylate, 3-chloro-2-hydroxypropyl prise a Zwitterionic polymer. By way of further example, methacrylate, Hydroxybutyl methacrylate, glycol meth polymeric material may comprise repeat units corresponding acrylate, hydroxypropyl methacrylate, and 2-hydrox to Formula 1. By way of further example, the grafted poly ypropyl 2-(methacryloyloxy)ethyl phthalate. 10 meric material may comprise repeat units corresponding to Formula 2. By way of further example, the grafted polymeric Other methacrylates, such as ethylene glycol methyl ether material may comprise repeat units corresponding to Formula methacrylate, di(ethylene glycol) methyl ether meth 3. By way of further example, the grafted polymeric material acrylate, ethylene glycol phenyl ether methacrylate, may comprise repeat units corresponding to Formula 4. Addi 2-butoxyethyl methacrylate, 2-ethoxyethyl methacry 15 tionally, the grafted polymeric material may comprise, as late, and ethylene glycol dicyclopentenyl ether meth pendant groups, any of the pendant groups disclosed herein. acrylate. Thus, for example, the grafted polymeric material may com Multifunctional monomers, such as di, tri, or tetraacrylates prise pendant groups corresponding to any of Formulae ZI-1 and di, tri, or tetraacrylamides can be used to form highly to ZI-7 or POA-1. In one particularly preferred embodiment, branched structures which can provide a higher concentration the grafted polymeric material corresponds to Formula 1 and of non-fouling groups on the Surface. As previously noted, the comprises Zwitterionic pendant groups. In another particu non-fouling polymeric material may contain a non-Zwitteri larly preferred embodiment, the grafted polymeric material onic non-fouling material, alone or in combination with a corresponds to Formula 3 and comprises Sulfobetaine or car Zwitterionic material. These non-fouling groups may have boxybetaine pendant groups. In one especially preferred varying degrees of non-fouling performance in a range of 25 embodiment, the grafted polymeric material comprises environments. Suitable non-Zwitterionic materials include, repeat units derived from sulfobetaine methacrylate, sulfobe but are not limited to, polyethers, such as polyethylene glycol, taine acrylate, Sulfobetaine acrylamide, Sulfobetaine meth poly(ethylene oxide-co-propylene oxide) (PEO-PPO) block acrylamide, carboxybetaine methacrylate, carboxybetaine copolymers, polysaccharides such as dextran, hydrophilic acrylate, carboxybetaine acrylamide, or carboxybetaine polymers such as polyvinylpyrrolidone (PVP) and hydroxy 30 methacrylamide monomers. In general, the height and any ethyl-methacrylate (HEMA), acrylonitrile-acrylamide branching of the grafted polymeric material can help to over copolymers, heparin, heparin fragments, derivatized heparin come surface irregularities and defects, and increased branch fragments, hyaluronic acid, mixed charge materials, and ing may reduce the ability of fouling materials to penetrate the materials containing hydrogen bond accepting groups. Such non-fouling layer. as those described in U.S. Pat. No. 7,276.286. Suitable poly 35 In one preferred embodiment, the grafted polymeric mate mer structures included, but are not limited to, polymers or rial corresponds to Formula 1 and comprises Zwitterionic copolymers containing monomers of Formula I wherein ZI is pendant groups and the Surface modification has a thickness replaced by a non-Zwitterionic, non-fouling head group. which is at least equal to the Surface roughness of the Sub In one embodiment, the non-fouling material is a polymer strate Surface. In one Such preferred embodiment, the grafted containing repeat units derived from Sulfobetaine-containing 40 polymeric material corresponds to Formula 3 and comprises and/or carboxybetaine-containing monomers. Examples of Sulfobetaine or carboxybetaine pendant groups. In one Such monomers include sulfobetaine methacrylate (SBMA), sul preferred embodiment, the grafted polymeric material com fobetaine acrylamide, Sulfobetaine methacrylamide, car prises repeat units derived from sulfobetaine methacrylate, boxybetaine methacrylate (CBMA), carboxybetaine acryla sulfobetaine acrylate, sulfobetaine acrylamide, sulfobetaine mide and carboxybetaine methacrylamide. Examples of Such 45 methacrylamide, carboxybetaine methacrylate, carboxybe polymers include, but are not limited to, poly(carboxy betaine taine acrylate, carboxybetaine acrylamide, or carboxybetaine methacrylate) (polyCBMA), poly(carboxybetaine acryla methacrylamide monomers and has a global average dry mide), poly(carboxybetaine methacrylamide) poly(Sulfobe thickness that is at least 110% of the global average R. taine methacrylate) (polySBMA), poly(sulfobetaine acryla Surface roughness of the Substrate Surface. In one such pre mide), and poly(Sulfobetaine methacrylamide). In another 50 ferred embodiment, the grafted polymeric material is a embodiment, the non-fouling material polymer is a polymer homopolymer of sulfobetaine methacrylate, sulfobetaine containing the residue of CBMA or SBMA and one or more acrylate, Sulfobetaine acrylamide, Sulfobetaine methacryla additional monomers. The additional monomers can be Zwit mide, carboxybetaine methacrylate, carboxybetaine acrylate, terionic or non-Zwitterionic monomers. carboxybetaine acrylamide, or carboxybetaine methacryla In some embodiments, it is preferred to have use Zwitteri 55 mide and has a global average dry thickness that is at least onic polymers that possess permanently charged groups, 200% of the global average R. Surface roughness of the which, without being bound by any theory, may improve substrate surface. In one such preferred embodiment, the non-fouling performance because the charged groups are grafted polymeric material is a copolymer, at least 50% of the ionically solvated with water. The presence of commonly monomeric residues of which are residues of sulfobetaine used groups which can have permanent charges in the Zwit 60 methacrylate, Sulfobetaine acrylate, Sulfobetaine acrylamide, terionic polymers can be detected by using XPS to analyze the Sulfobetaine methacrylamide, carboxybetaine methacrylate, elements present in the top approximately 1-50 nm of the carboxybetaine acrylate, carboxybetaine acrylamide, or car Surface. One representative group commonly used in Zwitte boxybetaine methacrylamide and has a global average dry rions is nitrogen in quaternary amine groups. In Sulfobetaine, thickness that is at least 200% of the global average R. elemental signal of nitrogen may be approximately equiva 65 Surface roughness of the Substrate Surface. In one such pre lent to a signal for sulfur. Further, techniques such as TOF ferred embodiment, the grafted polymeric material is a SIMS may be used to identify Zwitterionic groups in the copolymer, at least 60% of the monomeric residues of which US 9,096,703 B2 59 60 are residues of sulfobetaine methacrylate, sulfobetaine acry vacuum. In one such preferred embodiment, the grafted poly late, Sulfobetaine acrylamide, Sulfobetaine methacrylamide, meric material is a copolymer, at least 70% of the monomeric carboxybetaine methacrylate, carboxybetaine acrylate, car residues of which are residues of sulfobetaine methacrylate, boxybetaine acrylamide, or carboxybetaine methacrylamide sulfobetaine acrylate, sulfobetaine acrylamide, sulfobetaine and has a global average dry thickness that is at least 200% of 5 methacrylamide, carboxybetaine methacrylate, carboxybe the global average R. Surface roughness of the Substrate taine acrylate, carboxybetaine acrylamide, or carboxybetaine Surface. In one such preferred embodiment, the grafted poly methacrylamide and has a global average dry thickness of at meric material is a copolymer, at least 70% of the monomeric least about 50 nm, as measured by SEMunder vacuum. In one residues of which are residues of sulfobetaine methacrylate, Such preferred embodiment, the grafted polymeric material is sulfobetaine acrylate, sulfobetaine acrylamide, sulfobetaine 10 a copolymer, at least 80% of the monomeric residues of which methacrylamide, carboxybetaine methacrylate, carboxybe are residues of sulfobetaine methacrylate, sulfobetaine acry taine acrylate, carboxybetaine acrylamide, or carboxybetaine late, Sulfobetaine acrylamide, Sulfobetaine methacrylamide, methacrylamide and has a global average dry thickness that is carboxybetaine methacrylate, carboxybetaine acrylate, car at least 200% of the global average R. Surface roughness of boxybetaine acrylamide, or carboxybetaine methacrylamide the substrate surface. In one such preferred embodiment, the 15 and has a global average dry thickness of at least about 50 nm, grafted polymeric material is a copolymer, at least 80% of the as measured by SEM under vacuum. In one such preferred monomeric residues of which are residues of sulfobetaine embodiment, the grafted polymeric material is a copolymer, methacrylate, Sulfobetaine acrylate, Sulfobetaine acrylamide, at least 90% of the monomeric residues of which are residues Sulfobetaine methacrylamide, carboxybetaine methacrylate, of sulfobetaine methacrylate, sulfobetaine acrylate, sulfobe carboxybetaine acrylate, carboxybetaine acrylamide, or car taine acrylamide, Sulfobetaine methacrylamide, carboxybe boxybetaine methacrylamide and has a global average dry taine methacrylate, carboxybetaine acrylate, carboxybetaine thickness that is at least 200% of the global average R. acrylamide, or carboxybetaine methacrylamide and has a glo Surface roughness of the Substrate Surface. In one Such pre balaverage dry thickness of at least about 50 nm, as measured ferred embodiment, the grafted polymeric material is a by SEM under vacuum. By way of further example, in each of copolymer, at least 90% of the monomeric residues of which 25 the foregoing embodiments, the global average dry thickness are residues of sulfobetaine methacrylate, sulfobetaine acry may be even greater, e.g., at least about 200 nm, at least about late, Sulfobetaine acrylamide, Sulfobetaine methacrylamide, 300 nm, at least about 400 nm, or at least about 500 nm. carboxybetaine methacrylate, carboxybetaine acrylate, car In another preferred embodiment, the grafted polymeric boxybetaine acrylamide, or carboxybetaine methacrylamide material corresponds to Formula 1 and comprises Zwitteri and has a global average dry thickness that is at least 200% of 30 onic pendant groups and the Surface modification, i.e., the the global average R. Surface roughness of the Substrate grafted polymeric material, has a relatively uniform thick surface. ness. In one such preferred embodiment, the grafted poly In another preferred embodiment, the grafted polymeric meric material corresponds to Formula 3 and comprises Sul material corresponds to Formula 1 and comprises Zwitteri fobetaine or carboxybetaine pendant groups. In one Such onic pendant groups and the Surface modification, i.e., the 35 preferred embodiment, the grafted polymeric material com grafted polymeric material, has a global average dry thick prises repeat units derived from sulfobetaine methacrylate, ness of at least 50 nm. In one such preferred embodiment, the sulfobetaine acrylate, sulfobetaine acrylamide, sulfobetaine grafted polymeric material corresponds to Formula 3 and methacrylamide, carboxybetaine methacrylate, carboxybe comprises Sulfobetaine or carboxybetaine pendant groups. In taine acrylate, carboxybetaine acrylamide, or carboxybetaine one Such preferred embodiment, the grafted polymeric mate 40 methacrylamide monomers. In one such preferred embodi rial comprises repeat units derived from sulfobetaine meth ment, polymeric material is a homopolymer of Sulfobetaine acrylate, Sulfobetaine acrylate, Sulfobetaine acrylamide, Sul methacrylate, Sulfobetaine acrylate, Sulfobetaine acrylamide, fobetaine methacrylamide, carboxybetaine methacrylate, Sulfobetaine methacrylamide, carboxybetaine methacrylate, carboxybetaine acrylate, carboxybetaine acrylamide, or car carboxybetaine acrylate, carboxybetaine acrylamide, or car boxybetaine methacrylamide monomers. In one such pre 45 boxybetaine methacrylamide monomers and the standard ferred embodiment, polymeric material is a homopolymer of deviation of the global average dry thickness of the non sulfobetaine methacrylate, sulfobetaine acrylate, sulfobe fouling grafted polymer layer not exceed 100% of the global taine acrylamide, Sulfobetaine methacrylamide, carboxybe average dry thickness of the non-fouling grafted polymer taine methacrylate, carboxybetaine acrylate, carboxybetaine layer. In one such preferred embodiment, the grafted poly acrylamide, or carboxybetaine methacrylamide monomers 50 meric material is a copolymer, at least 50% of the monomeric and has a global average dry thickness of at least about 50 nm, residues of which are residues of sulfobetaine methacrylate, as measured by SEM under vacuum. In one such preferred sulfobetaine acrylate, sulfobetaine acrylamide, sulfobetaine embodiment, the grafted polymeric material is a copolymer, methacrylamide, carboxybetaine methacrylate, carboxybe at least 50% of the monomeric residues of which are residues taine acrylate, carboxybetaine acrylamide, or carboxybetaine of sulfobetaine methacrylate, sulfobetaine acrylate, sulfobe 55 methacrylamide and the standard deviation of the global aver taine acrylamide, Sulfobetaine methacrylamide, carboxybe age dry thickness of the non-fouling grafted polymer layer taine methacrylate, carboxybetaine acrylate, carboxybetaine not exceed 100% of the global average dry thickness of the acrylamide, or carboxybetaine methacrylamide and has a glo non-fouling grafted polymer layer. In one Such preferred balaverage dry thickness of at least about 50 nm, as measured embodiment, the grafted polymeric material is a copolymer, by SEM under vacuum. In one such preferred embodiment, 60 at least 60% of the monomeric residues of which are residues the grafted polymeric material is a copolymer, at least 60% of of sulfobetaine methacrylate, sulfobetaine acrylate, sulfobe the monomeric residues of which are residues of sulfobetaine taine acrylamide, Sulfobetaine methacrylamide, carboxybe methacrylate, Sulfobetaine acrylate, Sulfobetaine acrylamide, taine methacrylate, carboxybetaine acrylate, carboxybetaine Sulfobetaine methacrylamide, carboxybetaine methacrylate, acrylamide, or carboxybetaine methacrylamide and the stan carboxybetaine acrylate, carboxybetaine acrylamide, or car 65 dard deviation of the global average dry thickness of the boxybetaine methacrylamide and has a global average dry non-fouling grafted polymer layer not exceed 100% of the thickness of at least about 50 nm, as measured by SEM under global average dry thickness of the non-fouling grafted poly US 9,096,703 B2 61 62 mer layer. In one such preferred embodiment, the grafted late, Sulfobetaine acrylamide, Sulfobetaine methacrylamide, polymeric material is a copolymer, at least 70% of the mono carboxybetaine methacrylate, carboxybetaine acrylate, car meric residues of which are residues of sulfobetaine meth boxybetaine acrylamide, or carboxybetaine methacrylamide acrylate, Sulfobetaine acrylate, Sulfobetaine acrylamide, Sul and the modified surface exhibits a static contact angle of less fobetaine methacrylamide, carboxybetaine methacrylate, than 25 degrees. In one such preferred embodiment, the carboxybetaine acrylate, carboxybetaine acrylamide, or car grafted polymeric material is a copolymer, at least 70% of the boxybetaine methacrylamide and the standard deviation of monomeric residues of which are residues of sulfobetaine the global average dry thickness of the non-fouling grafted methacrylate, Sulfobetaine acrylate, Sulfobetaine acrylamide, polymer layer not exceed 100% of the global average dry Sulfobetaine methacrylamide, carboxybetaine methacrylate, thickness of the non-fouling grafted polymer layer. In one 10 carboxybetaine acrylate, carboxybetaine acrylamide, or car Such preferred embodiment, the grafted polymeric material is boxybetaine methacrylamide and the modified surface exhib a copolymer, at least 80% of the monomeric residues of which its a static contact angle of less than 25 degrees. In one Such are residues of sulfobetaine methacrylate, sulfobetaine acry preferred embodiment, the grafted polymeric material is a late, Sulfobetaine acrylamide, Sulfobetaine methacrylamide, copolymer, at least 80% of the monomeric residues of which carboxybetaine methacrylate, carboxybetaine acrylate, car 15 are residues of sulfobetaine methacrylate, sulfobetaine acry boxybetaine acrylamide, or carboxybetaine methacrylamide late, Sulfobetaine acrylamide, Sulfobetaine methacrylamide, and the standard deviation of the global average dry thickness carboxybetaine methacrylate, carboxybetaine acrylate, car of the non-fouling grafted polymer layer not exceed 100% of boxybetaine acrylamide, or carboxybetaine methacrylamide the global average dry thickness of the non-fouling grafted and the modified surface exhibits a static contact angle of less polymer layer. In one such preferred embodiment, the grafted than 25 degrees. In one such preferred embodiment, the polymeric material is a copolymer, at least 90% of the mono grafted polymeric material is a copolymer, at least 90% of the meric residues of which are residues of sulfobetaine meth monomeric residues of which are residues of sulfobetaine acrylate, Sulfobetaine acrylate, Sulfobetaine acrylamide, Sul methacrylate, Sulfobetaine acrylate, Sulfobetaine acrylamide, fobetaine methacrylamide, carboxybetaine methacrylate, Sulfobetaine methacrylamide, carboxybetaine methacrylate, carboxybetaine acrylate, carboxybetaine acrylamide, or car 25 carboxybetaine acrylate, carboxybetaine acrylamide, or car boxybetaine methacrylamide and the standard deviation of boxybetaine methacrylamide and the modified surface exhib the thickness of the non-fouling grafted polymer layer not its a static contact angle of less than 25 degrees. By way of exceed 100% of the global average dry thickness of the non further example, in each of the foregoing embodiments, the fouling grafted polymer layer. By way of further example, in modified Surface exhibits a static contact angle may be even each of the foregoing embodiments, the standard deviation of 30 less, e.g., less than 24, less than 23, less than 22, less than 21, thickness may be even less, e.g., less than 50% of the global less than 20, less than 19, less than 18, less than 17, less than average dry thickness of the non-fouling grafted polymer 16, or less than 15. layer, less than 20% of the global average dry thickness of the In another preferred embodiment, the grafted polymeric non-fouling grafted polymer layer, or less than 10% of the material corresponds to Formula 1, comprises Zwitterionic global average dry thickness of the non-fouling grafted poly 35 pendant groups and the grafted polymeric material, i.e., the mer layer. grafted polymer layer, has a volumetric Swelling capacity, as In another preferred embodiment, the grafted polymeric measured by the magnitude of the difference between the material corresponds to Formula 1, comprises Zwitterionic global average dry thickness of the grafted polymer layer as pendant groups, the Substrate surface and the grafted poly determined by standard Scanning electron microscopy (SEM) meric material, in combination, constitute a modified surface, 40 and the global average humidified thickness of the grafted and the modified surface exhibits a static contact angle of less polymer layer as determined by environmental scanning elec than 40 degrees. In one such preferred embodiment, the tron microscopy (ESEM), that is less than 200% of the global grafted polymeric material corresponds to Formula 3 and average dry thickness. In one such preferred embodiment, the comprises Sulfobetaine or carboxybetaine pendant groups. In grafted polymeric material corresponds to Formula 3 and one Such preferred embodiment, the grafted polymeric mate 45 comprises Sulfobetaine or carboxybetaine pendant groups. In rial comprises repeat units derived from sulfobetaine meth one Such preferred embodiment, the grafted polymeric mate acrylate, Sulfobetaine acrylate, Sulfobetaine acrylamide, Sul rial comprises repeat units derived from sulfobetaine meth fobetaine methacrylamide, carboxybetaine methacrylate, acrylate, Sulfobetaine acrylate, Sulfobetaine acrylamide, Sul carboxybetaine acrylate, carboxybetaine acrylamide, or car fobetaine methacrylamide, carboxybetaine methacrylate, boxybetaine methacrylamide monomers. In one such pre 50 carboxybetaine acrylate, carboxybetaine acrylamide, or car ferred embodiment, polymeric material is a homopolymer of boxybetaine methacrylamide monomers. In one such pre sulfobetaine methacrylate, sulfobetaine acrylate, sulfobe ferred embodiment, polymeric material is a homopolymer of taine acrylamide, Sulfobetaine methacrylamide, carboxybe sulfobetaine methacrylate, sulfobetaine acrylate, sulfobe taine methacrylate, carboxybetaine acrylate, carboxybetaine taine acrylamide, Sulfobetaine methacrylamide, carboxybe acrylamide, or carboxybetaine methacrylamide monomers 55 taine methacrylate, carboxybetaine acrylate, carboxybetaine and the modified surface exhibits a static contact angle of less acrylamide, or carboxybetaine methacrylamide monomers than 25 degrees. In one such preferred embodiment, the and the grafted polymer layer has a Volumetric Swelling grafted polymeric material is a copolymer, at least 50% of the capacity, as measured by the magnitude of the difference monomeric residues of which are residues of sulfobetaine between the global average dry thickness of the grafted poly methacrylate, Sulfobetaine acrylate, Sulfobetaine acrylamide, 60 mer layer as determined by standard Scanning electron Sulfobetaine methacrylamide, carboxybetaine methacrylate, microscopy (SEM) and the global average humidified thick carboxybetaine acrylate, carboxybetaine acrylamide, or car ness of the grafted polymer layer as determined by environ boxybetaine methacrylamide and the modified surface exhib mental scanning electron microscopy (ESEM), that is less its a static contact angle of less than 25 degrees. In one Such than 200% of the global average dry thickness. In one such preferred embodiment, the grafted polymeric material is a 65 preferred embodiment, the grafted polymeric material is a copolymer, at least 60% of the monomeric residues of which copolymer, at least 50% of the monomeric residues of which are residues of sulfobetaine methacrylate, sulfobetaine acry are residues of sulfobetaine methacrylate, sulfobetaine acry US 9,096,703 B2 63 64 late, Sulfobetaine acrylamide, Sulfobetaine methacrylamide, further example, in each of the foregoing embodiments, the carboxybetaine methacrylate, carboxybetaine acrylate, car grafted polymer layer has a volumetric Swelling capacity that boxybetaine acrylamide, or carboxybetaine methacrylamide may be less than 200%, e.g., less than 100%, less than 50%, and the grafted polymer layer has a Volumetric Swelling less than 25%, less than 10%, less than 5%, less than 1%, or capacity measured by the magnitude of the difference even 0, as measured by the magnitude of the difference between the global average dry thickness of the grafted poly between the global average dry thickness of the grafted poly mer layer as determined by standard Scanning