RATIONAL DESIGN AND SYNTHESIS OF BIODEGRADABLE POLYURETHANE BASED BIOMATERIALS by CANCAN XU Presented to the Faculty of the Graduate School of The University of Texas at Arlington in Partial Fulfillment of the Requirements for the Degree of DOCTOR OF PHILOSOPHY THE UNIVERSITY OF TEXAS AT ARLINGTON December 2018 Copyright © by Cancan Xu 2018 All Rights Reserved ii Acknowledgements I would like to express my deepest appreciation to my supervising professor, Dr. Yi Hong, who guides me with his profound knowledge and sharp insights and inspires me to keep going by his research passion and dedication to his career. I cannot find words to thank him for all the instructions and help in the past 5 years and in future. Without his guidance and support this dissertation woinuld not have been possible. I am sincerely grateful to my dissertation committee, Dr. Liping Tang, Dr. Baohong Yuan, Dr. Zui Pan, Dr. Jun Liao, and Dr. Yi Hong, for their great insight and help for me outlining and writing the dissertation. Also, I would like to acknowledge Dr. Liping Tang and Yihui Huang for their great assistant with all of my in vivo studies. Moreover, I would like to acknowledge Dr. Zui Pan and Xian Liu for their assistance with some of my biological experiments. I also sincerely thank Dr. Jun Liao for his patient guidance of my research and study and Dr. Xiaodan Shi for her assistance with my biomechanical testing. In addition, I would like to thank Dr. Alejandro Bugarin from Chemistry and Biochemistry department in the University of Texas at Arlington for his kind help with some of my chemical synthesis. Many thanks to my colleagues, Dr. Jinglei Wu, Danh Truong, Yihui Huang, Dr. Zi Wei, and Aneetta E Kuriakose, for their support, contributions, and encouragements during the past years. In addition, thanks to Dr. Baohong Yuan, Dr. Kytai T. Nguyen, Dr. Guohao Dai, Dr. Mario I. Romero-Ortega, Shuai Yu, Dr. Nikhil Pandey, Taylor B. Dorsey, and Sanjay Anand for working together on so many exciting projects. iii A special gratitude goes to all research grants for partial support of my research work listed below: Research Enhancement Program (Y.H.) of the University of Texas at Arlington, Beginning Grant-in-Aid 14BGIA20510066 (Y.H.) from the American Heart Association, Faculty Career Development (CAREER) award #1554835 (Y.H.) from the National Science Foundation, and R21HD090680 (Y.H.) from the National Institutes of Health. And finally, last but not the least, I am so grateful to my parents for their support and love. They always keep me going. Also, I would like to thank my friends for sharing my sorrows and happiness in the past years. iv Abstract RATIONAL DESIGN AND SYNTHESIS OF BIODEGRADABLE POLYURETHANE BASED BIOMATERIALS Cancan Xu, PhD The University of Texas at Arlington, 2018 Supervising Professor: Yi Hong Biodegradable polyurethanes have been applied in biomedical field since 1960s. In recent years, biodegradable polyurethanes have been widely investigated for tissue repair and regeneration because of their good mechanical properties, elasticity, biodegradability, biocompatibility and processability. Compared with some commonly used biodegradable polymers, such as poly(glycolide) (PGA), poly(lactide) (PLA), poly(ε-caprolactone) (PCL) and polyhydroxyalkanoates (PHA), the favorable aspects of biodegradable polyurethane are its mechanical, physicochemical and biological properties can be tailored by varying the three blocks in polyurethane backbone, diisocyanate, diol and chain extender. In this thesis, we focus on the development of different functional biodegradable polyurethanes to study their structure-property relationships and satisfy different requirements in cardiac tissue repair and regeneration. We firstly designed a series of degradation-controllable polyurethanes containing various amounts of disulfide bonds in the backbones (PU-SS), which v made the polyurethane selectively sensitive to antioxidants. Glutathione (GSH) is a natural antioxidant existing in intracellular and extracellular fluids as well as in extracellular matrices (ECM). The in vivo degradation of the PU-SS can be initiated by the GSH existing in the cardiac ECM and accelerated via intramuscular injection of GSH to induce the fast degradation of the implant on demand. The PU-SS polymer can be processed into films and electrospun fibrous scaffolds. Synthesized materials exhibited robust mechanical properties and high elasticity. Accelerated degradation of the materials was observed in the presence of GSH, and the rate of such degradation depends on the amount of disulfide present in the polymer backbone. The polymers and their degradation products exhibited no apparent cell toxicity while the electrospun scaffolds supported fibroblast growth in vitro. The