electron mer layer as determined by standard Scanning electron microscopy (SEM) and the global average humidified thick microscopy (SEM) and the global average humidified thick ness of the grafted polymer layer as determined by environ ness of the grafted polymer layer as determined by environ mental scanning electron microscopy (ESEM), that is less 10 mental scanning electron microscopy (ESEM). than 200% of the global average dry thickness. In one such In another preferred embodiment, the grafted polymeric preferred embodiment, the grafted polymeric material is a material corresponds to Formula 1, comprises Zwitterionic copolymer, at least 60% of the monomeric residues of which pendant groups, the Substrate surface and the grafted poly are residues of sulfobetaine methacrylate, sulfobetaine acry meric material, in combination, constitute a modified surface, late, Sulfobetaine acrylamide, Sulfobetaine methacrylamide, 15 and the modified surface exhibits a relatively low affinity for carboxybetaine methacrylate, carboxybetaine acrylate, car proteins. For example, the modified surface may exhibit a boxybetaine acrylamide, or carboxybetaine methacrylamide fibrinogen adsorption of less than 125 ng/cm in a fibrinogen and the grafted polymer layer has a Volumetric Swelling adsorption assay. By way of further example, in one embodi capacity measured by the magnitude of the difference ment the modified Surface may exhibit a fibrinogen adsorp between the global average dry thickness of the grafted poly tion of less than 90 ng/cm in a fibrinogen adsorption assay. mer layer as determined by standard Scanning electron By way of further example, in one embodiment the modified microscopy (SEM) and the global average humidified thick surface may exhibit a fibrinogen adsorption of less than 70 ness of the grafted polymer layer as determined by environ ng/cm in a fibrinogen adsorption assay. By way of further mental scanning electron microscopy (ESEM), that is less example, it is generally preferred that the modified surface than 200% of the global average dry thickness. In one such 25 exhibit a fibrinogen adsorption of less than 50 ng/cm in a preferred embodiment, the grafted polymeric material is a fibrinogen adsorption assay. In one such preferred embodi copolymer, at least 70% of the monomeric residues of which ment, the grafted polymeric material corresponds to Formula are residues of sulfobetaine methacrylate, sulfobetaine acry 3 and comprises Sulfobetaine or carboxybetaine pendant late, Sulfobetaine acrylamide, Sulfobetaine methacrylamide, groups. In one such preferred embodiment, the grafted poly carboxybetaine methacrylate, carboxybetaine acrylate, car 30 meric material comprises repeat units derived from sulfobe boxybetaine acrylamide, or carboxybetaine methacrylamide taine methacrylate, Sulfobetaine acrylate, Sulfobetaine acry and the grafted polymer layer has a volumetric swelling lamide, sulfobetaine methacrylamide, carboxybetaine capacity measured by the magnitude of the difference methacrylate, carboxybetaine acrylate, carboxybetaine acry between the global average dry thickness of the grafted poly lamide, or carboxybetaine methacrylamide monomers. In one mer layer as determined by standard Scanning electron 35 Such preferred embodiment, polymeric material is a microscopy (SEM) and the global average humidified thick homopolymer of sulfobetaine methacrylate, sulfobetaine ness of the grafted polymer layer as determined by environ acrylate, Sulfobetaine acrylamide, Sulfobetaine methacryla mental scanning electron microscopy (ESEM), that is less mide, carboxybetaine methacrylate, carboxybetaine acrylate, than 200% of the global average dry thickness. In one such carboxybetaine acrylamide, or carboxybetaine methacryla preferred embodiment, the grafted polymeric material is a 40 mide monomers and the modified surface exhibits a fibrino copolymer, at least 80% of the monomeric residues of which gen adsorption of less than 30 ng/cm. In one such preferred are residues of sulfobetaine methacrylate, sulfobetaine acry embodiment, the grafted polymeric material is a copolymer, late, Sulfobetaine acrylamide, Sulfobetaine methacrylamide, at least 50% of the monomeric residues of which are residues carboxybetaine methacrylate, carboxybetaine acrylate, car of sulfobetaine methacrylate, sulfobetaine acrylate, sulfobe boxybetaine acrylamide, or carboxybetaine methacrylamide 45 taine acrylamide, Sulfobetaine methacrylamide, carboxybe and the grafted polymer layer has a Volumetric Swelling taine methacrylate, carboxybetaine acrylate, carboxybetaine capacity measured by the magnitude of the difference acrylamide, or carboxybetaine methacrylamide and the between the global average dry thickness of the grafted poly modified surface exhibits a fibrinogen adsorption of less than mer layer as determined by standard Scanning electron 30 ng/cm. In one such preferred embodiment, the grafted microscopy (SEM) and the global average humidified thick 50 polymeric material is a copolymer, at least 60% of the mono ness of the grafted polymer layer as determined by environ meric residues of which are residues of sulfobetaine meth mental scanning electron microscopy (ESEM), that is less acrylate, Sulfobetaine acrylate, Sulfobetaine acrylamide, Sul than 200% of the global average dry thickness. In one such fobetaine methacrylamide, carboxybetaine methacrylate, preferred embodiment, the grafted polymeric material is a carboxybetaine acrylate, carboxybetaine acrylamide, or car copolymer, at least 90% of the monomeric residues of which 55 boxybetaine methacrylamide and the modified surface exhib are residues of sulfobetaine methacrylate, sulfobetaine acry its a fibrinogen adsorption of less than 30 ng/cm. In one such late, Sulfobetaine acrylamide, Sulfobetaine methacrylamide, preferred embodiment, the grafted polymeric material is a carboxybetaine methacrylate, carboxybetaine acrylate, car copolymer, at least 70% of the monomeric residues of which boxybetaine acrylamide, or carboxybetaine methacrylamide are residues of sulfobetaine methacrylate, sulfobetaine acry and the grafted polymer layer has a Volumetric Swelling 60 late, Sulfobetaine acrylamide, Sulfobetaine methacrylamide, capacity measured by the magnitude of the difference carboxybetaine methacrylate, carboxybetaine acrylate, car between the global average dry thickness of the grafted poly boxybetaine acrylamide, or carboxybetaine methacrylamide mer layer as determined by standard Scanning electron and the modified surface exhibits a fibrinogen adsorption of microscopy (SEM) and the global average humidified thick less than 30 ng/cm. In one such preferred embodiment, the ness of the grafted polymer layer as determined by environ 65 grafted polymeric material is a copolymer, at least 80% of the mental scanning electron microscopy (ESEM), that is less monomeric residues of which are residues of sulfobetaine than 200% of the global average dry thickness. By way of methacrylate, Sulfobetaine acrylate, Sulfobetaine acrylamide, US 9,096,703 B2 65 66 Sulfobetaine methacrylamide, carboxybetaine methacrylate, agents is greater than either of the agents alone. A substance, carboxybetaine acrylate, carboxybetaine acrylamide, or car material or agent that is not considered active, can become boxybetaine methacrylamide and the modified surface exhib active if an active agent is immobilized on the Substance, its a fibrinogen adsorption of less than 30 ng/cm. In one such material or agent. Active agents include, but are not limited to preferred embodiment, the grafted polymeric material is a inorganic compounds, organometallic compounds, organic copolymer, at least 90% of the monomeric residues of which compounds or any synthetic or natural, chemical or biological are residues of sulfobetaine methacrylate, sulfobetaine acry compounds of known or unknown therapeutic effect. late, Sulfobetaine acrylamide, Sulfobetaine methacrylamide, In general, a bioactive agent can be immobilized covalently carboxybetaine methacrylate, carboxybetaine acrylate, car or non-covalently directly on the Substrate, on the undercoat boxybetaine acrylamide, or carboxybetaine methacrylamide 10 ing layer, on the grafted polymer layer, or combinations and the modified surface exhibits a fibrinogen adsorption of thereof. In one embodiment, the bioactive agent is immobi less than 30 ng/cm. By way of further example, in each of the lized covalently by reacting one or more functional groups on foregoing embodiments, the modified Surface exhibits a the active agent with one or more functional groups on the fibrinogen adsorption that may be less than 20 ng/cm, e.g., Substrate, undercoating layer, and/or grafted polymer layer. less than 15 ng/cm, less than 12 ng/cm, less than less than 15 Covalent bonds can beformed by a variety of reaction mecha 10, less than 8 ng/cm, less than 6 ng/cm, less than 4, less nisms including, but not limited to, Substitution, addition, and than 2ng/cm, less than 1 ng/cm, less than 0.5 ng/cm, or less condensation reactions. than less than 0.25 ng/cm. Typically, the bioactive agent will typically be immobi Fluorescent and Colorimetric Labels lized on the grafted polymer layer after the grafted polymer In one embodiment, the substrate surface and/or the grafted layer has been grown from the Surface. In an alternative polymer layer is stained or labeled with one or more colori embodiment, the bioactive agent can be co-immobilized with metric labels, fluorescence labels, or combinations thereof. the grafted polymer layer in a side by side structure. In the These labels are used to visualize the surface using the naked graft-from methods, a tether can be grown from the Surface eye, spectroscopy, microscopy, or combinations thereof. and the active agent immobilized on the tether. Alternatively, Suitable microscopy techniques include, but are not limited 25 the active agent can be immobilized directly on the surface to, optical microscopy, fluorescent microscopy, and combi without the use of a tether. nations thereof. Cell adhesion agents can be immobilized to the composi The Surface can be stained through a chemical reaction or tions described herein. The efficacy of a cell adhesion agent in by physical adsorption Such as charge-charge interactions, binding cells in complex environments may be enhanced by hydrophobic interactions, or hydrophilic interactions. Label 30 reducing non-specific protein adsorption on the Surface from ing compounds include, but are not limited to, compounds or which they are presented, given that cell attachment may be a derivatives of rhodamine, fluorescein, coumarin, orange B. competitive process with other protein adsorption. Further, crystal violets, toluidine blue, methyl violet, nuclear fast red, there may an advantage to resisting attachment of any cells methylene blue, malachite green, magenta, acriflavine, and other than those specifically targeted by the cell adhesion other azo compounds. 35 agent to prevent competitive blocking of the Surface. In another embodiment the grafted polymer, such as a Examples of desirable cell attachment agents include, but Zwitterionic polymer, is labeled by incorporating one or more are not limited to, integrin binders. Exemplary integrin bind reactive labeling monomers into the polymer backbone dur ers include, but are not limited to, RGD peptides, along with ing polymerization. These labeling monomers include, but a number of variants that include RGD motifs, YIGSR pep not limited to, FITC-methacrylate, FITC-acrylate, 40 tides, fibronectin, laminin or other proteins or peptides. rhodamine-methacrylate, rhodamine-acrylate, their deriva Longer variants of these peptide may have more specific tives or any other fluorescent acrylate, methacrylate, acryla target cell binding. Further, the ability to present locally dense mide, vinyl compound, diol or diamine. Incorporation of concentrations of cell attachment agents may increase the these groups can allow for convenient measurement of con effectiveness of cell attachment by creating multimeric inter formality and/or grafted polymer layer thickness. This may 45 actions. Other cell adhesion agents include, but are not lim be particularly useful as a quality control metric for confor ited, to REDV peptides. Tailored integrin binders can be used mality verification during manufacturing of the grafted poly for a variety of applications including osteointegration. mer layer on an underlying device. Cell adhesion agents that bind specific immune cells may In another embodiment, the grafted polymer layer is also benefit from attachment to Zwitterions. Adhesion of stained with one or more compounds, which can be easily 50 immune cells to the biomaterial surface activates these cells visualized under an electronic microscope (SEM or TEM). and prefaces their phenotypic response, Such as the transition These compounds include, but are not limited to osmium of monocytes to macrophages that can result, in Some cases, tetroxide and ruthenium tetroxide. in the fusion into undesirable foreign body giant cells. The Bioactive Agents inherent resistivity to random protein fouling that Zwitterions Therapeutics, diagnostic, and/or prophylactic agents can 55 possess provides a unique platform to couple biomolecules be immobilized on or otherwise incorporated into an article of that act as specific ligands for immune cells including neu the present invention. When optionally included, such bioac trophils, monocytes, helper T-cells, killer T-cells, suppressor tive agents may be leachable or non-leachable. For example, T-cells, B-cells and dendritic cells. Selection of appropriate the bioactive agent may be dissolved or otherwise contained ligands may prime these cells for beneficial instead of detri within the Substrate, or covalently or non-covalently associ 60 mental functions. These ligands include peptides or proteins ated with the grafted polymer layer, and leached or otherwise that specifically bind immune cell receptors such as integrins, disassociated with the article in a controlled or uncontrolled selectins, complement, or Fc gamma. When bound to these manner (e.g., by leaching). These agents can interact pas cell-associated proteins, such ligands may stimulate intracel sively or actively with the Surrounding in Vivo environment. lular signaling pathways that lead to responses including The agents can also be used to alter the Surrounding in vivo 65 cytoskeletal rearrangements, production and secretion of chemistry or environment. Two or more agents can be immo molecules including chemokines, cytokines and other bilized to a substrate surface, wherein the activity of the two chemoattractants, and induction of apoptosis. Desirable US 9,096,703 B2 67 68 behaviors that could be tailored by presentation of biomol cysteine residue which can be used to immobilize the peptide ecules via Zwitterionic tethers may include prevention/reduc on a Surface by reacting the thiol group of the cysteine residue tion in the secretion of proinflammatory cytokines, enhance with a thiol-reactive group on the Surface. ment of phagocytosis, and modulation of the release of Tethering of these agents via non-fouling materials, such as soluble factors that influence tissue-device integration. 5 Zwitterions, should provide stable, long-term activity. Addi Osteointegration may also be promoted or induced by fac tionally, immobilization of enzymes that degrade bacterial tors which would benefit from the non-fouling properties and attachment and biofilm proteins, such as glycosylases, lyases, stable presentation of non-fouling materials, such as Zwitte and serine-proteases, or those that degrade microbial com rions. Osteointegration promoting agents include, but are not munication signal molecules, such as N-acyl-homoserine lac limited to, bone-morphogenic proteins, such as BMP2 and 10 tone acylases, could provide improved efficacy in prevention shortened analogues thereof. Non-fouling Surfaces, such as of initial microbial adhesion events and subsequent biofilm Zwitterionic Surfaces, may enhance the activity of agents formation. designed to promote desired cell regrowth over a surface. A broad range of antimicrobial or antiseptic agents may be Reducing attachment of neutrophils and macrophages may incorporated in the Substrate or the non-fouling polymer to inhibit the foreign body response and enable desired cell 15 enhance antimicrobial activity at the surface or be released to attachment and growth process to be favored. provide antimicrobial activity in the environment Surround Presentation of antithrombotic agents may also be more ing the article. Suitable agents include silver metals, silver effective when tethered to grafted polymers, such as Zwitte salts such as silver Sulfadiazine, silver oxide, silver carbonate, rionic materials, relative to other tethers. The process of silver acetate, silveralginate, silver azide, silver citrate, silver thrombosis involves both surface and bulk pathways. Zwitte lactate, silver nitrate, silver sulfate, silver chloride, silver rions have shown an ability to reduce platelet attachment and thiocyanate, silver-sodium-hydrogen-Zirconium phosphate, activation, reducing one pathway. Combining an active anti silver Sulfadiazine, silver cyclohexanediacetic acid and disil thrombotic that assists in the reduction of platelet activation ver 2,5-dichloro-3,6-dihydroxy-2,5-cyclohexadiene-1,4-di or directly targets additional pathways for thrombosis with a one, among others, a bismuth Salt Such as bismuth nitrate, Zwitterionic tether could enhance the antithrombotic effect 25 bismuth citrate or bismuth Salicylate among others, a Zinc compared to either a non-platelet adherent Surface or the salt, a cerium salt, triclosan, combinations of chlorhexidine antithrombotic agent alone. Suitable antithrombotic agents free base and chlorhexidine acetate, benzalkonium chloride, include, but are not limited to, thrombomodulin, heparin, citrate, povidoneiodine, parachlorometaxylene, gramicidin, heparin fragments, derivatized heparin fragments, hyaluronic polymixin, norfloxacin, tobramycin, Sulfamylon, polyhex acid, reversible albumin binders, tissue plasminogen activa 30 amethylene biguanide, alexidine, iodine, rifampicin, micona tor binders, transglutimase, reversible NO binders, polyl Zole, bacitracin, and minocycline, ciprofloxacin, clindamy ysine, sulphonated polymers, thrombin inhibitors including cin, erythromycin, gentamycin, tetracycline and Vancomycin. hirudin, urokinase, and streptokinase. Biguanide compounds which may be used according to the Device-centered infection remains a large problem. Non invention include poly(hexamethylene biguanide) hydro fouling materials, such as Zwitterions materials, can by them 35 chloride and chlorhexidine compounds. Chlorhexidine is the selves diminish microbial adhesion and retard biofilm devel term denoting the chemical compound 1.6 bis(N5-p-chlo opment. Prevention of microbial adhesion and biofilm can be rophenyl-N1-biguanido)hexane). Chlorhexidine compounds further enhanced on non-fouling Surfaces, such as Zwitteri include chlorhexidine free base (“CHX”) as well as chlo onic Surfaces, by presentation of antimicrobials including, rhexidine salts, such as chlorhexidine diphosphanilate, chlo but not limited to, membrane-targeting antimicrobial agents, 40 rhexidine digluconate (“CHG'), chlorhexidine diacetate antimicrobial peptides and Small molecule antimicrobial (“CHA), chlorhexidine dihydrochloride, chlorhexidine agents. Generally, antimicrobial peptides are cationic mol dichloride, chlorhexidine dihydroiodide, chlorhexidine ecules with spatially separated hydrophobic and charged diperchlorate, chlorhexidine dinitrate, chlorhexidine sulfate, regions. Exemplary antimicrobial peptides include linear chlorhexidine sulfite, chlorhexidine thiosulfate, chlorhexi peptides that form an O.-helical structure in membranes or 45 dine di-acid phosphate, chlorhexidine difluorophosphate, peptides that form B-sheet structures, optionally stabilized chlorhexidine diformate, chlorhexidine dipropionate, chlo with disulfide bridges in membranes. Representative antimi rhexidine di-iodobutyrate, chlorhexidine di-n-Valerate, chlo crobial peptides include, but are not limited to, cathelicidins, rhexidine dicaproate, chlorhexidine malonate, chlorhexidine defensins, dermcidin, and more specifically magainin 2, pro Succinate, chlorhexidine malate, chlorhexidine tartrate, chlo tegrin, protegrin-1, melittin, II-37, dermaseptin 01, cecropin, 50 rhexidine dimonoglycolate, chlorhexidine mono-diglycolate, caerin, ovispirin, cecropin Amelittin hybrid, andalamethicin, chlorhexidine dilactate, chlorhexidine di-O-hydroxyisobu or hybrids or analogues of other AmPs. Naturally occurring tyrate, chlorhexidine diglucoheptonate, chlorhexidine antimicrobial peptides include peptides from vertebrates and di-isothionate, chlorhexidine dibenzoate, chlorhexidine non-vertebrates, including plants, humans, fungi, microbes, dicinnamate, chlorhexidine dimandelate, chlorhexidine di and insects. 55 isophthalate, chlorhexidine di-2-hydroxy-napthoate, and Antimicrobial peptides can be made from naturally occur chlorhexidine embonate. ring amino acids, non-naturally occurring amino acids (e.g., Bismuth salts which may be used according to the inven synthetic or semisynthetic amino acids and peptidomimet tion include bismuth nitrate, bismuth citrate, bismuth salicy ics), or combinations thereof. Antimicrobial peptides which late, bismuth borate, bismuth mandelate, bismuth palmitate, retain their activity when immobilized on a Surface are gen 60 bismuth benzoate, and bismuth sulfadiazine. erally referred to as membrane-targeting antimicrobial Cerium salts which may be used according to the invention agents. Antimicrobial peptides can be immobilized on the include cerium nitrate and other cerium salts having a water non-fouling grafted polymer layer, the Substrate, the under solubility similar to cerium nitrate. coating or combinations thereof by reacting a functional The term silver-containing compound, as used herein, group on the peptide with a functional group on the non 65 refers to a compound comprising silver, either in the form of fouling grafted polymer layer, the Substrate, and/or the primer a silver atom or a silver ion unlinked or linked to another coat. For example, the peptide can be designed to have a molecule via a covalent or noncovalent (e.g., ionic) linkage, US 9,096,703 B2 69 70 including but not limited to covalent compounds such as dilaZep hydrochloride, Verapamil, nicardipine, nicar silver sulfadiazine (AgSD) and silver salts such as silver dipine hydrochloride and Verapamil hydrochloride) oxide (AgO), silver carbonate (AgCO), silver deoxy Peripheral vasodilators (such as ifenprodil tartrate, cholate, silver salicylate, silver iodide, silver nitrate cinepacide maleate, ciclandelate, cynnaridine and pen ('AgNO), silver paraminobenzoate, silver paraminosalicy toxyphylin) late, silver acetylsalicylate, silver ethylenediaminetetraacetic Antibiotics (such as amplicillin, amoxicillin, cefalexin, acid ('Ag EDTA), silver picrate, silver protein, silver citrate, cephalexin, cefoxytin and cephalothin, erythromycm silver lactate and silver laurate. ethyl Succinate, vacampicillin hydrochloride, minocy Zinc salts which may be used according to the invention cline hydrochloride, chloramphenicol, tetracycline, include Zinc acetate and other Zinc salts having a water Solu 10 erythromycin, ceftazidime, cefuroxime Sodium, bility similar to Zinc acetate. aspoxicillin chloramphenicol, clindamycin, erythromy The classes of bioactive agents identified above may be cin, erythromycin ethyl carbonate, erythromycin esto incorporated in the Substrate or the non-fouling polymer to late, erythromycin glucepate, erythromycin ethylsucci enhance antimicrobial activity at the surface or be released to 15 nate, erythromycin lactobionate, roXithromycin, provide antimicrobial activity in the environment Surround lincomycin, natamycin, nitrofurantoin, spectinomycin, ing the article. Vancomycin, aztreonarn, colistin IV, metronidazole, Additional groups/classes of bioactive agents may be tinidazole, fusidic acid, trimethoprim, and 2-thiopyri incorporated in the Substrate or the non-fouling polymer to dine N-oxide) enhance antimicrobial activity at the surface or be released to Synthetic antimicrobials (such as nalidixic acid, piromidic provide antimicrobial activity in the environment Surround acid, pipemidic acid trihydrate, enoxacin, cinoxacin, of ing the article and include the following groups/classes: loxacin, norfloxacin, ciprofloxacin hydrochloride and Antipyretics, analgesics and antiphlogistics (Such as sulfamethoxazole-trimethoprim) indometacin, acetylsalicylic acid, diclofenac sodium, Antiviral agents (such as acyclovir, ganciclovir, acyclovir ketoprofen, ibuprofen, mefenamic acid, aZulene, phen 25 prodrugs, famcyclovir, Zidovudine, didanosine, stavu acetin, isopropyl antipyrine, acetaminophen, benzadac, dine, lamivudine, Zalcitabine, Saquinavir, indinavir, phenylbutaZone, flufenamic acid, acetylsalicylic acid ritonavir, n-docosanol, tromantadine and idoxuridine) (aspirin), paracetamol, phenaZone, Sodium salicylate, Anticonvulsants (such as propantheline bromide, atropine Salicylamide, Sazapyrine, and etodolac) Opioid analge Sulfate, oxitropium bromide, timepidium bromide, Sco sics (such as buprenorphine, dextromoramide, dextro 30 polamine butylbromide, troSpium chloride, butropium propoxyphene, fentanyl, alfentanil, Sufentanil, hydro bromide, N-methylscopolaminemethylsulfate and morphone, methadone, morphine, oxycodone, methyloctatropine bromide) papavereturn, pentazocine, pethidine, phenopefidine, Antitussives (such as tipepedine hibenzate, methylephe codeine dihydrocodeine) Non-selective COX inhibitors drine hydrochloride, codeine phosphate, tranilast, dex Such as Salicylic acid derivatives, aspirin, Sodium sali 35 tromethorphan hydrobromide, dimemorfan phosphate, cylate, choline magnesium trisalicylate, Salsalate, clobutinol hydrochloride, fominoben hydrochloride, diflunisal, SulfaSalazine and olSalazine). Para-ami benproperine phosphate, eprazinone hydrochloride, nophenol derivatives Such as acetaminophen. Indole and clofedanol hydrochloride, ephedrine hydrochloride, indene acetic acids such as indomethacin and Sulindac. noscapine, pentoxyverine citrate, oxeladin citrate and Heteroaryl acetic acids such as tolmetin, dicofenac and 40 isoaminyl citrate) ketorolac. Arylpropionic acids such as ibuprofen, Expectorants (such as bromhexine hydrochloride, car naproxen, flurbiprofen, ketoprofen, fenoprofen and bocysteine, ethylcysteine hydrochloride and methylcys oxaprozin. Anthranilic acids (fenamates) Such as mefe teine hydrochloride) namic acid and meloxicam. Enolic acids such as the Bronchodilators (such as theophylline, aminophylline, oxicams (piroxicam, meloxicam). Alkanones Such as 45 Sodium cromoglicate, procaterol hydrochloride, trime nabumetone. Selective COX-2 Inhibitors (such as dia toquinol hydrochloride, diprophilline, salbutamol Sul ryl-substituted furanones such as rofecoxib; diaryl-sub fate, clorprenaline hydrochloride, formoterol fumarate, stituted pyrazoles such as celecoxib; indole acetic acids ocriprenaline sulfate, pilbuterol hydrochloride, hexo Such as etodolac and Sulfonanilides Such as nimeSulide) prenaline sulfate, bitolterol mesilate, clenbuterol hydro Anti-inflammatory steroids (such as cortisone, hydrocorti 50 chloride, terbutaline sulfate, malbuterol hydrochloride, Sone, prednisone, dexamethasone, methylprednisolone, fenoterol hydrobromide and methoxyphenamine hydro triamcinolone beclomethasone flunisolide, fluticasone chloride), (13) cardiotonics (such as dopamine hydro proprionate triamcinolone acetonide budesonide lot chloride, dobutamine hydrochloride, docarpamine, erednoletabonate and mometasone, aclometasone, des denopamine, caffeine, digoxin, digitoxin and ubide onide, hydrocortisone, betamethasone, clocortolone, 55 carenone) desoximetaSone, fluocinolone, flurandrenolide, Diuretics (such as furosemide, acetazolamide, triclorme mometasone, prednicarbate; amcinonide, desoximeta thiazide, methylclothiazide, hydrochlorothiazide, Sone, diflorasone, fluocinolone, fluocinonide, halcinon hydroflumethiazide, ethiazide, cyclopenthiazide, ide, clobetasol, augmented betamethasone, diflorasone, spironolactone, triamterene, florothiazide, piretanide, halobetasol, prednisone, dexamethasone and methyl 60 mefruside, etacrynic acid, azosemide and clofenamide) prednisolone and their derivatives and) Muscle relaxants (such as chlorphenesin carbamate, tolp Antiulcer drugs (such as ecabet sodium, emprostil, erisone hydrochloride, eperisone hydrochloride, tizani Sulpiride, cetraxate hydrochloride, gefarnate, irsoglad dine hydrochloride, mefenicine, chlorZoxaZone, phen ine maleate, cimetidine, ranitidine hydrochloride, famo probamate, methocarbamol, chlormeZaZone, pridinol tidine, nizatidine and roXatidine acetate hydrochloride) 65 mesilate, afloqualone, baclofen and dantrolene Sodium) Coronary vasodilators (such as nifedipine, isosorbide dini Cerebral metabolism ameliorants (such as nicergoline, trate, diltiazem hydrochloride, trapidil, dipyridamole, meclofenoxate hydrochloride and taltirelin), US 9,096,703 B2 71 72 Minor tranquilizers (such as oxazolam, diazepam, clotiaz applicable (such as ibuprofen, flurbiprofen, ketoprofen, epam, medazepam, temazepam, fludiazepam, mep aclofenac, diclofenac, aloxiprin, aproxen, aspirin, robamate, nitrazepam and chlordiazepoxide) diflunisal, fenoprofen, indomethacin, mefenamic acid, Major tranquilizers (such as Sulpiride, clocapramine naproxen, phenylbutaZone, piroxicam, salicylamide, hydrochloride, Zotepine, chlorpromazine and haloperi Salicylic acid, Sulindac, desoxysulindac, tenoxicam, tra dol) madol, ketoralac, flufenisal, Salsalate, triethanolamine Beta-blockers (such as bisoprolol fumarate, pindolol, pro Salicylate, aminopyrine, antipyrine, oxyphenbutaZone, pranolol hydrochloride, carteolol hydrochloride, meto apaZone, cintaZone, flufenamic acid, clonixerl, clonixin, prolol tartrate, labetanol hydrochloride, acebutolol meclofenamic acid, flunixin, coichicine, demecolcine, hydrochloride, bufetolol hydrochloride, alprenolol 10 allopurinol, oxypurinol, benzydamine hydrochloride, hydrochloride, arotinolol hydrochloride, oxprenolol dimefadane, indoxole, intrazole, mimbane hydrochlo hydrochloride, nadolol, bucumorolhydrochloride, inde ride, paranylene hydrochloride, tetrydamine, benzin nolol hydrochloride, timolol maleate, befunolol hydro dopyrine hydrochloride, fluprofen, ibufenac, naproXol, chloride and bupranolol hydrochloride) fenbufen, cinchophen, diflumidone sodium, fenamole, Antiarrthymics (such as procainamide hydrochloride, 15 flutiazin, metazamide, letimide hydrochloride, nexeri diso-pyramide, ajmaline, quinidine Sulfate, aprindine dine hydrochloride, octaZamide, molinazole, neocin hydrochloride, propafenone hydrochloride, mexiletine chophen, nimazole, proxazole citrate, tesicam, tesimide, hydrochloride and azmilide hydrochloride) tolimetin, and triflumidate) Athrifuges (such as allopurinol, probenicid, colchicine, Antineoplastic/antiangiogenic (Such as acivicin, aclarubi Sulfinpyrazone, benzbromarone and bucolome) cin, acodazole, acronycine, adoZelesin, alanosine, Anticoagulants/Antiplatelets (such as heparin, chondroi aldesleukin, allopurinol sodium, altretamine, aminoglu ten sulfate ticlopidine hydrochloride, dicumarol, potas tethimide, amonafide, ampligen, amsacrine, androgens, sium warfarin, and (2R,3R)-3-acetoxy-5-2-(dimethy anguidine, aphidicolin glycinate, asaley, asparaginase, lamino)ethyl-2,3-dihydro-8-methyl-2-(4-me 5-azacitidine, azathioprine, Bacillus calmette-guerin thylphenyl)-1,5-benzothiazepin-4(5H)-onemaleate) 25 (BCG), Baker's Antifol (soluble), beta-2'-deox Thrombolytics (such as stretokinase, urokinase and tissue ythioguanosine, bisantrene hcl, bleomycin Sulfate, plasminogin activators, methyl (2E.3Z)-3-benzylidene busulfan, buthionine sulfoximine, BWA 773U82, BW 4-(3,5-dimethoxy-C.-methylbenzylidene)-N-(4-meth 502U83.HCl, BW 7U85 mesylate, ceracemide, carbe ylpiperazin-1-yl)-Succinamate hydrochloride), timer, carboplatin, carmustine, chlorambucil, chloroqui Liver disease drugs (such as (+)r-5-hydroxymethyl-t-7-(3, 30 noxaline-Sulfonamide, chlorozotocin, chromomycin 4-dimethoxyphenyl)-4-oxo-4,5,6,7-tetrahydrobenzob. A3, cisplatin, cladribine, corticosteroids, Corynebacte furan-c-6-carboxylactone) rium parvum, CPT-11, crisinatol, cyclocytidine, cyclo Antiepileptics (such as phenyloin, sodium valproate, met phosphamide, cytarabine, cytemberna, dabis maleate, albital and carbamazepine) dacarbazine, dactinomycin, daunorubicin HCl, deazau Antihistamines (such as chlorpheniramine maleate, clem 35 ridine, dexraZoxane, dianhydrogalactitol, diaziquone, astine fumarate, meduitazine, alimemazine tartrate, dibromodulcitol, didemnin B, diethyldithiocarbamate, cyproheptadine hydrochloride and bepotastin besilate) diglycoaldehyde, dihydro-5-azacytidine, doxorubicin, Antiemitics (such as difenidol hydrochloride, metoclopra echinomycin, edatrexate, edelfosine, eflornithine, mide, domperidone and betahistine mesilate and trime Elliott's solution, elsamitrucin, epirubicin, esorubicin, butine maleate), 40 estramustine phosphate, estrogens, etanidazole, ethio Depressors (such as dimethylaminoethyl reserpilinate fos, etoposide, fadrazole, fazarabine, fenretinide, dihydrochloride, rescinnamine, methyldopa, prazocin filgrastim, finasteride, flavone acetic acid, floXuridine, hydrochloride, bunazosin hydrochloride, clonidine fludarabine phosphate, 5-fluorouracil, Fluosol R, fluta hydrochloride, budralazine, urapidil and N-6-2-(5- mide, gallium nitrate, gemcitabine, goserelin acetate, bromo-2-pyrimidinyl)oxyethoxy-5-(4-methylphe 45 hepsulfam, hexamethylene bisacetamide, homohar nyl)-4-pyrimidinyl)-4-(2-hydroxy-1,1-dimethyl-ethyl) ringtonine, hydrazine Sulfate, 4-hydroxyandrostenedi benzenesulfonamide Sodium) one, hydrozyurea, idarubicin HCl, ifosfamide, inter Hyperlipidemia agents (such as pravastatin Sodium and feron alfa, interferon beta, interferon gamma, fluvastatin Sodium) interleukin-1 alpha and beta, interleukin-3, interleukin Sympathetic nervous stimulants (such as dihydroergota 50 4, interleukin-6.4-ipomeanol, iproplatin, isotretinoin, mine mesilate and isoproterenol hydrochloride, etile leucovorin calcium, leuprolide acetate, levamisole, lipo frine hydrochloride) Somal daunorubicin, liposome encapsulated doxorubi Oral diabetes therapeutic drugs (such as glibenclamide, cin, lomustine, lonidamine, maytansine, mechlore tolbutamide and glimidine Sodium) thamine hydrochloride, melphalan, menogaril, Oral carcinostatics (such as malimastat) 55 merbarone, 6-mercaptopurine, mesna, methanol extrac Alkaloid narcotics (such as morphine, codeine and tion residue of Bacillus calmette-guerin, methotrexate, cocaine) N-methylformamide, mifepristone, mitoguaZone, mito Vitamins (such as vitamin B1, vitamin B2, vitamin B6, mycin-C, mitotane, mitoxantrone hydrochloride, mono vitamin B12, vitamin C and folic acid) cyte/macrophage colony-stimulating factor, nabilone, Thamuria therapeutic drugs (such as flavoxate hydrochlo 60 nafoxidine, neocarzinostatin, octreotide acetate, orma ride, oxybutynin hydrochloride and terolidine hydro platin, oxaliplatin, paclitaxel, pala, pentostatin, pipera chloride) Zinedione, pipobroman, pirarubicin, piritrexim, piroX Angiotensin converting enzyme inhibitors (such as imi antrone hydrochloride, PIXY-321, plicamycin, porfimer dapril hydrochloride, enalapril maleate, alacepril and Sodium, prednimustine, procarbazine, progestins, pyra delapril hydrochloride). 65 Zoflurin, razoxane, SargramoStim, Semustine, spiroger Non-steroidal anti-inflammatory agents including their manium, Spiromustine, Streptonigrin, streptozocin, racemic mixtures or individual enantiomers where Sulofenur, Suramin Sodium, tamoxifen, taxotere, tegafur, US 9,096,703 B2 73 74 teniposide, terephthalamidine, teroxirone, thioguanine, (HGF), connective tissue growth factor (CTGF), thiotepa, thymidine injection, tiazofurin, topotecan, myeloid colony-stimulating factors (CSFs), monocyte toremifene, tretinoin, trifluoperazine hydrochloride, tri chemotactic protein, granulocyte-macrophage colony fluridine, trimetrexate, tumor necrosis factor, uracil stimulating factors (GM-CSF), granulocyte colony mustard, vinblastine Sulfate, Vincristine Sulfate, Vin stimulating factor (G-CSF), macrophage colony-stimu desine, vinorelbine, Vinzolidine, Yoshi 864, Zorubicin, lating factor (M-CSF), erythropoietin, interleukins and mixtures thereof) (particularly IL-1, IL-8, and IL-6), tumor necrosis fac Immunosuppressant agents (such as cyclosporine A, tor-O. (TNF9), nerve growth factor (NGF), interferon-C. mycophenolic acid, tacrolimus, rapamycin, rapamycin interferon-B, histamine, endothelin-1, angiotensin II, analogues, azathioprine, recombinant or monoclonal 10 growth hormone (GH), and synthetic peptides, ana antibodies to interleukins, T-cells, B-cells and/or their logues orderivatives of these factors are also suitable for receptors. release from specific implants and devices to be Vasodilators (such as cyclandelate, isoxSuprine, papaver described later. Other examples include CTGF (connec ine, dipyrimadole, isosorbide dinitrate, phentolamine, tive tissue growth factor); inflammatory microcrystals nicotinyl alcohol, co-dergocrine, nicotinic acid, glycerl 15 (e.g., crystalline minerals such as crystalline silicates); trinitrate, pentaerythritol tetranitrate and xanthinol) bromocriptine, methylsergide, methotrexate, chitosan, Antiproliferative agents (such as paclitaxel, actinomycin N-carboxybutyl chitosan, carbon tetrachloride, thioac D., rapamycin, tacrolimus, everolimus, dexamethasone etamide, fibrosin, ethanol, bleomycin, naturally occur and rapamycin analogues) ring or synthetic peptides containing the Arg-Gly-Asp Local anaesthetics (such as benzocaine, bupivacaine, ame (RGD) sequence, generally at one or both termini (see thocaine, lignocaine, lidocaine, cocaine, cinchocaine, e.g., U.S. Pat. No. 5,997.895), and tissue adhesives, such dibucaine, mepivacaine, prilocalne, etidocaine, Veratri as cyanoacrylate and crosslinked poly(ethylene glycol)- dine (specific c-fiber blocker) and procaine) methylated collagen compositions, such as described Antifungals (such as amorolfine, isoconazole, clotrima below. Other examples of fibrosis-inducing agents Zole, econazole, miconazole, nystatin, terbinafine, 25 include bone morphogenic proteins (e.g., BMP-2, BMP bifonazole, amphotericin, griseo fulvin, ketoconazole, 3, BMP-4, BMP-5, BMP-6 (Vgr-1), BMP-7 (OP-1), fluconazole and flucytosine, salicylic acid, fezatione, BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13, ticlatone, tolnaftate, triacetin, Zinc, pyrithione and BMP-14, BMP-15, and BMP-16. Of these, BMP-2, Sodium pyrithione) BMP-3, BMP-4, BMP-5, BMP-6, and BMP-7 are of Agents/chemicals that block microbial attachment to target 30 particular utility. Bone morphogenic proteins are cells and/or inhibits entry of infectious pathogens (e.g., described, for example, in U.S. Pat. Nos. 4,877,864; sulphated and sulponated polymers such as PC-515 (car 5,013,649; 5,661.0075,688,678; 6,177,406; 6.432,919; rageenan), Pro-2000, and Dextrin 2 Sulphate) and 6,534.268 and Wozney, J. M., et al. (1988) Science: Antiretroviral agents (e.g., PMPA gel) that prevent retro 242(4885); 1528 1534. viruses from replicating in the cells 35 Other representative fibrosis-inducing agents include com Agents which change the condition of the tissue to make it ponents of extracellular matrix (e.g., fibronectin, fibrin, hostile to the pathogen (Such as Substances which alter fibrinogen, collagen (e.g., bovine collagen), fibrillar and mucosal pH (e.g., Buffer Gel and Acid form): non-fibrillar collagen, adhesive glycoproteins, pro Agents that treat or prevent an allergic or immune response teoglycans (e.g., heparin Sulfate, chondroitin Sulfate, and/or cellular proliferation (Such as various cytokine 40 dermatan Sulfate), hyaluronan, secreted protein acidic inhibitors such as humanized anti-cytokine antibodies, and rich in cysteine (SPARC), thrombospondins, tena anti-cytokine receptor antibodies, recombinant antago cin, and cell adhesion molecules (including integrins, nists, or soluble receptors; various leucotriene modifiers vitronectin, fibronectin, laminin, hyaluronic acid, elas Such as Zafirlukast, montelukast and Zileuton; immuno tin, bitronectin), proteins found in basement mem globulin E (IgE) inhibitors such as Omalizumab (an 45 branes, and fibrosin) and inhibitors of matrix metallo anti-IgE monoclonal antibody) and secretory leukocyte proteinases, such as TIMPs (tissue inhibitors of matrix protease inhibitor) and SYK Kinase inhibitors) metalloproteinases) and synthetic TIMPs, e.g., mari Agents that prevent restenosis (such as paclitaxel, siroli mistat, batimistat, doxycycline, tetracycline, minocy mus, everolimus, Vincristine, biolimus, mycophenolic cline, TROCADE, Ro-1130830, CGS 27023A, and acid, ABT-578, cervistatin, simvastatin, methylpred 50 BMS-275291. nisolone, dexamethasone, actinomycin-D, angiopeptin, Anti-thrombotic and/or antiplatelet agents (include hep L-arginine, estradiol, 17-B-estradiol, tranilast, methotr arin, heparin fragments, organic salts of heparin, heparin exate, batimistat, halofuginone, BCP-671, QP-2, lant complexes (e.g., benzalkonium heparinate, tridodecy runculin D. cytochalasin A, nitric oxide, and analogues lammonium heparinate, heparin-tridodecylmethylam and derivatives) 55 monium chloride, heparin-benzalkonium chloride, hep Growth factors and inflammatory cytokines involved in arin-steralkonium chloride, heparin-poly-N-vinyl angiogenesis, fibroblast migration, fibroblast prolifera pyrrolidone, heparin-lecithin, heparin tion, ECM synthesis and tissue remodeling, such as epi didodecyldimethylammonium bromide, heparin dermal growth factor (EGF) family, transforming pyridinium chloride, and heparin-synthetic glycolipid growth factor-O. (TGF-C), transforming growth factor-B 60 complex), dextran, Sulfonated carbohydrates such as (TGF-9-1, TGF-9-2, TGF-9-3, platelet-derived growth dextran Sulphate, coumadin, coumarin, heparinoid, dan factor (PDGF), fibroblast growth factor (acidic—aFGF: aparoid, argatroban chitosan Sulfate, chondroitin Sul and basic—bFGF), fibroblast stimulating factor-1, fate, danaparoid, lepirudin, hirudin, AMP adenosine, activins, vascular endothelial growth factor (including 2-chloroadenosine, aspirin, phenylbutaZone, VEGF-2, VEGF-3, VEGF-A, VEGF-B, VEGF-C, pla 65 indomethacin, meclofenamate, hydrochloroquine, dipy cental growth factor—PIGF), angiopoietins, insulin ridamole, iloprost, streptokinase, and factor Xa inhibi like growth factors (IGF), hepatocyte growth factor tors, such as DX9065a, magnesium, and tissue plasmi US 9,096,703 B2 75 76 nogen activator. In one aspect, the anti-thrombotic agent alemtuzumab, gemtuzumab, rituximab, ibritumomab, is a modified heparin compound, Such as a hydrophobi to situmomab, edrecolomab, cetuximab, bevacizumab, cally modified heparin or modified hirudin compound RanibiZumab, Satumomab, pertuZumab, and dacli (e.g., Stearylkonium heparin, benzalkonium heparin, Zumab) cetylkonium heparin, or traodecylmethyl ammonium 5 Therapeutic enzymes (Such as recombinant human tissue heparin). Further examples of anti-thrombotic agents plasminogen activator (alteplase), RNaseA, RNaselJ. include plasminogen, lys-plasminogen, alpha-2-anti chondroitinase, pegaspargase, arginine deaminase, plasmin, urokinase, ticlopidine, clopidogrel, glycopro Vibriolysin, sarcosidase, N-acetylgalactosamine-4-sul tein IIb/IIIa inhibitors such as abcixamab, eptifibatide, fatase, glucocerebrocidase, C.-galactosidase, and and tirogiban. Other agents capable of affecting the rate 10 laronidase) of clotting include glycosaminoglycans, danaparoid, Enzyme inhibitors (Such as edrophonium chloride, N-me 4-hydroxycourmarin, warfarin Sodium, dicumarol, thylphysostigmine, neostigmine bromide, physostig phenprocoumon, indan-1,3-dione, acenocoumarol, mine Sulfate, tacrine HCL, tacrine, 1-hydroxy maleate, anisindione, and rodenticides including bromadiolone, iodotubercidin, p-bromotetramisole, 10-(C-diethylami brodifacoum, diphenadione, chlorophacinone, and pid 15 nopropionyl)-phenothiazine hydrochloride, calmidazo none). lium chloride, hemicholinium-3,3,5-dinitrocatechol, Polypeptide drugs (such as but are not limited to, insulin; diacylglycerol kinase inhibitor I, diacylglycerol kinase growth factors, such as epidermal growth factor (EGF), inhibitor II, 3-phenylpropargylaminie, N-monomethyl insulin-like growth factor (IGF), transforming growth L-arginine acetate, carbidopa, 3-hydroxybenzylhydra factor (TGF), nerve growth factor (NGF), platelet-de zine HCl, hydralazine HCl, clorgyline HCl, deprenyl rived growth factor (PDGF), bone morphogenic protein HCl L(-), deprenyl HCl D(+), hydroxylamine HCl, (BMP), fibroblast growth factor and the like; somatosta iproniazid phosphate, 6-MeO-tetrahydro-9H-pyrido-in tin: Somatotropin; somatropin; Somatrem, calcitonin; dole, nialamide, pargyline HCl, quinacrine HCl, semi parathyroid hormone; colony Stimulating factors (CSF); carbazide HCl, tranylcypromine HCl, N,N-diethylami clotting factors; tumor necrosis factors; interferons; 25 noethyl-2,2-di-phenylvalerate hydrochloride, interleukins; gastrointestinal peptides, such as vasoac 3-isobutyl-1-methylxanthne, papaverine HC1, tive intestinal peptide (VIP), cholecytokinin (CCK), indomethacind, 2-cyclooctyl-2-hydroxyethylamine gastrin, secretin, and the like; erythropoietins; growth hydrochloride, 2,3-dichloro-O-methylbenzylamine hormone and GRF; vasopressins; octreotide; pancreatic (DCMB), 8,9-dichloro-2,3,4,5-tetrahydro-1H-2-benza enzymes; dismutases Such as Superoxide dismutase; thy 30 Zepine hydrochloride, p-aminoglutethimide, p-amino rotropin releasing hormone (TRH); thyroid stimulating glutethimide tartrate R(+), p-aminoglutethimide tartrate hormone; luteinizing hormone: LHRH; GHRH: tissue S(-), 3-iodotyrosine, alpha-methyltyrosine L(-), alpha plasminogen activators; macrophage activator, chori methyltyrosine D(-), cetazolamide, dichlorphenamide, onic gonadotropin; heparin; atrial natriuretic peptide; 6-hydroxy-2-benzothiazolesulfonamide, and allopu hemoglobin, retroviral vectors; relaxin; cyclosporin; 35 rinol) oxytocin, and peptide or polypeptide vaccines. Cell Steroids (Such as glucocorticoids, estrogens and andro response modifiers. (Cell response modifiers include gens. By way of example, Steroids can include dexam chemotactic factors such as platelet-derived growth fac ethasone, dexamethasone acetate, dexamethasone tor (PDGF), pigmented epithelium-derived factor Sodium phosphate, cortisone, cortisone acetate, hydro (PEDF), neutrophil-activating protein, monocyte 40 cortisone, hydrocortisone acetate, hydrocortisone cypi chemoattractant protein, macrophage-inflammatory onate, hydrocortisone sodium phosphate, hydrocorti protein, SIS (small inducible secreted) proteins, platelet Sone sodium Succinate, prednisone, prednisolone, factor, platelet basic protein, melanoma growth stimu prednisolone acetate, prednisolone sodium phosphate, lating activity, epidermal growth factor, transforming prednisolone tebutate, prednisolone pivalate, triamcino growth factor (alpha), fibroblast growth factor, platelet 45 lone, triamcinolone acetonide, triamcinolone hexac derived endothelial cell growth factor, insulin-like etonide, triamcinolone diacetate, methylprednisolone, growth factor, nerve growth factor, vascular endothelial methylprednisolone acetate, methylprednisolone growth factor, bone morphogenic proteins, and bone Sodium Succinate, flunsolide, beclomethasone dipropi growth/cartilage-inducing factor (alpha and beta). Other onate, betamethasone sodium phosphate, betametha cell response modifiers (Such as the interleukins, inter 50 Sone, Vetamethasone disodium phosphate, vetametha leukin inhibitors or interleukin receptors, including Sone sodium phosphate, betamethasone acetate, interleukin 1 through interleukin 10; interferons, includ betamethasone disodium phosphate, chloroprednisone ing alpha, beta and gamma; hematopoietic factors, acetate, corticosterone, desoxycorticosterone, desoxy including erythropoietin, granulocyte colony stimulat corticosterone acetate, desoxycorticosterone pivalate, ing factor, macrophage colony stimulating factor and 55 desoximethasone, estradiol, fluorocortisone, fluorocor granulocyte-macrophage colony stimulating factor; tisone acetate, dichlorisone acetate, fluorohydrocorti tumor necrosis factors, including alpha and beta; trans Sone, fluorometholone, fluprednisolone, parametha forming growth factors (beta), including beta-1, beta-2, SOne, paramethasone acetate, androsterone, beta-3, inhibin, and activin) Therapeutic enzymes (Such fluoxymesterone, aldosterone, methandrostenolone, as proteases, phospholipases, lipases, glycosidases, cho 60 methylandrostenediol, methyl testosterone, nore lesterol esterases, and nucleases) Peptide-nucleic acid thandrolone, testosterone, testosterone enanthate, test (PNA) conjugate, polysaccharide-peptide conjugates osterone propionate, equilenin, equilin, estradiol ben Such as glyosylated polypeptides; glycoproteins), a poly Zoate, estradiol dipropionate, estriol, estrone, estrone (ethyleneglycol)-polypeptide conjugate (PEG-ylated benzoate, acetoxypregnenolone, anagestone acetate, polypeptides), or polymer pharmaceuticals. 