in vivo subcutaneous implantation model showed that the polymers prompt minimal inflammatory responses, and as anticipated, the polymer with the higher disulfide bond amount had faster degradation in vivo. Secondly, we developed series of electroactive biodegradable elastomeric polyurethanes as potential cardiac patch because electromechanical coupling of myocytes is crucial for their synchronous response to electrical pacing signals and conductive material can improve the ability of cardiac patch to contract effectively as a unit. First of all, a biodegradable conductive polyurethane (CPU) was synthesized from polycaprolactone diol (PCL), hexadiisocyanate (HDI), and aniline trimer and subsequently doped with (1S)-(+)-10-camphorsulfonic acid (CSA). The electrical conductivity of the CPU films, as enhanced with increasing amounts of CSA, ranged 4.2±0.5 × 10-8 to 7.3±1.5 × 10-5 S/cm in a wet state. The initial moduli of CPU films vi increased from 7±1 MPa to 35±11 MPa with increased CSA amount. The doped CPU film could maintain 87% of initial conductivity after 150 hours charge under physiological environment. After 7 days of enzymatic degradation, the conductivity of all CSA-doped CPU films had decreased to that of the undoped CPU film. Furthermore, most CPU films showed high cell viability except for the group with highest CSA amount. Hence, we found the addition of dopant, CSA, can increase the polymer stiffness and deteriorated its electrical stability and biocompatibility due to the dopant leaching-out. To address this issue, we improved our design by introducing the dopant molecule into the polyurethane backbone via covalent bonding to form a biodegradable, dopant-free conductive polyurethane (DCPU) without adding extra dopant, which possessed improved mechanical properties, electrical conductivity and conductive stability, compared with the CPU polymer. Specifically, a biodegradable PCL, conductive aniline trimer, and dopant dimethylolpropionic acid (DMPA) were linked into DCPU polymer chain through HDI. The electrical conductivities of DCPU s increased with increasing DMPA amounts, ranging from 4.4 ± 0.4 × 10–7 to 4.7 ± 0.8 × 10–3 S/cm in wet state, which were higher than those of CPU polymers doped with different amounts of CSA. The initial moduli of CPU films ranged from 3.0±0.6 MPa to 5.2±1.1 MPa with decreasing DMPA content, which were lower than those of CPU films. The DCPU film showed excellent electrical stability (264% of initial conductivity after 150h charge) under a physiological condition. Furthermore, mouse 3T3 fibroblasts proliferated on these films exhibiting good cytocompatibility. DCPU can also be processed into a porous scaffold using salt leaching. In vivo mouse subcutaneous model exhibited good tissue compatibility with vii extensive cell infiltration over 4 weeks of the DCPU scaffold compared to PCL porous scaffold. Subsequently, because tissue engineered cardiac patch is also required to have mechanical and bioactive properties mimicking the native myocardium, we firstly developed a family of biodegradable elastomeric polyurethanes (PU) with low initial moduli to mechanically mimic the native myocardium. Specifically, random copolymers poly(δ-valerolactone-co-ε-caprolactone) (PVCL) and hydrophilic poly(ethylene glycol) (PEG) were combined into a triblock copolymer, PVCL−PEG−PVCL, and was used as a soft segment in the polyurethane backbone. The triblock copolymers were varied in chemical components, molecular weights, and hydrophilicities. The mechanical properties of polyurethanes in dry and wet states can be tuned by altering the molecular weights and hydrophilicities of the soft segments. Increasing the length of either PVCL or PEG in the soft segments reduced initial moduli of the polyurethane films and scaffolds in dry and wet states. The polymer films are found to have good cell compatibility and to support fibroblast growth in vitro. Selected polyurethanes were then processed into anisotropic porous scaffolds by a thermally induced phase-separation technique and their mechanical properties can be tailored by altering the soft segment molecular weight in the polyurethane backbone, and subsequently by varying the polymer concertation parameters during the scaffold fabrication process. The uniaxial mechanical properties, suture retention strength, ball-burst strength and biaxial mechanical properties of the polyurethane anisotropic porous scaffolds were optimized to mechanically match the native myocardium. The optimal PU-PEG1K-PVCL6K-10% scaffold had ball burst strength viii (20.7 ± 1.5 N) comparable to that of native porcine myocardium (20.4 ± 6.0 N) and showed anisotropic mechanical behavior close to the biaxial behavior of the native porcine myocardium. Furthermore, the optimized