65 chlormadinone acetate, fluorogestone acetate, Antibodies and antibody fragments (Such as, but are not hydroxymethylprogesterone, hydroxymethylprogester limited to, therapeutic antibodies include trastuzumab, one acetate, hydroxyprogesterone, hydroxyprogester US 9,096,703 B2 77 78 O acetate, hydroxyprogesterone caproate, combination, i.e., a redox pair, as the polymerization initiator melengestrol acetate, normethisterone, pregnenolone, in a redox free radical polymerization. progesterone, ethynyl estradiol, mestranol, dimethister In some embodiments, it is preferable that initiators and one, ethisterone, ethynodiol diacetate, norethindrone, ligands often used in ATRP Such as bromine-containing ini norethindrone acetate, norethisterone, fluocinolone tiators and ligands such as bipyridine are not used in the acetonide, flurandrenolone, hydrocortisone sodium Suc process as they may be non-biocompatible at certain levels. In cinate, methylprednisolone sodium Succinate, predniso further embodiments, it is preferred not to have a detectable lone phosphate Sodium, triamcinolone acetonide, level of bipyridine in the polymer modified article or in aque hydroxydione sodium, Spironolactone, Oxandrolone, ous or organic extractions of the polymer modified article. In oxymetholone, prometholone, testosterone cypionate, 10 further embodiments, it is preferred not to have a detectable level of bromine in the polymer modified article or in aqueous testosteronephenylacetate, estradiol cypionate, and nor or organic extractions of the polymer modified article. Bipy ethynodrel, analogs thereof, or combinations thereof) ridine and bromine can be detected with HPLC or UV analy Non-steroidal anti-inflammatory agents including their S1S. racemic mixtures or individual enantiomers where 15 The general procedure described herein can be modified as applicable (Such as ibuprofen, flurbiprofen, ketopro necessary to accommodate different Substrate materials, ini fen, aclofenac, diclofenac, aloxiprin, aproxen, aspirin, tiators systems, and/or monomer compositions and to incor diflunisal, fenoprofen, indomethacin, mefenamic acid, porate high concentrations of the initiator into and/or onto the naproxen, phenylbutaZone, piroxicam, salicylamide, Substrate or undercoating layer. High initiator concentrations Salicylic acid, Sulindac, desoxysulindac, tenoxicam, tra may result in highly densely coated Surfaces which improves madol, ketoralac, flufenisal, Salsalate, triethanolamine the non-fouling activity of the composition. For example, Salicylate, aminopyrine, antipyrine, oxyphenbutaZone, highly densely coated Surfaces contain polymer chains that apaZone, cintaZone, flufenamic acid, clonixerl, clonixin, reduce penetration of fouling molecules into the coating. meclofenamic acid, flunixin, coichicine, demecolcine, Without being bound to any particular theory it is presently allopurinol, oxypurinol, benzydamine hydrochloride, 25 theorized that a reservoir of initiator incorporated in the sub dimefadane, indoxole, intrazole, mimbane hydrochlo strate may enhance re-initiation and branching of non-fouling ride, paranylene hydrochloride, tetrydamine, benzin polymer from the surface and near the surface of the substrate. dopyrine hydrochloride, fluprofen, ibufenac, naproXol, This re-initiation, in turn, may increase the thickness of the fenbufen, cinchophen, diflumidone sodium, fenamole, non-fouling polymer (in other words, the distance the non flutiazin, metazamide, letimide hydrochloride, nexeri 30 fouling polymer stretches above the substrate in a direction dine hydrochloride, octaZamide, molinazole, neocin normal to the substrate surface) as well as the degree of chophen, nimazole, proxazole citrate, tesicam, tesimide, branching. tolimetin, and triflumidate). In general, and as described in greater detail elsewhere Formulations of the above antimicrobial or antiseptic herein, incorporation of initiator into the substrate enables agents may be enhanced by altering the solubility or physical 35 polymeric material to be grafted from the substrate surface characteristics of the agent if salts or crystals are used, for and from within the near-surface Zone beneath the substrate instance by using nanoparticles or other formulations with surface. In general, however, it is preferred that the grafted reduced size or enhanced Surface area per mass. polymeric material not extend too far into the Substrate; thus, Non-fouling Surfaces, such as Zwitterionic Surfaces, may in one embodiment grafted polymeric material is present in also present a particularly attractive Surface for immobiliza 40 the near-Surface Zone but not at greater depths, i.e., not in the tion of biomolecules, such as antibodies, for use as biosen substrate bulk. The maximum depth to which near-surface sors. Immobilized antibodies on non-fouling Surface Sur Zone extends, i.e., the distance of the lower boundary of the faces, such as Zwitterionic Surfaces, have been demonstrated near-surface Zone as measured from the Substrate Surface is, to retain both antibody activity and antigen specificity in at least in part, a function of the initiator and the technique whole blood. “Smart” implanted medical devices that detect 45 used to incorporate initiator in the Substrate. Typically, how undesirable activation of specific immune pathways, such as ever, it is generally preferred that the lower boundary not be proinflammatory cytokines, or the presence of a possible greater than 20 micrometers from the substrate surface. By infectious agent, perhaps through detection of a secreted way of example, the lower boundary may not be greater than microbial toxin, could be designed, for example, by utilizing 15 micrometers from the substrate surface. By way of further specific antibodies or biomolecules tailored to monitor these 50 example, the lower boundary may not be greater than 10 threats. Appropriate therapeutic strategies could then be micrometers from the substrate surface. Similarly, the mini employed before an unfavorable outcome, such as infection, mum depth of near-Surface Zone, i.e., the distance of the upper arises. The stability of the Zwitterionic molecule in vivo pro boundary of the near-surface Zone from the substrate surface vides a unique advantage in this type of scenario due to its is, at least in part, also a function of the initiator and the longevity. 55 technique used to incorporate initiator in the Substrate. Typi Polymerization cally, however, the upper boundary will be at least 0.1 The polymeric surface modifications of the present inven micrometers from the substrate surface. By way of example, tion may be formed by synthetic means including, but not the upper boundary may be at least 0.2 micrometers from the limited to, free radical polymerization, ionic polymerization, substrate surface. By way of further example, the upper atom transfer radical polymerization (ATRP), nitroxide 60 boundary may be at least 0.3 micrometers from the substrate mediated polymerization (NMP), reversible addition-frag Surface. mentation polymerization (RAFT), ring opening metathesis The quality of the surface modification formed in the poly polymerization (ROMP), telluride mediated polymerization merization process is, at least in part, influenced by the quality (TERP) or acyclic diene metathesis polymerization (AD of the surface of the substrate prior to polymerization. For MET), and UV, thermal, or redox free radical initiated poly 65 example, prior to polymerization, the Surface may be con merization. In a preferred embodiment, the polymer is taminated, intentionally or otherwise, with particles, waxes formed using an oxidizing agent and a reducing agent, in and other compositions that may remain on the Surface of the US 9,096,703 B2 79 80 Substrate as an artifact of the manufacturing process, Subse cocamidopropyl betaine; lecithin. Some preferred Surfactants quent handling of the Substrate, and/or as part of the intended also include nonionic Surfactants such as fatty alcohols: cetyl Substrate composition. The Substrate Surface may also alcohol, Stearyl alcohol, cetostearyl alcohol (consisting pre include significant Surface roughness, physical defects Such dominantly of cetyl and Stearyl alcohols), oleyl alcohol; poly as Scratches, pinholes, or Voids, and chemical defects, such as oxyethylene glycol alkyl ethers (Bri): CH-(CH2). particle(s) of radiopacifing agents (such as barium Sulfate, (OCH) OH: octaethylene glycol monododecyl ether, bismuth oxychloride, bismuth subcarbonate, bismuth triox pentaethylene glycol monododecyl ether, Polyoxypropylene ide, lanthanum oxide, tantalum pentoxide, and metallic gold, glycol alkyl ethers: CH(CH2)(OCH) OH: Gluco silver, platinum, palladium, tungsten, and tantalum) that are side alkyl ethers: CH (CH) (O-Glucoside)-H; Decyl only partially contained within the substrate. For example, 10 glucoside, Lauryl glucoside, Octyl glucoside; Polyoxyethyl Substrates containing barium Sulfate typically have some ene glycoloctylphenol ethers: CH(CH4)(OCH) as OH: barium sulfate particles that are partially contained within the Triton X-100; Polyoxyethylene glycol alkylphenol ethers: Substrate and partially exposed; the exposed portions of Such CH3(CH4)(OCH) OH: Nonoxynol-9, Glycerol alkyl barium sulfate particles may extend from the surface of a esters: Glyceryl laurate; Polyoxyethylene glycol sorbitan Substrate to a height of as much as 1 micrometer (as measured 15 alkyl esters: Polysorbates: Sorbitan alkyl esters: Spans; Coca from the surface of the substrate using SEM). mide MEA, cocamide DEA. Dodecyldimethylamine oxide: In accordance with one embodiment, the Substrate Surface Block copolymers of polyethylene glycol and polypropylene is preferably pre-treated prior to polymerization. For glycol: Poloxamers. example, the Substrate surface may be cleaned using water, Alternatively, or additionally, the substrate may be chemi Solvents, Surfactants, enzymes, or other cleaning solutions or cally, mechanically or chemomechanically polished prior to gases to remove particles, waxes or other foreign composi polymerization to reduce Surface roughness, reduce the inci tions that may be on or near the surface of the substrate. dence and/or severity of cracks, pinholes and other structural Alternatively, or additionally, the substrate surface may be defects in the substrate surface. For example, the substrate mechanically, chemically or chemomechanically treated to may be solvent polished by exposing the Substrate to a vapor reduce the incidence and/or the severity of physical and 25 of a solvent such as chloroform, dioxane or tetrahydrofuran. chemical defects. After polishing the substrate surface preferably has a global In one embodiment, the substrate is treated prior to poly average R. Surface roughness that is less than the global merization with a composition Such as an acid, base, chelator average R. Surface roughness of the unpolished substrate. or reactant that dissolves or chemically reacts with and By way of further example, in one embodiment the polished reduces the concentration of any compositions that are 30 Substrate surface has a global average R. Surface roughness included as chemical defects, or even swells the substrate that is no more than 90% of the global average R. Surface allowing the particles to be released from the substrate. For roughness of the unpolished substrate surface. By way of example, exposed portions of barium Sulfate particles may be further example, in one embodiment the polished substrate partially or completely dissolved using a mineral or organic Surface has a global average R. Surface roughness that is no acid and optionally, a chelator. In one Such exemplary 35 more than 75% of the global average R. Surface roughness embodiment, polyurethane comprising particles of barium of the unpolished substrate surface. By way of further sulfate may be treated with hydrochloric acid to at least par example, in one embodiment the polished substrate Surface tially remove exposed barium Sulfate particles. has a global average R. Surface roughness that is no more In one embodiment, the substrate is treated prior to poly than 50% of the global average R. Surface roughness of the merization with a Surfactant to remove particles, waxes or 40 unpolished Substrate Surface. other foreign compositions that may be on or near the Surface Alternatively, or additionally, in one embodiment the sub of the substrate. Some preferred surfactants include anionic strate is precoated prior to polymerization with any of the Surfactants, such as alkyl Sulfates: ammonium lauryl Sulfate, compositions identified herein as a precoating or undercoat sodium lauryl sulfate (SDS, sodium dodecyl sulfate, another ing compositions to cover physical defects and/or reduce the name for the compound); alkyl ether Sulfates: Sodium laureth 45 Surface roughness of the Substrate surface. In general, the sulfate, also known as sodium lauryl ether sulfate (SLES), precoat preferably has an average thickness that equals or Sodium myreth Sulfate; Sulfonates: for example docusates: exceeds the global average R. Surface roughness of the dioctyl sodium SulfoSuccinate; Sulfonate fluoroSurfactants: uncoated Substrate. For example, in one embodiment, the perfluorooctanesulfonate (PFOS), perfluorobutanesulfonate; precoat has an average thickness that is at least 110% of the alkylbenzene Sulfonates; phosphates: for example alkyl aryl 50 global average R. Surface roughness of the uncoated Sub ether phosphate, alkyl ether phosphate; carboxylates: for strate. By way of further example, in one embodiment, the example alkyl carboxylates: fatty acid salts (soaps): sodium precoat has an average thickness that is at least 200% of the Stearate; sodium lauroyl sarcosinate; carboxylate fluorosur global average R. Surface roughness of the uncoated Sub factants: perfluorononanoate, perfluorooctanoate (PFOA or strate. By way of further example, in one embodiment, the PFO). Some preferred surfactants also include cationic sur 55 precoat has an average thickness that is at least 300% of the factants, such as octenidine dihydrochloride; alkyltrimethy global average R. Surface roughness of the uncoated Sub lammonium salts: cetyl trimethylammonium bromide strate. By way of further example, in one embodiment, the (CTAB) a.k.a. hexadecyl trimethyl ammonium bromide, precoat has an average thickness that is at least 400% of the cetyltrimethylammonium chloride (CTAC); cetylpyridinium global average R. Surface roughness of the uncoated Sub chloride (CPC); polyethoxylated tallow amine (POEA); ben 60 strate. In addition, the precoating preferably reduces the glo Zalkonium chloride (BAC); benzethonium chloride (BZT): bal average R. Surface roughness of the substrate surface. 5-bromo-5-nitro-1,3-dioxane; dimethyldioctadecylammo Stated differently, the precoated substrate surface preferably nium chloride; dioctadecyldimethylammonium bromide has an average thickness that equals or exceeds the global (DODAB). Some preferred surfactants also include Zwitteri average R. Surface roughness of the uncoated Substrate and onic (amphoteric) surfactants: such as CHAPS (3-(3-Chola 65 a global average R. Surface roughness that is less than the midopropyl)dimethylammonio-1-propanesulfonate); coca global average R. Surface roughness of the Substrate prior to midopropyl hydroxysultaine; amino acids; Imino acids; the application of the precoat. For example, in one embodi US 9,096,703 B2 81 82 ment the precoated Substrate Surface has an average thickness dipping the external portion in a polymer Solution or disper that is at least 110% of the global average R. Surface rough sion to coat the external portion and flowing a polymer Solu ness of the uncoated Substrate and a global average R. tion or dispersion through the intralumenal portion to coat the surface roughness that is no more than 90% of the global intralumenal portion. Coating application parameters utilized average R. Surface roughness of the Substrate prior to the 5 to effect coating control include the solvent system, percent application of the precoat. By way of further example, in one Solids and viscosity, and cure temperature and time. Suitable embodiment the precoated Substrate Surface has an average solvents for the polymer primer layer include, but are not thickness that is at least 110% of the global average R. limited to, alcohols, such as methanol or ethanol. Application Surface roughness of the uncoated Substrate and a global and cure temperature can vary, for example between ambient average R. Surface roughness that is no more than 75% of 10 the global average R. Surface roughness of the Substrate and 50° C. so as not to affect physical properties of the prior to the application of the precoat. By way of further underlying Substrate, for example, a polyurethane Substrate. Solids content can vary between 0.5-10%, with solution vis example, in one embodiment the precoated Substrate Surface cosity no higher than 12 cB for ease of handling and applica has an average thickness that is at least 110% of the global tion. average R. Surface roughness of the uncoated Substrate and 15 a global average R. Surface roughness that is no more than The average thickness of a polymeric Surface modification 50% of the global average R. Surface roughness of the or coating on a Substrate can be approximated using attenu Substrate prior to the application of the precoat. ated total reflectance (ATR) infrared spectrometry if the infra Regardless of the pre-treatment steps, or even whether red spectra and refractive indices of the typical polymeric pre-treatment steps are employed, the Surface of the Substrate Surface material and the typical Substrate material can be from which the non-fouling material is to be grafted has a determined independently and if the range of the modification global average R. Surface roughness that is no more than or coating thickness is between 10 nm and 5000 nm. A matrix 100 nm. In certain embodiments, the surface is even of synthetic infrared absorbance spectra can be constructed Smoother. For example, the Surface may have a global average using the principal component spectra (those of the coating R. Surface roughness of less than 50 nm. In some embodi 25 material and the substrate material) and Beer's law (A=ebC) ments, the Surface may have a global average R. Surface where b, the optical pathlength, is replaced by the exponen roughness of less than 20 nm. tially decaying and wavelength dependent depth of penetra Additionally, or alternatively, and regardless of the pre tion of the ATR evanescent wave. An empirically measured treatment steps, or even whether pre-treatment steps are sample is then compared across all the synthetic spectra in the employed, the surface of the substrate from which the non 30 matrix and the closest match, determined by the minimum fouling material is to be grafted has a visually observable n-dimensional cosine statistical distance, is the one of the surface defect density (i.e., visually observable number over sample's polymeric surface modification or coating thick a field size of 20x20 micrometers) of defects having a size CSS. (i.e., alongest dimension) greater than about 0.5 micrometers In one embodiment, for example, the average thickness of that is less than 0.1 defects/um. For example, the surface of 35 a homopolymeric SBMA (N-(3-sulfpropyl)-n-methacry the substrate from which the non-fouling material is to be loxyethyl-n, n-dimethylammonium betaine) hydrogel Surface grafted may have a surface defect density of defects having a modification or coating on a polyetherurethane plus 10% to size greater than about 0.5 micrometers that is less than 0.05 50% BaSO substrate can be determined using attenuated defects/um. By way of further example, the surface of the total reflectance (ATR) infrared spectrometry if the range of substrate from which the non-fouling material is to be grafted 40 the modification or coating thickness is between 10 nm and may have a Surface defect density of defects having a size 5000 nm and the BaSO content of the substrate is constant to greater than about 0.5 micrometers that is less than 0.01 within +/-5%. The value of the absorbance of the vibrational defects/um. By way of further example, the surface of the SO3 stretch at 1037.0 cm (point baseline corrected by sub substrate from which the non-fouling material is to be grafted tracting the absorbance value at 994.7 cm) divided by the may have a Surface defect density of defects having a size 45 value of the absorbance of the urethane peak at 1309.5 cm greater than about 0.5 micrometers that is less than 0.002 (point baseline corrected by subtracting the absorbance value defects/um. By way of further example, the surface of the at 1340.0 cm) equals a value relative to the concentration of substrate from which the non-fouling material is to be grafted SBMA present. By taking the natural log of the relative value, may have a Surface defect density of defects having a size adding 0.1641 and then multiplying by 500 yields a value that greater than about 0.5 micrometers that is less than 0.001 50 correlates to the thickness of the homopolymeric hydrogel defects/um. Surface modification or coating as determined by the Syn In one presently preferred embodiment, the substrate is thetic ATRIR matrix described above. precoated with any of the precoating or undercoating mate By way of further example, the average thickness of a rials described elsewhere herein. In one such embodiment, homopolymeric SBMA (N-(3-sulfpropyl)-n-methacryloxy the precoat typically has an average thickness of at least about 55 ethyl-n, n-dimethylammonium betaine) hydrogel Surface 100 nm. In some embodiments, the precoat will be substan modification or coating on a polyetherurethane Substrate can tially thicker, for example, the precoat may have an average be determined using attenuated total reflectance (ATR) infra thickness of as much as 500 micrometers. In general, how red spectrometry if the range of the modification or coating ever, the precoat will be thinner. For example, the precoat may thickness is between 10 nm and 5000 nm. The value of the have an average thickness of about 1-50 micrometers. By way 60 absorbance of the vibrational SO3 stretch at 1037.0 cm of further example, the precoat may have an average thickness (point baseline corrected by subtracting the absorbance value of about 10-30 micrometers. at 994.7 cm) divided by the value of the absorbance of the In some instances, the Substrate will have a complex shape urethane peak at 1309.5 cm (point baseline corrected by or geometry with inner and outer Surfaces to be coated. For subtracting the absorbance value at 1340.0 cm) equals a example, multi-lumen catheters have an exterior Surface and 65 value relative to the concentration of SBMA present. By two or more longitudinal lumens that may be coated. Poly taking the natural log of the relative value, adding 0.9899 and meric primer coatings may be applied by simultaneously then multiplying by 500 yields a value that correlates to the US 9,096,703 B2 83 84 thickness of the homopolymeric hydrogel Surface modifica tain embodiments, the substrate polymer from which the tion or coating as determined by the synthetic ATRIR matrix non-fouling polymer will be grafted will not swell more than described above. 15% by volume at 25°C. under equilibrium conditions in the In a preferred embodiment, some consideration is given to polymerization mixture solvent system. In certain embodi the combined thickness of the undercoating and the grafted 5 ments, the Substrate polymer from which the non-fouling polymer layer. For example, it is generally preferred that the polymer will be grafted will not swell more than 5% by undercoating and the grafted polymer not materially change volume at 25°C. under equilibrium conditions in the poly the dimensions of the components of a devices, such as lumen merization mixture solvent system. In certain embodiments, diameters. Thus, in Some embodiments, the combined global the substrate polymer from which the non-fouling polymer average dry thickness of the undercoating and the grafted 10 will be grafted will not swell or may even shrink at 25°C. polymer layer is <1% of the diameter of a catheter lumen in under equilibrium conditions in the polymerization mixture which it is applied. In some embodiments, the global average Solvent system. As previously noted, the Substrate may be a dry thickness of the undercoating and the grafted polymer composite of materials. In Such instances, it is preferred that layer is <0.5% of the diameter of a catheter lumen in which it the near-surface region of the substrate into which the poly is applied. In some embodiments, the global average dry 15 merization initiator is incorporated satisfy the Swelling crite thickness of the undercoating and the grafted polymer layer is ria recited herein. For example, in those embodiments in <0.25% of the diameter of a catheter lumen in which it is which the Substrate comprises a coating of a precoat material applied. In further embodiments, the global average dry overlying a metal, ceramic, glass or semi-metallic material, it thickness of the undercoating and the grafted polymer layer is is preferred that the coating of the precoat material not swell <0.1% of the diameter of a catheter lumen in which it is 20 more than 30% by volume at 25°C. under equilibrium con applied. In certain embodiments, the global average dry ditions in the polymerization mixture solvent system. thickness of the undercoating and the grafted polymer layer is The initiator(s) incorporated into the substrate preferably <0.05% of the diameter of a catheter lumen in which it is have limited solubility in the solvent system comprised by the applied. In further embodiments, the global average dry polymerization mixture and include any of the initiators iden thickness of the undercoating and the grafted polymer layer is 25 tified herein. In general, it is preferred that the incorporated <0.01% of the diameter of a catheter lumen in which it is initiator(s) have a 10 hour T/2 decomposition temperature of applied. In further embodiments, the global average dry 25-175° C. In one particular embodiment, the incorporated thickness of the undercoating and the grafted polymer layer is initiator(s) have a 10 hour T/2 decomposition temperature of <0.001% of the diameter of a catheter lumen in which it is 70-130°C. Advantageously, having a 10 hour T/2 decompo applied. 30 sition temperature of 70-130° C. tends to increase the density To induce Small polymerization initiator molecules to con of interfacial initiation events from the redox reaction and centrate at or near the substrate surface, where polymeriza effectively outcompete thermal initiation. tion is initiated and propagated, polymerization mixture sol As described elsewhere herein, the initiator may comprise vent systems with Surface tensions of a magnitude differing a redox pair; in Such embodiments, at least one member of from the surface energy of the substrate and one or more 35 such pair have such a limited solubility in the polymerization polymerization initiators having limited solubility in the mixture solvent system. In one embodiment, both members polymerization mixture solvent system are selected. The Sur of the redox pair have limited solubility in the polymerization faces of the substrate from which the non-fouling material is mixture solvent system. In an alternative embodiment, one to be grafted surfaces may be hydrophobic or hydrophilic, member of the pair is soluble in the polymerization mixture and the polymerization mixture solvent system may be acque- 40 solvent system but the other has limited solubility in the ous, comprise polar organic solvents, aqueous mixtures of polymerization mixture solvent system. Without being bound polar organic Solvents, or aqueous mixtures of any organic to any particular theory, it is presently believed that when one compound designed to modify the Surface tension of aqueous member of a redox pair is soluble in the polymerization solutions. Optionally, for hydrophobic substrates, hydropho mixture solvent system and the other has limited solubility in bic initiator(s) and hydrophilic solvent systems, e.g., aqueous 45 the polymerization mixture solvent system, the two are phase media are selected. Preferably, if the substrate is hydrophilic, separated and initiation is enhanced at the interface of the two at least one hydrophilic initiator and a non-polar organic phases which tends to decrease solution polymerization and Solvent system is selected. increase grafting at or near the Substrate surface. Thus, for Preferably, the substrate (or at least the portion of the example, either member of the redox pair may be hydropho Substrate into which the polymerization initiator is incorpo- 50 bic and either member of the pair may be hydrophilic, pro rated) is not significantly Swelled by the polymerization mix vided at least one of the members has limited solubility in the ture (e.g., by the polymerization mixture solvent system, the polymerization mixture solvent system. In one preferred polymerization monomers, or both) and the initiator(s) incor embodiment, a hydrophobic oxidizer is paired with a hydro porated into the substrate has/have limited solubility in the philic reducing agent. In another preferred embodiment, a solvent system. As a result, the interface between substrate 55 hydrophilic oxidizer is paired with a hydrophobic reducing Surface and the polymerization mixture can have a relatively agent. For example, in one embodiment, the redox pair com high local concentration of initiator(s) to initiate non-fouling prises a peroxide and a reducing agent wherein the peroxide polymer growth from or near the Substrate Surface and to has limited solubility in the polymerization solvent system (re)initiate polymer growth from the grafted non-fouling and the reducing agent has high solubility in the polymeriza polymer. Without being bound to any particular theory, it is 60 tion solvent system. By way of further example, in certain presently believed that this approach leads to the grafting of a embodiments, the peroxide has a log P partition coefficient relatively highly branched non-fouling polymer from the sub greater than or equal to 3 for hydrophobic Substrates and Strate. phases and a log P partition coefficient less than 3 for hydro In a preferred embodiment, the substrate polymer from philic Substrates and phases. By way of further example, in which the non-fouling polymer will be grafted will not swell 65 certain embodiments, the peroxide has a log P partition coef more than 30% by volume at 25°C. under equilibrium con ficient greater than or equal to 5 for hydrophobic substrates ditions in the polymerization mixture solvent system. In cer and phases and a log P partition coefficient less than 1 for US 9,096,703 B2 85 86 hydrophilic substrates and phases. By way of further previous initiator imbibing period. In general, the Substrate is example, in certain embodiments, the peroxide has a log P immersed in the initiator-containing Solution for at least sev partition coefficient greater than or equal to 7 for hydrophobic eral seconds before polymerization is initiated. In some Substrates and phases and a log P partition coefficient less embodiments, the Substrate is immersed in the initiator-con than -1 for hydrophilic substrates and phases. By way of 5 taining Solution for longer times. For example, the Substrate further example, in certain embodiments, the peroxide has a may be immersed in the initiator-containing Solution for at log Ppartition coefficient greater than or equal to 9 for hydro least several minutes. By way of further example, the sub phobic Substrates and phases and a log P partition coefficient strate may be immersed in the initiator-containing Solution less than -3 for hydrophilic substrates and phases. for at least about 15 minutes before polymerization is initi In one embodiment, an initiator is incorporated into the 10 ated. In some embodiments, the substrate will be immersed in Substrate by initially incorporating an initiator-precursor into the initiator-containing Solution for at least 1 hour at room the Substrate and activating the initiator-precursor to an ini temperature or elevated temperatures for initiators having a tiator. 10 hour T/2 decomposition temperature of 70-130° C. before In accordance with one aspect of the present invention, the polymerization is initiated. In further embodiments, the sub polymerization initiator(s) may be incorporated into and/or 15 strate will be immersed in the initiator-containing Solution for onto the Substrate by various techniques. In one such method, at least 2 hour before polymerization is initiated. In yet further the Substrate (including Substrates having precoat or under embodiments, the substrate will be immersed in the initiator coat as previously described) is imbibed with the polymer containing solution for at least 16 hour before polymerization ization initiator; that is, the polymerization initiator is is initiated. Depending upon the time, temperature and con absorbed into the substrate. In one embodiment, the initia centration of initiator in the initiator-containing Solution, a tor(s), i.e., an initiator or a mixture of different initiators, is concentration gradient of initiator in the Substrate may be introduced into and/or onto the substrate's surface by physio established. In some embodiments, it may be preferable to adsorption, wherein the initiator is dissolved in a solvent or have a higher concentration of initiator in the Substrate nearer combination of solvents and the substrate (with or without an to the Surface. As noted, the initiator may be presentina range undercoating layer) is Submerged in the mixture for a time 25 of concentrations in the initiator-containing Solution. In gen and at a temperature to achieve sufficient absorption by the eral, the concentration of the initiator in the initiator-contain substrate. The substrate is allowed to swell ultimately imbib ing solution will be at least 0.01% by weight. For example, in ing initiator into the Substrate. In general, the amount of some embodiments, the concentration of the initiator will initiator incorporated into a substrate during the Soak will, at generally be at least 0.1% by weight. In some embodiments, least in part, be a function of the solubility of the initiator in 30 the concentration will be even greater, e.g., at least 0.5% by the solvent system, solubility of the initiator in the substrate weight. In some embodiments, the concentration will be even as well as the soak time, temperature and concentration of the greater, e.g., at least 1% by weight. In some embodiments, the initiator in the Solution, as well as the chemical composition concentration will be even greater, e.g., at least 10% by of the substrate and the initiator. weight. In certain exemplary embodiments, the concentration In a preferred embodiment, the surface of the substrate to 35 of the initiator in the initiator-containing solution will be in be imbibed with the polymerization initiator(s) comprises a the range of about 0.2 to about 1% by weight. In certain polymer, natural or synthetic. In an alternative embodiment, exemplary embodiments, the concentration of the initiator in the substrate is an imbibable material selected from among the initiator-containing solution will be in the range of about polymers, natural or synthetic, biological tissues, living or 0.2 to about 10% by weight. In certain exemplary embodi dead, woven non-woven fibers, and combinations thereof. 40 ments, the concentration of the initiator in the initiator-con Certain (uncoated) Substrates such as a metal, ceramic, glass, taining solution will be in the range of about 0.5 to about 5% and semi-metallic Substrates lack the capacity to absorb Suf by weight. In certain exemplary embodiments, the concen ficient initiator. In general, therefore, for these substrates it is tration of the initiator in the initiator-containing Solution will preferred to precoat the Surface of the metal, ceramic, glass or be in the range of about 0.75 to about 3% by weight. In each semi-metallic Substrate with an undercoating or precoating, 45 of these embodiments, the initiator is preferably one of the from which the polymeric material may be grafted. For UV, thermal or redox initiators described elsewhere herein. example, metal, ceramic, glass, and semi-metallic Substrates As a result of the imbibing process, the imbibed substrate may be precoated with a polymer selected from polyamide, may contain about 0.001% by weight initiator. In some polyamine, polyanhydride, polyazine, poly(carbonate), poly embodiments, the imbibed substrate will contain greater ester, polyether, polyetheretherketone (PEEK), polyguani 50 amounts of initiator, e.g., at least about 0.01% by weight. For dine, polyimide, polyketal, poly(ketone), polyolefin, poly example, in some embodiments the imbibed substrate will (orthoester), polyphosphazine, polysaccharide, polysiloxane, contain at least about 0.1% by weight. By way of further polysulfone, polyurea, polyurethane, halogenated polymer, example, in some embodiments the imbibed substrate will silicone, aldehyde crosslinked resin, epoxy resin, phenolic contain about 0.05% to about 2% by weight initiator. By way resin, latex, or a copolymer or blend thereof, and the pre 55 of further example, in some embodiments the imbibed sub coated substrate is then imbibed as previously described. strate will contain about 0.1% to about 1% by weight initiator. The quantity of initiator introduced to the substrate can be By way of further example, in some embodiments the controlled by changing the concentration of the initiator in the imbibed substrate will contain about 0.2% to about 0.5% by Solvent Solution and/or by changing the amount of time the weight initiator. By way of further example, in some embodi Substrate is allowed to Soak in the initiator Solution during one 60 ments the imbibed substrate will contain about 1% to about initiator imbibing period or by repeating any number of ini 10% by weight initiator. Typically, however, the imbibed tiator imbibing periods as required. Temperature is not nar substrate will contain less than about 20% by weight initiator. rowly critical, with temperatures in the range of room tem In each of these embodiments, the initiator is preferably one perature to elevated temperatures being typical. When of the UV, thermal or redox initiators described elsewhere utilizing multiple periods of initiator imbibing, the initiator 65 herein. The solvent used to imbibe the substrate with initiator used in the Subsequent imbibing periods can be the same as, may have the capacity to swell the substrate (or at least the different from, or a mixture with the initiator used in the portion of the substrate to be imbibed with initiator) to various US 9,096,703 B2 87 88 degrees. Typically, the imbibing solvent has a capacity to will have a thickness of about 100 nm to about 500 microme swell the substrate (or at least the portion of the substrate to be ters. In each of these embodiments, the initiator is preferably imbibed with initiator) less than 900% by volume at room one of the UV, thermal or redox initiators described elsewhere temperature and ambient pressure. For example, in one Such herein. embodiment, the imbibing solvent has a capacity to Swell the In certain preferred embodiments, the co-deposited layer substrate (or at least the portion of the substrate to be imbibed will contain, as the co-deposited polymer, polyurethane, with initiator) less than 750% by volume. By way of further polystyrene, polyester, Sol-gels, or a combination thereof. example, in one such embodiment, the imbibing solvent has a Thus, for example, in one embodiment, the co-deposited capacity to swell the substrate (or at least the portion of the layer will have a thickness of about 100 nm to about 50 substrate to be imbibed with initiator) less than 500% by 10 micrometers, and the weight ratio of initiator to polymer in volume. By way of further example, in one such embodiment, the co-deposited layer will be at least 1:1000, respectively. In the imbibing solvent has a capacity to swell the substrate (or certain more specific embodiments, the co-deposited layer at least the portion of the substrate to be imbibed with initia will contain polyurethane as the co-deposited polymer, will tor) less than 250% by volume. By way of further example, in have a thickness of about 1-50 micrometers. By way of fur one such embodiment, the imbibing solvent has a capacity to 15 ther example, the co-deposited layer may have an average swell the substrate (or at least the portion of the substrate to be thickness of about 10-30 micrometers. By way of further imbibed with initiator) less than 100% by volume. By way of example, in each of these exemplary embodiments the co further example, in one such embodiment, the imbibing Sol deposited layer may have a weight ratio of initiator to poly vent has a capacity to swell the substrate (or at least the mer of about 1:1,000 to about 20:1, respectively. In addition, portion of the substrate to be imbibed with initiator) less than in each of these exemplary embodiments, the initiator is pref 100% by volume. By way of further example, in one such erably one of the UV, thermal or redox initiators described embodiment, the imbibing solvent has a capacity to Swell the elsewhere herein. substrate (or at least the portion of the substrate to be imbibed The solvent and/or solvent mixtures used to co-deposit the with initiator) less than 25% by volume. initiator(s) and the polymeras a precoat may have the capac In a preferred embodiment, the imbibed substrate is pref 25 ity to swell the substrate to various degrees. Typically, the erably washed using a solvent, optionally with a solvent that co-deposition solvent swells the substrate (or at least the swells that substrate, and optionally dried. In other embodi portion of the substrate to be imbibed with initiator) less than ments, the substrate is washed with solvents, which may be 900% by volume at room temperature and ambient pressure. the same or different from the imbibing solvents, or the sub For example, in one such embodiment, the co-deposition strate may not be washed. For example, the wash solvent may 30 solvent swells the substrate (or at least the portion of the swell the substrate, shrink the substrate, or neither. In one substrate to be imbibed with initiator) less than 100% by embodiment, the substrate is dried, partially dried or not volume. By way of further example, in one such embodiment, dried. Optionally, there may be a solvent exchange. the co-deposition solvent swells the substrate (or at least the In an alternative method, the initiator(s) is/are incorporated portion of the substrate to be imbibed with initiator) less than into the substrate by co-deposition of the initiator(s) as a 35 100% by volume. By way of further example, in one such component of a coating, i.e., a precoating or undercoating as embodiment, the co-deposition solvent swells the substrate described herein, on the surface of the substrate. For example, (or at least the portion of the substrate to be imbibed with a thin film of polymer and initiator are deposited onto the initiator) less than 25% by volume. In a preferred embodi Substrate by dipping the Substrate in a solution of initiator(s) ment, the co-deposited layer is preferably washed using a and polymer. Alternatively, a precoat layer of a flowable 40 solvent and/or solvent mixture, optionally with a solvent that mixture of the initiator(s) and a second material Such as a swells that substrate, and optionally dried. Alternatively, the polymeric material are deposited onto the surface of the sub co-deposited layer is preferably washed using a solvent and/ strate. The precoating may thus be applied to any of the or solvent mixture, optionally with a solvent and/or solvent Substrates described herein, including metals, ceramics, mixture that has limited Swelling of the Substrate, and option glasses, polymers, biological tissues, living or dead, woven 45 ally dried. Alternatively, the co-deposited layer is not washed and non-woven fibers, semi-metals such as silicon. For using a solvent and optionally dried. example, the metal, ceramic, glass, polymer, biological tis In one exemplary embodiment, a solution containing 1% to Sue, fiber, or semi-metal may be precoated with a polymerand 5% (wt/wt) urethane can be prepared by dissolving the appro initiator mixture wherein the polymer is selected from polya priate weight of urethane pellets in a Suitable organic solvent, mide, polyamine, polyanhydride, polyazine, poly(carbon 50 Such as tetrahydrofuran, and diluting the Solution with a sec ate), polyester, polyether, polyetheretherketone (PEEK), ond solvent, Such as methanol. The final methanol concentra polyguanidine, polyimide, polyketal, poly(ketone), polyole tion is preferably between 10%-90%, more preferably fin, poly(orthoester), polyphosphazine, polysaccharide, pol between 15%-85%, most preferably 60%. One or more suit ysiloxane, polysulfone, polyurea, polyurethane, halogenated able initiator molecules, such as benzoyl peroxide or dicumyl polymer, silicone, aldehyde crosslinked resin, epoxy resin, 55 peroxide, are added to the polymer Solutionata concentration phenolic resin, latex, or a copolymer or blend thereof. typically from about 0.25% to about 10%. However, concen In one embodiment, the amount of initiator co-deposited trations below 0.25% and above 10% can be used. Any with the polymer is relatively great. In certain embodiments, desired substrate can be exposed to the polymer/initiator for example, the weight ratio of initiator to polymer co-de Solution once or multiple times until a desired coating thick posited will be at least 1:1000, respectively. In some embodi 60 ness and/or initiator Surface concentration has been achieved. ments, the weight ratio of initiator to polymer co-deposited The solvent is typically removed, for example by evaporation, will be even greater, e.g., at least 1:100, 1:10, 1:1, 10:1, 100:1, from the coated substrate between each exposure to the solu or 1000:1 respectively. Typically, the ratio of initiator to poly tion, in a case where the Substrate is exposed multiple times. mer will be in the range of about 1:1 to about 20:1. In addition, After the final exposure, the substrate is optionally allowed to the co-deposited layers (i.e., the layers containing co-depos 65 sit for at least 10 minutes to allow any residual solvent to ited initiator and polymer) will have a thickness of at least 100 evaporate, prior to placing in a polymerization reaction mix nm. For example, in one embodiment, the co-deposited layer ture. US 9,096,703 B2 89 90 In another alternative method, the initiator(s) is/are incor example, for polymeric substrates approximately 10 mmx50 porated into and/or onto the Substrate by means of a aerosol mm or 5 mmx25 mm are appropriate. deposition or spray coating process. The initiator(s) is/are Examples of the instrumentation used for analysis includes mixed with a monosolvent, co-solvent, or mixed solvent sys high-performance liquid chromatography-photo-diode array tem and applied to the Substrate surface by means of a detection-mass spectrometry (HPLC-PDA-MS) for organics directed, charged or non-charged aerosol deposition method. analysis; gas chromatography-mass spectrometry (GC-MS) For example, the initiator(s) is/are mixed with organic solvent for organics analysis; inductively coupled plasma-optical mixture and deposited onto the Substrate surface as anaerosol emission spectroscopy or mass spectrometry (ICP-OES or by means of a compressed airspray. The amount of initiator ICP-MS) for metals analysis; and ion chromatography (IC) 10 for inorganics and ion analysis. More advanced MS detectors physio-adsorbed into and/or onto the surface of the substrate such as time-of-flight (TOF) can also be used to obtain accu can be controlled by varying the amount of time the aerosol rate mass information. Hexane and alcohol extractions are stays on the surface of substrate before the solvent evaporates analyzed, for example by GC-MS and HPLC. Water and and thus affecting the amount of initiator absorbed into the alcohol extractions are analyzed, for example by HPLC. bulk of the substrate (e.g., the longer the dwell time on the 15 The initiator or its degradation products may be quantified surface the more initiator can move into the substrate bulk and and/or detected in the substrate or grafted polymer by the visa versa). The dwell time of the aerosol on the substrate can previously described methods. These include FTIR-ATR, be controlled by varying the boiling point of the aerosol electron spectroscopy for chemical analysis (ESCA, also which is done by varying the proportion of low and high called X-ray photoelectron spectroscopy, XPS), Secondary boiling point solvents in the solvent system. Additionally, the Ion Mass Spectrometry (SIMS), and surface-enhanced amount of initiator applied onto and/or into the Substrate can Raman spectroscopy (SERS). For example, peroxide may be be controlled by varying the aerosol flow rate, aerosol gas detected spectrophotometically using any of the following mixture, aerosol droplet size, aerosol charge, Substrate three methods: the iodide method (oxidation of sodium iodide charge, aerosol deposition environment (temperature, pres by peroxides in the presence of ferric chloride), the DPPH Sure, and/or atmosphere), and the amount of aerosol applied. 25 method (treatment with 1,1-diphenyl-2-picrylhydrazyl, a The aerosol deposition may be applied to any of the substrates radical scavenger, to decompose the peroxides), or the per described herein, including metals, ceramics, glasses, poly oxidase method (reduction with glutathione, catalyzed by mers, biological tissues, living or dead, woven and non-wo glutathione peroxidase, followed by measuring the coupled ven fibers, semi-metals such as silicon. oxidation of NADPH in the presence of glutathione reduc Regardless of the method of incorporation, initiator is 30 tase). See, for example, Fujimoto et al., Journal of Polymer incorporated into the substrate by imbibing the substrate or Science Part A: Polymer Chemistry, Vol. 31, 1035-1043 depositing a coating containing the initiator onto the sub (1993). strate. The incorporated initiator may comprise one initiator Similarly, the initiator(s) and/or the degradation products species, or more than one initiator species. For example, one thereof may also be extracted from the substrate/grafted poly or more species of ultraviolet (UV) initiators, one or more 35 merusing a Suitable solvent such as water, acetone or ethanol species of thermal initiators, and/or one or more species of and quantified and/or detected in the Substrate or grafted redox initiators may be incorporated into the substrate. More polymer by the previously described methods. These include specifically, in one presently preferred embodiment, the ini FTIR-ATR, electron spectroscopy for chemical analysis tiator(s) are/is incorporated into the near-Surface Zone (ESCA, also called X-ray photoelectron spectroscopy, XPS), between its upper and lower boundaries as described else 40 Secondary Ion Mass Spectrometry (SIMS), and surface-en where herein. Based upon experimental evidence to date, and hanced Raman spectroscopy (SERS). For example, peroxide without being bound to any particular theory, it appears that may be detected spectrophotometically using any of the fol the incorporated initiator permits a grafting of the polymeric lowing three methods: the iodide method (oxidation of material from within the near-surface Zone as well as the sodium iodide by peroxides in the presence of ferric chlo Substrate Surface. 45 ride), the DPPH method (treatment with 1,1-diphenyl-2-pic Regardless of the theory, it is generally preferred that the rylhydrazyl, a radical scavenger, to decompose the peroX amount of initiator incorporated into the substrate be suffi ides), or the peroxidase method (reduction with glutathione, cient to enable its detection in the substrate, prior to polymer catalyzed by glutathione peroxidase, followed by measuring ization, and detection of it or a degradation product thereof in the coupled oxidation of NADPH in the presence of glu the Substrate post-polymerization. In general, extractions can 50 tathione reductase). See, for example, Fujimoto et al., Journal use both nonpolar and polar solvents. For example, extraction of Polymer Science Part A: Polymer Chemistry, Vol. 31, Solvents such as water, acetone or ethanol; and/or other 1035-1043 (1993). extraction solvents in which the solubility of the initiator In another embodiment, quantification and/or detection of and/or its degradation products is at least 1 mg/L. The extrac the initiator in the Substrate pre-polymerization, or quantifi tion should be carried out for a sufficient time such that the 55 cation and/or detection of the initiator or its degradation prod change in concentration of the extract is not increasing more uct(s) in the Substrate post-polymerization may be accom than 5% per hour. Alternatively, extraction until the amount of plished by extraction followed by any of a range of analytical extracted material in a Subsequent extraction is less than 10% techniques. For example, quantifying and/or detecting the of that detected in the initial extraction, or until there is no amount of initiator or its degradation product(s) in the extract analytically significant increase in the cumulative extracted 60 can be accomplished using spectroscopy and chromatogra material levels detected. Exemplary extraction conditions phy; including, UV/VIS, FTIR, nuclear magnetic spectros include: 37° C. for 72 h; 50° C. for 72 h; 70° C. for 24 h; and copy, thin layer chromatography, gas chromatography, and 121° C. for 1 h. Exemplary extraction ratio includes 6 cm/ liquid chromatography. mL Surface area/volume and/or 0.2 g sample/mL. In some Monomers can be selected such that their reactivity ratios instances, complete dissolution of the Substrate may be 65 give alternating copolymers, periodic copolymers with a pre appropriate. Materials shall be cut into small pieces before specified ratio of each monomer, random copolymers, block extraction to enhance Submersion in the extract media, for copolymers or homopolymers. Inclusion of more than two US 9,096,703 B2 91 92 reactive groups on each monomer unit allows for the forma tion or Suspension of the monomer or monomers to be poly tion of star polymers, dendrimers, regularly branched poly merized. The quantity of polymer introduced to the substrate mers, randomly branched polymers, and brush polymers. In can be controlled by changing the concentration of the poly general, the monomer may be selected from any of the mono mer in the Solvent Solution, Surface tension of the polymer mers disclosed herein. Thus, for example, the monomers may 5 Solution, polymerization temperature, pH of the polymer contain any of the pendant groups corresponding to Formulae Solution, polymerization solution agitation or flow condi ZI-1 to ZI-7. By way of further example, upon polymerization tions, by changing the amount of time the Substrate is allowed the monomers may provide the polymer with repeat units to be in the polymer Solution during one polymerization corresponding to any of Formula 1-12. In a preferred embodi period, and/or by repeating any number of polymerization ment, the monomers are miscible with the polymerization 10 periods as required. When utilizing multiple polymerization mixture solvent system. periods, the polymer(s) used in the Subsequent polymeriza In processes for modification of the surface of a hydropho tion periods can be the same as, different from, or a mixture bic substrate, a hydrophilic solvent system preferably is with the polymer(s) used in the previous polymerization employed. Aqueous solutions preferably are used as the Sol period. vent system, optionally containing ions or buffers, such as 15 Chain transfer agents can be added to the monomer Solu Sodium, ammonium, potassium, chloride, phosphate, or tion to mediate the graft-from radical polymerization reaction acetate. In processes for modifying hydrophilic Substrates, a kinetics. Chain transfer agents include, but are not limited to, hydrophobic solvent system preferably is used. In such pro molecules containing halocarbons, thiols, dithiocarbamates, cesses, the preferred media is an organic solvent, typically a trithiocarbonates, dithioesters, Xanthates, primary or second non-polar organic solvent, or a mixture thereof. Exemplary ary alcohols. Examples of chain transfer agents are bromot organic solvents include one or more of toluene, hexane, richloromethane, 4-methylbenzenethiol, benzyl alcohol, cyclohexane, benzene, Xylene, tetrahydrofuran, and aliphatic methanol, ethanol, ethyleneglycol, glycerol, and isopropanol. alcohols. In a preferred embodiment, the solvent system does In one embodiment the radical polymerization graftings are not swell the substrate (or at least that portion of the substrate mediated using 2.2.6.6-tetramethylpiperidinie-1-oxyl from which the polymer will be grafted) by more than 25% by 25 (TEMPO). In one embodiment the radical polymerization Volume. For example, in one Such embodiment, the Solvent graftings are mediated using reversible addition fragmenta system does not swell the substrate (or at least that portion of tion transfer (RAFT) agents. Examples of RAFT agents the substrate from which the polymer will be grafted) by more include 2-(Dodecylthiocarbonothioylthio)-2-methylpropi than 10% by volume. In a preferred embodiment, the solvent onic acid, 2-Cyano-2-propylbenzodithioate, 2-Cyano-2-pro system does not swell the substrate (or at least that portion of 30 pyl dodecyl trithiocarbonate, 4-Cyano-4-(phenylcarbono the substrate from which the polymer will be grafted) by more thioylthio)pentanoic acid, 4-Cyano-4- than 5% by volume. In one embodiment, the solvent system (dodecylsulfanylthiocarbonyl)sulfanylpentanoic acid, Bis may even shrink the substrate (or at least that portion of the (dodecylsulfanylthiocarbonyl)disulfide, Bis(thiobenzoyl) substrate from which the polymer will be grafted). disulfide, Cyanomethyl dodecyl trithiocarbonate, In one particularly preferred embodiment, the non-fouling 35 Cyanomethyl methyl(phenyl)carbamodithioate, and their polymeric materials are grafted from the Substrate by chain analogues and derivatives growth addition polymerization. The polymerization condi Oxygen can act as an inhibitor in free radical polymeriza tions described herein are generally mild compared to other tion as it can react quickly with the free radicals generated by methods of polymerization and thus do not significantly alter the initiator to form stable radical species, which in turn can the mechanical properties, flexibility, or dimensional proper 40 react with other radical species to form unreactive species ties of the underlying substrate. In one preferred embodiment, which terminate the polymerization. Therefore, creating an for example, polymerization is carried out at a temperature oxygen-free environment by degassing with nitrogen or not in excess of 60°C. The polymerization may be carried out argon or vacuum is typically used to remove oxygen before over a relatively wide pH range, e.g., about 0-10. In one and during polymerization. However, for certain embodi embodiment, the polymerization reaction is carried out at a 45 ments, it would preferable not to require Such degassing steps pH of about 2-8. For example, when DCP and ferrous glu in commercial production. In one preferred embodiment, the conate are used as redox pair, the polymerization reaction polymerization method is other than ATRP, which typically may be carried out at a pH of about 6-8. By way of further requires stringent control of oxygen levels that may be diffi example, when benzoyl peroxide and ferrous gluconate are cult to achieve during manufacturing. used as redox pair, the polymerization reaction may be carried 50 Alternatively, oxygen in the system can be minimized by out at a pH of about 4-6. By way of further example, when filling the reactor with the reaction mixtures thus physically O.O-t-Butyl-O-(2-ethylhexyl)mono-peroxycarbonate displacing the oxygen in the reactor. In another embodiment, (“TBEC’) and ferrous gluconate are used as redox pair, the reagents which scavenge oxygen can be added to the reaction polymerization reaction may be carried out at a pH of about mixture. Suitable oxygen-Scavenging reagents include, but 5-7. 55 are not limited to, Sodium (meta) periodate, riboflavin, and Examples of radical polymerization processes include, but ascorbic acid. These agents may improve the efficacy of the are not limited to, UV, thermal, and redox initiated processes. resulting polymer if the polymerization does not employ an In particular embodiments, the polymer is grafted from the inert atmosphere. Substrate, by first incorporating one or more initiators, such as In addition to monomer and a solvent system, the polymer an ultraviolet (UV), thermal, or redox initiator into the sub 60 ization mixture may optionally containa free radical inhibitor strate and initiating polymerization of one or more monomers to encourage Surface grafting. Without being bound to any from the surface. Preferably, the initiator is incorporated into particular theory, it is presently believed that the addition of a the substrate by imbibing the substrate with initiator or coat free radical inhibitor, including, hydroquinone, hydro ing the Substrate with a layer, e.g., an undercoating layer quinone monomethyl ether, phenothiazine, 3.7-bis(dimethy (sometimes referred to herein as the co-deposited layer), 65 lamino)phenazathionium chloride, triethylene diamine, t-bu comprising the initiator. The polymerization is typically ini tylcatechol, butylated hydroxytoluene, and 4-t-butylphenol tiated by exposing the initiator-imbibed substrate with a solu to the grafting solution decreases solution polymerization, US 9,096,703 B2 93 94 thereby allowing more monomer to be available for grafting bonate, Cumene hydroperoxide, Cyclohexanone peroxide, at or near the Substrate surface/polymerization mixture inter Dicumyl peroxide, Lauroyl peroxide, 2.4-Pentanedione per face. oxide, Peracetic acid, Potassium persulfate. Plasticizers can be incorporated into the grafted polymer at The temperature to which the solution is heated is depen any time during and/or Subsequent to Surface polymerization. dent, among other things, on the monomer and/or the initiator, In the preferred embodiment, a hydrophilic plasticizer (such and/or the substrate. Examples of thermal radical initiators as citrated esters, ethylene glycol, propylene glycol, and/or include, but are not limited to, azo-compounds such as azo polyethylene glycol (<2000 MI) is incorporated into the bisisobutyronitrile (AIBN) and 1,1'-Azobis(cyclohexanecar grafted polymer in a post-polymerization aqueous wash bonitrile) (ABCN). Preferable grafting temperatures are near period. 10 the 10 hour T'/2 of the initiator selected. The graft-from radi i. UV Initiators cal polymerization reaction can be thermally quenched by In one embodiment, the initiator is an ultraviolet (UV) heating beyond the initiators half life. initiator. The substrate and initiator are typically placed into iii. Redox Initiators an aqueous, degassed, Solution containing a Zwitterionic In another embodiment, a redox initiator system is used to monomer and exposed to UV light, initiating the radical poly 15 initiate polymerization from the surface of the substrate. The merization. In one exemplary embodiment, the UV light has redox initiator System typically includes a pair of initiators: a peak wavelength of 365 nm, generated by a 100 WUV. an oxidant and a reducing agent. The redox chemistry Examples of UV radical initiators include, but are not described herein can be modified to prepare non-fouling limited to. 1-Hydroxycyclohexyl phenyl ketone, 2.2-Di polymeric materials, for example, Such as Zwitterionic poly ethoxyacetophenone, 2-Benzyl-2-(dimethylamino)-4'-mor meric materials. Redox initiation is regarded as a one-elec pholinobutyrophenone, 2-Hydroxy-2-methylpropiophe tron transfer reaction to effectively generate free radicals none, 2-Hydroxy-4-(2-hydroxyethoxy)-2- under mild conditions. Suitable oxidants include, but are not methylpropiophenone, 2-Methyl-4-(methylthio)-2- limited to, peroxide, hydroperoxide, persulfates, peroxycar morpholinopropiophenone, 3'-Hydroxyacetophenone, bonates, peroxydisulfates, peroxydiphosphate, permangan 4'-Ethoxyacetophenone, 4-Hydroxyacetophenone, 4'-Phe 25 ate, salts of metals such as Mn(III), Ce(IV), V(V), Co(III), noxyacetophenone, 4'-tert-Butyl-2',6'-dimethylacetophe Cr(VI) and Fe(III). none, Diphenyl(2,4,6-trimethylbenzoyl)phosphine oxide/2- Suitable reducing agents include, but are not limited to, hydroxy-2-methylpropiophenone, 2,2-Dimethoxy-2- metal salts such as Fe(II), Cr(II), V(II), Ti(III), Cu(II), and phenylacetophenone, 4,4'-Dimethoxybenzoin, 4,4'- Ag(I) salts, and oxyacids of Sulfur, hydroxyacids, alcohols, Dimethylbenzil, Benzoin ethyl ether, Benzoin isobutyl ether, 30 thiols, ketones, aldehydes, amine, and amides. For example, Benzoin methyl ether, Benzoin, 2-Methylbenzophenone, 3,4- in some embodiments, the reducing agent is an iron(II) salt, Dimethylbenzophenone, 3-Hydroxybenzophenone, 3-Meth such as iron(II) L-ascorbate, ferrous sulfate, iron(II) acetate, ylbenzophenone, 4,4'-Bis(diethylamino)benzophenone, 4,4'- iron(II) acetylacetonate, iron(II)ethylenediammonium Sul Dihydroxybenzophenone, 4,4'-Bis2-(1-propenyl)phenoxy fate, iron(II) gluconate, iron(II) lactate, iron(II) oxalate, or benzophenone, 4-(Diethylamino)benzophenone, 35 iron(II) sulfate. 4-Benzoylbiphenyl, 4-Hydroxybenzophenone, 4-Methyl Polymerization can be initiated by radicals formed directly benzophenone, Benzophenone-3,3,4,4-tetracarboxylic from the redox reaction and/or by macroradicals formed by dianhydride, Benzophenone, Methyl benzoylformate, the abstraction of a hydrogenatom from the substrate by the Michler's ketone, Sulfoniums, iodiums, 2-(4-Methoxy transient radicals formed during the redox reaction. styryl)-4,6-bis(trichloromethyl)-1,3,5-triazine, Diphenyli 40 In one embodiment, the substrate is coated with a under odonium p-toluenesulfonate, N-Hydroxy-5-norbornene-2,3- coating coating and the non-fouling material is grafted from dicarboximide perfluoro-1-butanesulfonate, the undercoating layer by redox polymerization. The under N-Hydroxynaphthalimide triflate, 2-tert-Butylan coating coating contains oxidants or reducing agents. In a thraquinone, 9,10-Phenanthrenequinone, Anthraquinone-2- preferred embodiment, the undercoating layer contains one Sulfonic acid sodium salt monohydrate, Camphorquinone, 45 or more reducing agents, such as acids, alcohol, thiols, Diphenyl(2,4,6-trimethylbenzoyl)phosphine oxide, 10-Me ketones, aldehydes, amines and amides. An oxidant is used to thylphenothiazine, thioxanthones, and IRGRCURE 2959. react with one or more functional groups of the undercoating ii. Thermal Initiators layer to form radicals which initiate the graft-from polymer In another embodiment a heat activated (thermal) initiator ization. is used, in place of the UV initiator described above, and the 50 In a particular embodiment, the undercoating layer is a graft-from polymerization is initiated by heating the aqueous copolymer with pendant groups of aliphatic chains contain monomer Solution temperature to a desired temperature and ing silanol and/or hydroxyl groups. Such materials can be holding the temperature constant until the desired degree of used to form a undercoating layer on polymeric Substrates, polymerization is achieved. Such as polyurethane (PU). An oxidant, such as a salt of Suitable thermal initiators include, but are not limited to, 55 Ce(IV), reacts with the hydroxyl group under mild conditions tert-Amyl peroxybenzoate, 4.4-Azobis(4-cyanovaleric acid), to form hydroxyl radicals in the undercoating layer to grow 2,2'-AZobis(2-carboxyethyl)-2-methylpropionamidine. the Zwitterionic polymers. 2,2'-Azobis(4-methoxy-2,3-dimethylvaleronitrile), 1,1'- In still another embodiment, a pair of peroxides and metal Azobis(cyclohexanecarbonitrile), 2,2'-Azobisisobutyroni salts (such as Fe(II) as used in the Fenton Reaction) is used in trile (AIBN), Benzoyl peroxide, 2.2-Bis(tert-butylperoxy) 60 the redox polymerization to graft Zwitterionic polymers from butane, 1,1-Bis(tert-butylperoxy)cyclohexane, 2.5-Bis(tert polymers such as polyurethane. Peroxides for use in the redox butylperoxy)-2,5-dimethylhexane, 2.5-Bis(tert polymerization include diacyl peroxides, dialkyl peroxides, Butylperoxy)-2,5-dimethyl-3-hexyne, Bis(1-(tert diperoxyketals, hydroperoxides, ketone peroxides, peroxydi butylperoxy)-1-methylethyl)benzene, 1,1-Bis(tert carbonates, and peroxyesters. Exemplary diacyl peroxides butylperoxy)-3,3,5-trimethylcyclohexane, tert-Butyl 65 include decanoyl peroxide, lauroyl peroxide. Succinic acid hydroperoxide, tert-Butyl peracetate, tert-Butyl peroxide, peroxide, and benzoyl peroxide, Exemplary dialkyl peroX tert-Butyl peroxybenzoate, tert-Butylperoxy isopropyl car ides include dicumyl peroxide, 2,5-di(t-butylperoxy)-2,5- US 9,096,703 B2 95 96 dimethylhexane, t-butyl cumyl peroxide, a.a'-bis(t-butylper aZobis(4-cyanovaleric acid), or 1, 1'-Azobis(cyclohexanecar oxy)diisopropylbenzene mixture of isomers, di(t-amyl) bonitrile), 2,2'-Azobisisobutyronitrile (AIBN) and (ii) the peroxide, di(t-butyl)peroxide and 2,5-di(t-butylperoxy)-2,5- hydrophilic reducing agent is Fe", Cr'', V'", Ti", Co", dimethyl-3-hexyne. Exemplary diperoxyketals include 1,1- Cu", oran amine; transition metalion complexes, e.g., copper di(t-butylperoxy)-3,3,5-trimethylcyclohexane, 1,1-di(t- (II) acetylacetonate, HSO, SO, SO, or SO’. Exem butylperoxy)cyclohexane, 1,1-di(t-amylperoxy) plary combinations include any of the aforementioned per cyclohexane, n-butyl 4,4-di(t-butylperoxy)valerate, ethyl oxides and Fe". In some preferred embodiments, benzoyl 3.3-di-(t-amylperoxy)butanoate and ethyl 3,3-di-(t-butylper peroxide, dicumyl peroxide, or O,O-t-Butyl-O-(2-ethyl oxy)butyrate. Exemplary hydroperoxides include cumene hexyl)mono-peroxycarbonate are used in combination with hydroperoxide and t-butyl hydroperoxide. Exemplary ketone 10 Fe2. peroxides include methyl ethyl ketone peroxide mixture and In an alternative embodiment, the hydrophilic-hydropho 2.4-pentanedione peroxide. Exemplary peroxydicarbonates bic redox pair is a hydrophilic oxidizing agent/hydrophobic include di(n-propyl)peroxydicarbonate, di(sec-butyl)per reducing agent pair wherein (i) the hydrophilic oxidizing oxydicarbonate, and di(2-ethylhexyl)peroxydicarbonate. agentisperacetic acid, a persulfate Such as potassium persul Exemplary peroxyesters include 3-hydroxy-1,1-dimethylbu 15 fate, Fe", CIO, HO, Ce", V, Cr', or Mn, or their tyl peroxyneodecanoate alpha-cumyl peroxyneodecanoate, combinations; and (ii) the hydrophobic reducing agent is an t-amyl peroxyneodecanoate, t-butyl peroxyneodecanoate, alcohol, carboxylic acid, amine, or a boronalkyl or their com t-amyl peroxypivalate, t-butyl peroxypivalate, 2,5-di(2-eth binations. ylhexanoylperoxy)-2,5-dimethylhexane, t-amyl peroxy-2- Other Suitable redox Systems include (1) organic-inorganic ethyl hexanoate, t-butyl peroxy-2-ethyl hexanoate, t-amyl redox pairs, such as oxidation of an alcohol by Ce", V", peroxyacetate, t-butyl peroxyacetate, t-butyl peroxyacetate, Cr", Mn"; (2) monomers which can act as a component of t-butyl peroxybenzoate, OO-(t-amyl) O-(2-ethylhexyl) the redox pair, such as thiosulfate plus acrylamide, thiosulfate monoperoxycarbonate, OO-(t-butyl)-O-isopropyl monoper plus methacrylic acid, and N,N-dimethylaniline plus methyl oxycarbonate, OO-(t-butyl)-O-(2-ethylhexyl)monoperoxy methacrylate, and (5) boronalkyl-oxygen systems. carbonate, polyether poly-t-butylperoxy carbonate, and 25 iv. Exemplary Initiators t-butyl peroxy-3.5.5-trimethylhexanoate. Exemplary initiators include, but are not limited to, diacyl Any of the aforementioned peroxides Such as benzoyl per peroxides such as benzoyl peroxide, dichlorobenzoyl peroX oxide, lauroyl peroxide, hydrogen peroxide, or dicumyl per ide, dilauroyl peroxide, didecanoyl peroxide, diacetyl peroX oxide are imbibed into the polymer such as polyurethane by ide Succinic acid peroxide, disuccinic peroxide and di(3.5.5- dipping the polymer into a peroxide Solution in an organic 30 trimethylhexanoyl)peroxide. In a preferred embodiment, the solvent for a predetermined period of time and dried. The diacyl peroxide is an aromatic diacyl peroxide, such as ben peroxide containing polymer is put into a solution of mono Zoyl peroxide. mer. The redox polymerization is initiated by the addition of Other exemplary initiators include, but are not limited to, a reducing agent, for example salts of Fe(II), such as Fe(II) peroxydicarbonates such as diethyl peroxydicarbonate, di-n- chloride, Fe(II) sulfate, ammonium Fe(II) sulfate, or Fe(II) 35 butyl peroxydicarbonate, diisobutyl peroxydicarbonate, di-4- gluconate, at room temperature or elevated temperature, to tert-butylcyclohexyl peroxydicarbonate, di-sec-butyl per the monomer Solution. oxydicarbonate, di-2-ethylhexyl peroxydicarbonate, di-n- For modifying the Surface of an article and/or Surface graft propyl peroxydicarbonate and diisopropyl polymerization, it has been found particularly useful to use peroxydicarbonate; peroxyesters, such as t-butyl perneode hydrophobic-hydrophilic redox initiator pairs. For example, 40 canoate, t-butyl and t-amyl peroxy 2-ethylhexanoate, and in one embodiment the hydrophobic member of a hydropho t-butyl peroxybenzoate; monoperoxycarbonates based on bic-hydrophilic redox initiator pair is incorporated into a t-butyl and t-amyl monoperoxy 2-ethylhexylcarbonates; per hydrophobic substrate as previously described. The substrate Sulfates. Such as potassium persulfate, ammonium persulfate, Surface is then treated with an aqueous polymerization mix and sodium persulfate; cumene hydroxide, tert-butyl hydro ture containing monomers, typically hydrophilic monomers, 45 peroxide, di(tert-amyl) peroxide, tert-butyl peroxide, 2.5-Bis and the hydrophilic member of the redox pair. This method (tert-butylperoxy)-2,5-dimethylhexane, 1,1-Bis(tert-butylp offers particular advantages when polymers are being grafted eroxy)-3,3,5-trimethylcyclohexane: 1,1-Bis(tert from components having exposed external and internal Sur amylperoxy)cyclohexane, 1,1-Bis(tert-butylperoxy)-3,3,5- faces to be modified (Such as catheters) and any Substrate that trimethylcyclohexane, 1,1-Bis(tert-butylperoxy) cannot readily be exposed to light. Additionally, Such a sys 50 cyclohexane, 2.2-Bis(tert-butylperoxy)butane, 2,4- tem tends to minimize the extent of non graft polymerization Pentanedione peroxide, 2.5-Bis(tert-butylperoxy)-2,5- in the bulk polymerization mixture away from the polymer dimethylhexane, 2,5-Di(tert-butylperoxy)-2,5-dimethyl-3- ization mixture/substrate Surface interface. hexyne, 2-Butanone peroxide, cumene hydroperoxide, In a preferred embodiment, the hydrophilic-hydrophobic di-tert-amyl peroxide, dicumyl peroxide, lauroyl peroxide, redox pair is a hydrophobic oxidizing agent/hydrophilic 55 tert-butyl peracetate, tert-butyl peroxide, tert-butyl peroxy reducing agent pair wherein (i) the hydrophobic oxidizing benzoate, tert-butylperoxy 2-ethylhexyl carbonate, tert-Bu agent is tert-amyl peroxybenzoate, O.O-t-Butyl-O-(2-ethyl tylperoxy isopropyl carbonate, 4-nitro-benzenecarboper hexyl)mono-peroxycarbonate, benzoyl peroxide, 2.2-bis oXoic acid t-butyl ester, cyclohexanone peroxide, (tert-butylperoxy)butane, 1,1-bis(tert-butylperoxy)cyclohex (methylperoxy) (diphenyl)methylbenzene, bis(t-butylcy ane, 2.5-bis(tert-butylperoxy)-2,5-dimethylhexane, 2.5-Bis 60 clohexyl)peroxydicarbonate, and 2,4,6-triphenylphenoxyl (tert-Butylperoxy)-2,5-dimethyl-3-hexyne, bis(1-(tert dimer. butylperoxy)-1-methylethyl)benzene, 1,1-bis(tert For Substrates requiring coating on both internal and exter butylperoxy)-3,3,5-trimethylcyclohexane, tert-butyl nal Surfaces, additional considerations are required for initi hydroperoxide, tert-butyl peracetate, tert-butyl peroxide, tert ating polymerization. Thermal initiators can be used; how butyl peroxybenzoate, tert-butylperoxy isopropyl carbonate, 65 ever, the elevated temperature typically required can cumene hydroperoxide, cyclohexanone peroxide, dicumyl adversely affect the substrate material. UV based approaches peroxide, lauroyl peroxide, 2.4-pentanedione peroxide, 4.4- must be designed such that they can penetrate through the US 9,096,703 B2 97 98 material or can be applied intralumenally, for instance from a Suitable devices include, but are not limited to surgical, medi fiber optic source threaded into the lumen. This may be cal or dental instruments, blood oxygenators, pumps, tubing, achieved by selecting a photoactive initiator which is labile at wiring, electrodes, contraceptive devices, feminine hygiene a UV wavelength not absorbed by the substrate polymer. products, endoscopes, grafts, stents, pacemakers, implant Generally, lower wavelength UV irradiation is less absorbed 5 able cardioverter-defibrillators, cardiac resynchronization and penetrates more readily than higher wavelength UV. therapy devices, Ventricular assist devices, heart valves, cath In contrast, redox chemistries generally do not require a eters (including vascular, urinary, neurological, peritoneal, direct line of sight to a light Source to initiate polymerization interventional, etc.), shunts, wound drains, dialysis mem branes, infusion ports, cochlear implants, endotracheal tubes, since polymerization is not initiated photolytically and there guide wires, fluid collection bags, sensors, wound treatments fore may be advantageous for coating Substrates that have one 10 (dressings, bandages, Sutures, cell scaffolds, bone cements, or more surfaces that are difficult to expose to the UV source, particles), ophthalmic devices, orthopedic devices (hip Such as catheter lumens. Further, redox polymerization typi implants, knee implants, spinal implants, screws, plates, riv cally can be done at low temperatures, for example less than ets, rods, intramedullary nails, bone cements, artificial ten 60° C., less than 55°C., less than 50° C., less than 45° C., less dons, and other prosthetics or fracture repair devices), dental than 40° C., less than 35° C., or less than 30° C. 15 implants, breast implants, penile implants, maxillofacial The graft-from polymerization can propagate through a implants, cosmetic implants, valves, appliances, scaffolding, cationic or anionic reaction, where the Substrate Surface acts Suturing material, needles, hernia repair meshes, tension-free as the cation oranion initiator or a cationic oranionic initiator vaginal tape and vaginal slings, tissue regeneration or cell is immobilized on the Substrate and the monomer contains a culture devices, or other medical devices used within or in reactive olefin. Examples of anionic polymerization are contact with the body or any portion of any of these. Prefer anionic ring opening, as in the case of synthesizing polyca ably, the non-fouling coating herein does not significantly prolactone or polycaprolactam, where the polymerization adversely affect the desired physical properties of the device proceeds through a lactone or lactam moiety in a ring struc including, but not limited to, flexibility, durability, kink resis ture containing a pendant Zwitterion group. Alternatively, an tance, abrasion resistance, thermal and electrical conductiv organic ring containing one or more units of unsaturation and 25 ity, tensile strength, hardness, and burst pressure. a pendant Zwitterionic group are polymerized. In one embodi In one embodiment, the Substrate is a vascularly inserted catheter Such as a peripherally inserted central catheter ment a pendant olefin is included in the monomer unit and is (PICC), central venous catheter (CVC) or hemodialysis cath used for crosslinking, Such as in ring opening metathesis eter, venous valves, punctual plugs, and intra-ocular devices polymerization (ROMP). and implants. Methods of Use 30 In another embodiment, the Substrate is a vascularly The materials described above may be in the form of a inserted catheter formed from a medical grade polyurtheth medical device or other article to which the non-fouling mate ane or CARBOTHANER) or formed from a material coated rial is grafted. Suitable devices include, but are not limited to, with a medical grade polyurethane. Surgical, medical or dental instruments, ophthalmic devices, In one specific embodiment, the catheter comprises an wound treatments (bandages, Sutures, cell scaffolds, bone 35 elongated catheter body containing multiple lumens. For cements, particles), appliances, implants, scaffolding, Sutur example, the catheter may be a double-lumen or a triple ing material, valves, pacemaker, stents, catheters, rods, lumen catheter. The lumens may be coaxial or side-by-side. In implants, fracture fixation devices, pumps, tubing, wiring, one exemplary embodiment, the catheter body has two side electrodes, contraceptive devices, feminine hygiene prod by-side lumens, each having a "D' shape and the catheter ucts, endoscopes, wound dressings and other devices, which 40 body has a length that is greater than 5 cm; typically the come into contact with tissue, especially human tissue. catheter body of such catheters have a length of at least 11 cm. In one embodiment, the non-fouling materials are grafted In one particularly preferred embodiment, the catheter body directly from a fibrous material, incorporated into a fibrous is a medical-grade polycarbonate-based aliphatic and aro material or grafted indirectly from a fibrous material (e.g., matic polyurethane. coated on a different Surface coating). These include wound 45 The non-fouling materials can also be added to paints and dressings, bandages, gauze, tape, pads, sponges, including other coatings and filters to prevent mildew, bacterial con tamination, and in other applications where it is desirable to woven and non-woven sponges and those designed specifi prevent fouling. Such as marine applications (ship hull coat cally for dental or ophthalmic surgeries (See, e.g., U.S. Pat. ings), contact lenses, dental implants, coatings for in vivo Nos. 4,098,728; 4,211,227; 4,636,208; 5,180,375; and 6,711, sensors, devices for separations, such as membranes for 879), paper or polymeric materials used as Surgical drapes, 50 microbial Suspension, biomolecule separation, protein frac disposable diapers, tapes, bandages, feminine products, tionation, cell separation, waste water treatment, bioreactors, Sutures, and other fibrous materials. and food processing. Fibrous materials are also useful in cell culture and tissue Other applications include the treatment of fibers, particu engineering devices. Bacterial and fungal contamination is a lates and films for applications in textiles, additives, electric/ major problem in eukaryotic cell culture and this provides a 55 optical appliances, carbon nanotubes, packaging materials safe and effective way to minimize or eliminate contamina and colorants/inks. tion of the cultures, Having described the invention in detail, it will be apparent The non-fouling agents are also readily bound to particles, that modifications and variations are possible without depart including nanoparticles, microparticles, millimeter beads, or ing from the scope of the invention defined in the appended formed into micelles, that have uses in a variety of applica 60 claims. Furthermore, it should be appreciated that all tions including cell culture, as mentioned above, and drug examples in the present disclosure are provided as non-lim delivery. Non-fouling, biocompatible, polymeric micelles iting examples. would prevent protein denaturation preventing activation of the immune response allowing for a more stealthy delivery of EXAMPLES the desired therapeutic. 65 The non-fouling material can also be applied directly to, or The following non-limiting examples are provided to fur incorporated in, polymeric, metallic, or ceramic Substrates. ther illustrate the present invention. It should be appreciated US 9,096,703 B2 99 100 by those of skill in the art that the techniques disclosed in the areas unoccupied by fibrinogen. The samples were rinsed, examples that follow represent approaches the inventors have transferred to clean wells, and incubated for 1 hour with 5.5 found function well in the practice of the invention, and thus ug/mL horseradish peroxidase conjugated anti-fibrinogen in can be considered to constitute examples of modes for its 10% (v/v) fetal bovine serum. Again the samples were rinsed 5 and transferred to clean wells with 0.1M phosphate-citrate practice. However, those of skill in the art should, in light of buffer containing 1 mg/mL chromogen of o-phenylenedi the present disclosure, appreciate that many changes can be amine and 0.02% (v/v) hydrogen peroxide. Incubating at 37° made in the specific embodiments that are disclosed and still C. for 20 minutes produces an enzyme-induced color reac obtain a like or similar result without departing from the spirit tion, which is terminated by the addition of 2.0Nsulfuric acid. and scope of the invention. The absorbance of light intensity was then be measured using Radio-Labeling Method for all Examples 10 a microplate reader to determine the protein adsorption rela The test samples are placed in a Suitable sized container, tive to controls (i.e., reference substrates). For mixed protein which may be a 96-well manifold, microcentrifuge tube, or Solutions, such as whole plasma, Surface plasmon resonance other container. The Volumes in the following are appropriate (SPR) or optical waveguide lightmode spectroscopy (OWLS) for a 96-well plate, but may be scaled to properly cover a can be utilized to measure surface protein adsorption without device being tested. The samples are sterilized with 70% 15 necessitating the use of individual antigens for each protein ethanol Solution for thirty minutes and the test groups run present in Solution. with an in per run of 3-4. The sample container is blocked with 20 mg/mL Bovine Serum Albumin in 1xEBS for 1 hour at 4 Example 3 C., followed by three rinses with 1xPBS before samples are added. The sample is exposed to 300 uL of a 70 ug/mL Polypropylene Substrates unlabeled fibrinogen solution containing 1.4 ug/mL I-125 radiolabeled fibrinogen. The samples is incubated for one Polypropylene coupons (0.4" thick, 1x0.5 cm) were imbibed with 5% BPO/toluene solution for 2 hours, and then hour at 37° C. and put on an orbital shaker at 150 RPM. The reacted with 20 wt % SBMA solution and 5 mM Fe(II) glu test solution is then removed and four 1-minute rinses with a conate in a 20 mL vial, at 40°C. for 5 hours. A determination 10 mMNaI and one 1-minute rinse with 1xPBS is performed. 25 of fibrinogen adsorption using the radiolabeling method The samples are be loaded into a gamma counter. The counter shows fibrinogen adsorption was reduced by 94+1% after measures the radioactivity in I-125 counts perminute for each grafting. Absolute adsorption offibrinogen was 14 ng/cm in sample and this data is used to calculate a percent reduction of a fibrinogen adsorption assay in which samples are incubated the non-fouling polymer layer samples Versus the reference for 60 minutes at 37°C. in 70 ug/mL fibrinogen derived from Substrates, that is the same or an otherwise functionally 30 human plasma. equivalent Substrate lacking the non-fouling polymer layer. The percent reduction is equal to: (1-non-fouling sample Example 4 CPM/Average CPM of the Reference Substrate)*100%. Silicone Substrates Example 1 35 Silicone tubes were imbibed with 5% dicumyl peroxide Ti Substrates (DCP)/heptanes solution for 2 hours and dried overnight. Then the tubes were reacted with 20 wt % SBMA solution and Titanium coupons (99.5%, 0.25 mm thick, 1 x0.5 cm) were 5 mM Fe(II) gluconate at 37° C. shaking at 100 rpm for 24 first washed with acetone under sonication for 10 min and 40 hours. A determination of fibrinogen adsorption using the dried in the air, then treated with piranha solution for 30 min. radiolabeling method shows fibrinogen adsorption was After clean and drying, the samples were coated with 1% reduced by 73-4% after grafting. polyurethane (Tecoflex)/THF solution with 0.1% BP and air-dried in dark for 30 min. Then coupons were dip-coated Example 5 for another three times and dried in dark over 3 nights. The 45 solution was flushed with nitrogen for 30 min before the MPC Monomer reaction, and reacted under a UV reactor for 14 h. The samples were rinsed with PBS. A determination of fibrinogen Polyurethane (Carbothane(R) rods were imbibed with 1 wt. adsorption using the radiolabeling method shows fibrinogen % TBEC/acetone solution for 3 hours. Redox reaction was adsorption was reduced 75%. 50 performed in 10% methacryloyloxyethyl phosphorylcholine (MPC) solution with 5 mM ferrous gluconate in a 20-mL vial. Example 2 The reaction was kept at 60° C. under shaking at 150 rpm overnight. Then the samples were washed with 20 mL PBS Polycarbonate Substrates three times with shaking at 150 rpm overnight. With adjust 55 ment made for non-modified ends of the sample, the absolute Polycarbonate central posts were imbibed with 10 wt.% adsorption of fibrinogen was 33 ng/cm on modified samples BP/acetone solution for 6 hours. The sample was kept in dark and the reduction from control was 96% using a fibrinogen at RT overnight. The samples were washed with water before adsorption assay in which samples are incubated for 60 min reaction. The reaction was performed in a reaction tube and utes at 37°C. in 70 g/mL fibrinogen derived from human 10 wt.% carboxybetaine acrylamide solution was added into 60 plasma. the tube. A 30-min degassing was applied by bubbling with argon before the reaction. The reaction was exposed to UV Example 6 light for 6 hours (peak wavelength of 365 nm, generated by a 100 W UV lamp). After washing with PBS, the samples Polyampholyte/Mixed Charge exhibited a fibrinogen deduction of 92.7% by ELISA. 65 In the ELISA assay, the samples were incubated for 90 Polyurethane (Carbothane(R) rods (3 cm) were imbibed minutes at 37°C. in 10% (v/v) fetal bovine serum to block the with 1 wt.% benzoyl peroxide (BPO)/acetone solution for 3 US 9,096,703 B2 101 102 hours. Redox reaction was performed in 10% mixed mono storage (rinsed in 1xEBS post reaction then air dried over mers (sulfopropyl methacrylate potassium salt (“SMP) and night and stored in sterilization bag) and put directly into 4% 2-(Methacryloyloxy)ethyltrimethylammonium chloride aqueous osmium tetroxide. All modified samples were cut methacrylate (“META)) with 5 mM ferrous gluconate in a from a large 25 cm treated tube. Two modified samples were 250 mL round bottom flask. The reaction was kept at 60° C. stained with osmium tetroxide while a third repetition was left under stirring for 5 hours. Then the samples were washed with unstained. Samples were left in osmium tetroxide overnight PBS three times. Fibrinogen reduction on the surfaces was and then rinsed thoroughly with distilled water. Any wet measured by the radiolabeling method and is reported in the stained samples were ethanol dehydrated (submerged for two following table. With adjustment made for non-modified ends minutes each in solutions of 50, 60, 70,80, 90 & 95% ethanol) 10 and then let air dry. of the sample, absolute adsorption of fibrinogen and reduc All of the samples were freeze fractured for imaging by tions from control are shown in the table below using a being Submerged in liquid nitrogen then cracked with an ultra fibrinogen adsorption assay in which samples are incubated microtome blade (Leica Ultracut UCT Ultramicrotome). One for 60 minutes at 37°C. in 70 ug/mL fibrinogen derived from half of the freeze fractured samples were put in distilled water human plasma. 15 while the other halfwere prepared for dry imaging under high vacuum. The dry samples were sputter coated in gold for 90 Fibrinogen Fibrinogen seconds using a sputter coater (Cressington 208HR) and then Sample description Adsorption (ng/cm) Reduction imaged under high vacuum at 5 kV using an SE2 detector under a Scanning Electron Microscope (SEM) (Supra55VP Carbo-Ba1% 91 89.5% BPO.10%.SMP:META = 40:60 FESEM, Zeiss). The thickness of dried coating was measured Carbo-Ba1% 92 88.3% along the interfaces (Ldry). BPO.10%.SMP:META = 45:SS The wet samples were first imaged after soaking in water Carbo-Ba1% 85 89 2% for approximately 4 hours. They were submerged wet into BPO.10%.SMP:META - SOSO liquid nitrogen and then fixed to a cold stage setto -8°C. The Carbo-Ba1% 81 90.3% BPO.10%.SMP:METASS:45 25 samples were then imaged using a VPSE detector at the Carbo-Ba1% 59 93 6% highest resolvable humidity (approximately 26% or 81 Pa) BPO.10%.SMP:META - 60:40 under a Scanning Electron Microscope (SEM) (Supra55VP FESEM, Zeiss) with an Environmental Scanning Electron Microscope (E-SEM) (Zeiss EVO 55). When the samples Example 7 30 were warmed and removed from the cold stage it was ensured that they were still wet. The final measurements were taken Surface Contact Angle Measurement after the samples were soaking in 1xRBS for at least 1 hr (before going into 1xPBS they were soaking in distilled water Flat silicone samples (Sylgard 184, Dow Corning, MI) and overnight). Immediately before imaging, the samples were silicone tubes (TYGON3350, Cole-Parmer Instrument Com 35 rinsed quickly with water to remove any Surface salts and then pany, IL) were modified with SBMA as described in example Submerged in liquid nitrogen and fixed to the cold stage. The 4. Additionally, flat polyurethane films (Tecoflex, SG93A, thickness of hydrated Samples was measured along the inter Lubrizol) were modified with SBMA as described in example faces (Lhyd). 4. The samples were first soaked with pure ethanol for 5 The linear swellability of the coating was calculated by, minutes and washed with PBS for three times. The samples 40 were then soaked within PBS (150 mM, pH 7.4) for 24 hours The results of the thickness and the linear swellability are and washed three times with purified water. Then the samples listed in the following table. were dried under a flow of air for 5 min before testing. The contact angle was measured using a video contact angle sys tem (VCA 2000, AST Inc.). A drop of purified water (1 uL) 45 was deposited on the test surface. Then the shape of the Thickness + SD (nm) LSW droplet was taken by a microscope with a CCD camera, and L (n = 7) 668 170 the contact angle was determined using VCA Optima XE. L. (H2O) (n = 7) 8O1 287 20% Following are the resulting contact angle measurements. L (PBS) (n = 12) 905 - 262 36% 50

Description Contact Angle (degree) Example 9 SBMA modified flat silicone 1O.S Unmodified flat silicone 117.9 Initiator Penetrating Measurement SBMA modified silicone tube 23.4 55 Unmodified silicone tube 95.7 Polyurethane rod (10 French Carbothane(R) rods with 20% SBMA modified Tecoflex film 5.3 BaSO) imbibed with different initiators (10% dicumyl per Unmodified Tecoflex film 73.3 oxide and 1% benzoyl peroxide) after reaction were cross sectioned with a razorblade. The cross-sectional Surface was 60 analyzed under a Nicolet iN-10 IR microscope (Thermo Sci Example 8 entific) FTIR. The IR spectrum from an attenuated total reflection (ATR) tip was used to detect the penetrating depth Coating Dry and Humidified Thickness of imbibed initiator from the edge of the samples. The ATR Measurement FTIR spectra on the center of the polyurethane rod imbibed 65 with 10% dicumyl peroxide (“DCP) showed a strong peak at Polyurethane (Carbothane(R) with barium sulfate DD tube ca. 700 cm-1 (characteristic peak of DCP) and the ATR-FTIR samples (25 cm) for Swelling analysis were taken from dry spectra on the center of the polyurethane rod imbibed with 1% US 9,096,703 B2 103 104 benzoyl peroxide (“BPO') showed a strong peak at ca. 700 Example 13 cm (characteristic peak of BPO) Surface Roughness Example 10 5FR double D lumen tubing samples (3 cm in length) of Comparative Performance SBMA and OEGMA polyurethane (TecothaneR)-30% BaSO were imbibed with a solution of TBEC, washed, and dried. The imbibed samples were then modified with an aqueous solution of SBMA and Polyurethane (Carbothane(R) rods (3 cm) were imbibed iron(II) gluconate for 2 hours for some samples and for 3 with either 10 wt.% dicumyl peroxide (DCP)/acetone, or 1 wt hours for others, and washed with PBS. The estimated thick % O.O-t-Butyl-O-(2-ethylhexyl)mono-peroxycarbonate 10 ness by IR analysis, and the roughness as determined from (“TBEC')/acetone solution for 2 h. Modification was per LEXT confocal microscopy were determined and is shown in formed on the imbibed samples using 10% SBMA or 1% the table below. Roughness (R) was 0.259 um for the OEGMA with 5 mM ferrous gluconate. The reaction was kept unmodified, 0.162 um for the sample modified for 2 hours, at 60° C. under stirring for 5 hours. Then the samples were and 0.107 um for the sample modified for at 3 hours. Modi washed with PBS three times. 15 fication thickness was 677 nm for the sample modified for 2 The results depict a difference between the Standard Redox hours and 1218 nm for the sample modified for 3 hours. 10% SBMA and the 1% OEGMA monomer. The 1% OEGMA exhibited inferior performance (log reduc Modification Roughness tion=0.86) when tested in the mGDC E. coli as compared to Reaction thicness (Rd) the Standard Redox 10% SBMA (log reduction=2.24). Time (nm) (um) Control Not modified O.259 Example 11 (not modified) 2h 677 - 157 O.162 3h 1218 237 O.107 Protein Resistance 25

Catheter Walls Example 14 Catheters (613.7+1.8 mm body length) were imbibed with O.O-t-Butyl-O-(2-ethylhexyl)mono-peroxycarbonate 30 Residual Initiator Visualization (“TBEC’) and modified with SBMA monomer and Fe(II) reaction solution. The modified samples (Lots “A” and “B”) In the presence of an oxidant such as peroxide, iodide is were washed and dried. converted to iodine. The relative amount of peroxide can be qualitatively determined by treating a test article with an Protein resistance of the outside portion of the shaft of the iodide Salt solution and comparing the presence and/or inten samples was determined using the radiolabeling method and 35 sity of the resulting orange-brown iodine stain. Iodide solu the results appear in the following table. tions were prepared by adding 3-6 g NaI salt to 10 mL EtOH (reagent alcohol, 95%), Vortexing the mixture for 10 seconds, Estimated allowing the vortexed mixture to rest for 10 minutes to dis Modification solve the salt, and then filtering the mixture through a PTFE 40 Thickness by % filter (0.45um) to remove any undissolved particles. Each test Lot IR Reduction Std Dew Fg (ngi cm2) article (e.g., 1.5 cm in length) was soaked with, for example, A. 1654 299 87% 2% (n = 4) 47 1 mL of the ethanolic NaI solution prepared above in a test (n = 6) tube for one minute. Then, the test article was taken out and B 1630 - 201 90% 2% (n = 4) 37 dried by a flow of air for one minute to remove solvent. The (n = 5) 45 test article was then put in an 8 mL scintillation vial with cap tightened, and placed in a 60°C. oven for 45 minto develop the color on the surface. The presence of stain was then Example 12 recorded and could be compared with a yellow tone chart (ranging from 0 to 10) to get a darkness number. Contact Angle 50 Four sets of test articles were immersed in a solution of sodium iodide, rinsed, and heated in a 60° C. oven as Polyurethane (Tecothane(R)-30% BaSO-5 FR DD lumen described. The first set was untreated polyurethane (Teco catheters (562.2+0.9 mm body length) were imbibed with thane(R97A)-30% BaSO 5 FR double D lumen tubing. The O.O-t-Butyl-O-(2-ethylhexyl)mono-peroxycarbonate second set was polyurethane (TecothaneR)97A-30% BaSO (“TBEC’) and modified with SBMA monomer and Fe(II) 55 5 FR double D lumen tubing, imbibed with TBEC, but no reaction Solution. Contact angle of the outside portion of the surface modification was grown. The third and fourth sets shaft of these devices and for unmodified controls are shown were polyurethane (Tecothane 97A(R)-30% BaSO 5 FR below. double D lumen tubing, imbibed with TBEC and surface modified with SBMA. Iodine staining was not observed for 60 the first set, but was for the other three sets. Estimated Modification Contact Angle Example 15 Sample Thickness by IR (degrees) St Dew Unmodified catheter Not applicable 79.7 2.4 Residual Initiator Determination by Extraction Modified catheter 3589 (n = 1) 20.9 1.5 65 Test articles (polyurethane (Tecothane(R) 97A-30% BaSO 5 FR double D lumen tubing) were imbibed with US 9,096,703 B2 105 TBEC and surface modified with SBMA. The extraction con -continued ditions are summarized in the following table and the results demonstrate that the initiator was extracted with 95% ethanol. % TBEC Sample Composition St. Dew. Surface modified samples, O.OOS24 O.OO106 Extraction condition Iodine stain observed (yesino) with additional isopropanol wash* 95% Ethanol 37° C. for 2h O 70% Ethanol 37° C. for 2h yes This data represents the average of three replicate sample injections, 10% tween 37° C. for 2h yes **This data represents the average of four replicates of thee samples. PBS 37° C. for 2 yes 10 Example 18 Example 16 Surface Modification Thickness Variation Residual Initiator Determination by IR 15 Thrombogenicity of modified polyurethane (Tecothane(R) 97A)-30% BaSO5 FR double Dlumen tubing with different IR analysis of a 25 cm polyurethane (Carbothane(R)- SBMA modification thicknesses. BaSO 14 FR double lumen tube that was imbibed with (a) Tecothane-BaSO4 5 FR, DD lumen tubing, was dicumyl peroxide and surface modified with SBMA in the imbibed with TBEC and modified with SBMA from an iron presence of iron(II) gluconate, washed and dried, showed a (II) gluconate aqueous solution. Modifications of different peak at 699 cm indicating the presence of the cumyl group. grafted SBMA thickness on the outside of the tubing as esti mated by IR analysis, were obtained by adjusting the grafting Example 17 time. These are summarized in the table below. 25 (b) The in-vitro evaluation of catheters using the flow Residual Initiator Quantitation by GCMS model described herein, with the following distinctions: the ends of the catheter tubing were sealed with 5 minute epoxy, Polyurethane (Tecothane(R)-30% BaSO-5 FR DD lumen and 20 cm of catheter tubing body was inserted into the loop. catheters were imbibed with O.O-t-Butyl-O-(2-ethylhexyl) The flow rate was 200 mL/min for 60-90 minutes. The cath mono-peroxycarbonate (“TBEC) and some of the imbibed 30 eter tubing test articles were then rinsed with saline, photo samples were surface modified with SBMA monomer and graphed, and sectioned (1 cm at distal (tip), 2 cm at insertion), Fe(II) reaction solution. The imbibed only and surface modi and the adhered radio labeled platelets were counted using a fied samples were washed and dried. Some of the modified gamma counter. The results are Summarized in the table samples were washed with isopropanol prior to washing and below. drying. 35 Quantification of peroxides and their decomposition prod Thickness Average ucts: Quantification of TBEC (OO-tert-butyl O-(2-ethyl Estimation of Radiolabeled hexyl)monoperoxycarbonate), using GCMS. The quantity of SBMA Platelet TBEC was determined based on GCMS analysis of methan modification adherence olic sample extracts. The samples were cryogenically ground 40 from IR analysis (% Control) to a fine powder before extraction. A5 ml aliquot of methanol O 100 was added to approximately 370 mg of each sample. The 91 74 6.5 extraction was allowed to continue with gentle agitation. The 620-231 2 samples were filtered through a 0.2 Lum PTFE syringe filter 3353 726 1 prior to analysis by GCMS. Because the compound of interest 45 is prone to degradation, resulting in a series of decomposition products as observed by GCMS, quantitation was based on Example 19 ethyl hexanol, the major degradation product observed in analysis of TBEC standards (assuming there are no other Surface Roughness Evaluation Sources of ethylhexanol in the samples). A series of dilutions 50 was prepared of a TBEC standard in methanol and a calibra Polyurethane (Tecothane(R)-30% BaSO-5 FR DD lumen tion curve was prepared based on the ethylhexanol peak. catheters (562.2+0.9 mm body length) were imbibed with Concentration of TBEC was calculated based on the calibra O.O-t-Butyl-O-(2-ethylhexyl)mono-peroxycarbonate tion prepared and the percent composition in each sample was (“TBEC’) and modified with SBMA monomer and Fe(II) calculated using the following equation: % TBEC 55 reaction solution. The modified samples were washed and Composition concentration (mg/mL)x5 mL mass sample dried. The Surface roughness of the modified samples was extracted (mg)x100%. A Summary of the average percent compared to unmodified samples by LEXT Confocal Micro TBEC composition values calculated for the sample extracts Scope Imaging and Roughness Analysis. Samples were is provided in the table below. mounted on a glass slide by double-sided tape. Digital 3-D 60 images were taken with the Olympus LEXTOLS4000 laser confocal microscope (“LEXT) under an Olympus % TBEC MPLAPON 50x objective lens. The digital images taken in Sample Composition St. Dew. this way have a 256x256 um field area. The Z-direction Surface modified O.208 O.O2O repeatability for this LEXT machine has been certified by samples* 65 Olympus to be less than 0.012 um. To measure the roughness, Imbibed only samples* O3O4 O.007 at least three images were taken from each sample and the Rrms roughness calculated using a 9 um cut-off length. As US 9,096,703 B2 107 108 measured, the un-modified Substrate had an R, of 0.321 um, whereas the modified substrate had an R of 0.199 um. Formula 2 X3 What is claimed is: H 1. An article of manufacture comprising a polymeric Sub C - C strate having a Surface and a grafted polymer layer on the Substrate Surface, the grafted polymer layer having a global k average dry thickness of at least about 500 nanometers, the Substrate Surface and the grafted polymer layer, in combina wherein tion, constituting a modified Surface having a fibrinogen X is hydrogen, alkyl or substituted alkyl, and adsorption of less than about 125 ng/cm in a fibrinogen 10 X is a pendant group comprising an oxylated alkylene binding assay in which the modified surface is incubated for moiety, a Zwitterionic moiety, an anionic moiety, or a 60 minutes at 37° C. in a composition containing 70 ug/ml cationic moiety. fibrinogen derived from human plasma and 1.4 ug/ml 1-125 11. The article of claim 1 wherein the grafted polymer layer radiolabeled fibrinogen and a water contact angle of less than 40 degrees. 15 comprises a polymer having repeat units corresponding to 2. The article of claim 1, the article further comprising a Formula 4: Solvent extractable polymerization initiator or degradation product thereof. 3. The article of claim 1 wherein the grafted polymer layer Formula 4 has an global average dry thickness that is at least equal to the A B C D global average R. Surface roughness of the Substrate Sur X3 X3 X3 X3 face. H H H H 4. The article of claim 1 wherein the grafted polymer layer -1 t-C C C d is a carboxybetaine or a Sulfobetaine polymer. X4 5. The article of claim 1 wherein the grafted polymer layer 25 (O or NH) O (O or NH) SO (O or NH) SO has a standard deviation of the global average dry thickness that does not exceed 50% of the global average dry thickness pi of the grafted polymer layer. O Y 6. The article of claim 1 wherein the magnitude of the 30 2^3, N difference between the global average dry thickness of the g grafted polymer layer as determined by scanning electron p microscopy (SEM) and the global average humidified thick OY O G 29 ness of the grafted polymer layer as determined by environ G O11 || mental scanning electron microscopy (ESEM) is less than O 200% of the global average dry thickness. 35 7. The article of claim 1 wherein the grafted polymer layer wherein comprises a chain growth polymer. a is 0-1; 8. The article of claim 1 wherein the grafted polymer layer comprises a Zwitterionic polymer. b is 0-1; 40 c is 0-1; 9. The article of claim 1 wherein the grafted polymer layer d is 0-1; comprises a polymer having repeat units corresponding to m is 1-20 Formula 1 in and o are independently 0-11; p and q are independently 0-11; Formula 1 45 X is hydrogen, alkyl or substituted alkyl, X1 X3 X is OX', NX'X', SX', aryl, heteroaryl or acyl: C-C X' is hydrogen, hydrocarbyl, substituted hydrocarbyl, X2 X4 heterocyclo or acyl: 50 X' and X’ are independently hydrogen, hydrocarbyl, substituted hydrocarbyl or heterocyclo; and wherein X is hydrogen, hydrocarbyl or substituted hydrocarbyl, X' and X’ are independently hydrogen, hydrocarbyl, sub provided the sum of a, b, c and disgreater than 0 and X of stituted hydrocarbyl, heterocyclo, or substituted carbo repeat unit D differs from the corresponding pendant nyl, provided, however, X and X’ are not each selected 55 from the group consisting of aryl, heteroaryl, and het group of repeat units A, B and C. erosubstituted carbonyl, 12. The article of claim 1 wherein the grafted polymer layer X is hydrogen, alkyl or substituted alkyl, comprises a polymer having repeat units corresponding to X4 is OX40, NX41X42, N*X41X42x43, SX40, aryl, Formula 5. Formula 6, Formula 7, Formula 8, or Formula 9: heteroaryl or acyl, 60 X' is hydrogen, hydrocarbyl, substituted hydrocarbyl, heterocyclo or acyl, and Formula 5 X', X" and X’ are independently hydrogen, hydrocar byl, substituted hydrocarbyl or heterocyclo. 10. The article of claim 1 wherein the grafted polymer layer 65 comprises at least one repeat unit corresponding to Formula 2: US 9,096,703 B2 110 -continued X" is hydrogen, hydrocarbyl, substituted hydrocarbyl, Formula 6 heterocyclo or acyl, and X7 X' and X’ are independently hydrogen, hydrocarbyl, substituted hydrocarbyl or heterocyclo. X- e 13. The article according to claim 1, wherein the modified X8 surface has a fibrinogen adsorption of less than about 90 Formula 7 ng/cm in a fibrinogen binding assay in which the modified --X-HET-- surface is incubated for 60 minutes at 37°C. in a composition Formula 8 containing 70 ug/ml fibrinogen derived from human plasma X10 X13 10 and 1.4 ug/ml 1-125 radiolabeled fibrinogen. 14. The article according to claim 1, wherein the modified X9 X12 surface has a fibrinogen adsorption of less than about 70 XII X14 ng/cm in a fibrinogen binding assay in which the modified Formula 9 surface is incubated for 60 minutes at 37°C. in a composition O X7 15 containing 70 ug/ml fibrinogen derived from human plasma ----e.g and 1.4 ug/ml 1-125 radiolabeled fibrinogen. --O O L L4-H 15. The article according to claim 1, wherein the modified O X8 surface has a fibrinogen adsorption of less than about 50 O ng/cm in a fibrinogen binding assay in which the modified surface is incubated for 60 minutes at 37°C. in a composition wherein containing 70 ug/ml fibrinogen derived from human plasma HET is part of a heterocyclic structure, and 1.4 ug/ml 1-125 radiolabeled fibrinogen. X is hydrogen, alkyl or substituted alkyl, 16. The article according to claim 1, wherein the polymeric X is OX', NX'X', SX', aryl, heteroaryl or Substrate comprises polyurethane. acyl, 25 17. The article of claim 16 wherein the modified surface X is ester, anhydride, imide, amide, ether, thioether, has a water contact angle of less than 30 degrees. thioester, hydrocarbylene, substituted hydrocarbylene, 18. The article of claim 16 wherein the modified surface heterocyclo, urethane, or urea; has a water contact angle of less than 25 degrees. X is hydrocarbylene, substituted hydrocarbylene, hetero 19. The article of claim 16 wherein the modified surface cyclo, amide, anhydride, ester, imide, thioester, thioet 30 has a water contact angle of less than 20 degrees. her, urethane, or urea; 20. The article of claim 1 wherein the surface of the poly X7 is hydrogen, alkyl or substituted alkyl: meric substrate has a native water contact angle of at least 90 X is an anionic moiety; degrees. X is hydrocarbylene, substituted hydrocarbylene, hetero 21. The article of claim 20 wherein the modified surface cyclo, amide, anhydride, ester, imide, thioester, thioet 35 has a water contact angle of less than 30 degrees. her, urethane, or urea; 22. The article of claim 20 wherein the modified surface X' is hydrogen, alkyl or substituted alkyl: has a water contact angle of less than 25 degrees. X' is a cationic moiety; 23. The article of claim 20 wherein the modified surface X' is hydrocarbylene, substituted hydrocarbylene, hetero has a water contact angle of less than 20 degrees. cyclo, amide, anhydride, ester, imide, thioester, thioet 40 24. The article of claim 1 wherein the polymeric substrate her, urethane, or urea; comprises a radiopaque additive. X' is hydrogen, alkyl or substituted alkyl: 25. The article of claim 24 wherein the polymeric substrate X'' is an anionic moiety; comprises a radiopaque additive selected from the group con L' and L are independently hydrocarbylene, substituted sisting of barium sulfate, bismuth salts, gold foil, and tanta hydrocarbylene, heterocyclo, amide, anhydride, ester, 4 5 lum. imide, thioester, thioether, urethane, or urea; and