DOCSLIB.ORG

PLGA) Nanoparticles Across the Nasal Mucosa

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
PLGA) Nanoparticles Across the Nasal Mucosa - In vitro assessment of the transport of Poly D, L Lactic-Co-Glycolic Acid (PLGA) nanoparticles across the nasal mucosa Albarki, Mohammed Abdulhussein Handooz https://iro.uiowa.edu/discovery/delivery/01IOWA_INST:ResearchRepository/12730608840002771?l#13730783320002771 Albarki, M. A. H. (2016). In vitro assessment of the transport of Poly D, L Lactic-Co-Glycolic Acid (PLGA) nanoparticles across the nasal mucosa [University of Iowa]. https://doi.org/10.17077/etd.cl2e1klm https://iro.uiowa.edu Copyright 2016 Mohammed Abdulhussein Handooz Albarki Downloaded on 2021/09/27 13:25:19 -0500 - IN VITRO ASSESSMENT OF THE TRANSPORT OF POLY D, L LACTIC-CO- GLYCOLIC ACID (PLGA) NANOPARTICLES ACROSS THE NASAL MUCOSA by Mohammed Abdulhussein Handooz Albarki A thesis submitted in partial fulfillment of the requirements for the Master of Science degree in Pharmacy in the Graduate College of The University of Iowa August 2016 Thesis Supervisor: Professor Maureen D. Donovan Graduate College The University of Iowa Iowa City, Iowa CERTIFICATE OF APPROVAL ____________________________ MASTER'S THESIS _________________ This is to certify that the Master's thesis of Mohammed Abdulhussein Handooz Albarki has been approved by the Examining Committee for the thesis requirement for the Master of Science degree in Pharmacy at the August 2016 graduation. Thesis Committee: __________________________________________ Maureen D. Donovan, Thesis Supervisor __________________________________________ Aliasger K. Salem __________________________________________ Lewis L. Stevens To my parents for their continuous guidance throughout my life and career ii ACKNOWLEDGEMENTS I would like to express my deepest appreciation to my advisor, Professor Dr. Maureen Donovan. This thesis would not have been possible unless her thoughtful support and guidance. Thank you for your expert advices and for patiently teaching me everything I need to know along the way in my graduate education. I would like to thank Professor Dr. Aliasger Salem for his generous gift of PLGA polymer used in this study and for allowing me to use instruments in his laboratory. I would like to express my special thanks for Professor Dr. Lewis Stevens for serving in my thesis committee and for his time in reviewing this work. I would like to thank Kareem Ebeid from Dr. Salem laboratory for his suggestions in nanoparticle preparation and for performing SEM images of my particles. I would also like to thank my labmates in Dr. Donovan Laboratory, Ana Ferreira, Wisam Albakri, Namita Sawant, Laxmi Shanthi, Ammar Alkhafaji, Zainab Bakri, and Saikishore Meruva, for their support in this thesis work. I would also like to thank to Higher Committee for Education Development in Iraq (HCED) for their financial support. Finally, I would like to thank my parents, my brother and sisters for their love and support, my work would not have come so far without them. iii ABSTRACT The nasal mucosa provides a rapid, noninvasive route for drug administration to the systemic circulation and even potentially to the CNS. Nanoparticles made from the biodegradable polymer, PLGA, are of great interest for use in drug delivery systems due to PLGA’s relative safety and ease of surface modification. Nanoparticles may provide improved targeting and transport through the nasal mucosa. However, the optimal nanoparticle sizes and surface properties for intranasal delivery are unknown. In this study, we prepared PLGA nanoparticles within a size range of 50-70 nm containing the lipophilic fluorescent dye, Nile Red, using a surfactant-free nanoprecipitation method. The resulting nanoparticles were evaluated using dynamic light scattering and scanning electron microscopy. Nanoparticle uptake into the nasal mucosa was determined by exposing the tissues to nanoparticle dispersions for 30 or 60 minutes. The in vitro uptake of the nanoparticles by the nasal mucosal tissues revealed that the Nile Red-loaded PLGA nanoparticles were transported across the epithelial layer and accumulated in the sub- mucosal connective tissues. Nanoparticle uptake in the full thickness tissues was time dependent where 2% of the total loads of nanoparticles exposed to the tissues were measured in the mucosal tissue after 30 minutes and 4% were present in the tissues after 60 minutes. The rapid and measurable transfer of PLGA nanoparticles into the nasal mucosal tissues indicate that they may be an efficient delivery vehicle for drugs with either local or systemic activities. iv PUBLIC ABSTRACT Intranasal drug administration provides a rapid, noninvasive route for drug administration directly to the blood and even potentially to the brain. Nanoparticles are particles with diameter < 100 nm that can be used as drug carriers due to their abilities to bypass various biological barriers due to their small sizes. Nanoparticles used in drug delivery are usually made from biocompatible, biodegradable polymers such as poly D, L lactic- co-glycolic acid (PLGA) which can deliver drug without causing long term damage or toxicity. PLGA nanoparticles are of great interest for use as drug carriers due to their relative safety and ease of surface modification. Nanoparticles may provide improved targeting and transport through the nasal mucosa, however, their optimal size and surface properties for effective intranasal delivery are unknown. In this study, PLGA nanoparticles within a size range of 50-70 nm were prepared using a surfactant-free nanoprecipitation method. The resulting nanoparticles were characterized with dynamic light scattering and scanning electron microscopy for size and shape and by Nano Zeta Sizer for surface charge. Nanoparticle uptake into the nasal mucosa was determined by exposing the tissues to nanoparticle dispersions for 30 or 60 minutes. The in vitro uptake of the nanoparticles by the nasal mucosal tissues revealed that Nile Red-loaded PLGA nanoparticles were transported across the epithelial layer and accumulated in the sub-mucosal connective tissues. The rapid and quantitative transfer of PLGA nanoparticles into the nasal mucosal tissues indicated that these ~50 nm particles may be an efficient delivery vehicle for drugs for either local or potentially systemic activities. v TABLE OF CONTENTS LIST OF TABLES ......................................................................................................................... vii LIST OF FIGURES ...................................................................................................................... viii CHAPTER 1: INTRODUCTION .................................................................................................... 1 1.1 Nanoparticles as Drug Carrier............................................................................................ 1 1.2 Biodegradable Polymers in Pharmaceutical Drug Delivery System .................................. 2 1.3 Intranasal Drug Delivery .................................................................................................... 4 1.4 Nasal Anatomy and Histology ........................................................................................... 5 1.5 Nanoparticle Internalization in the Nasal Mucosa ............................................................. 9 1.6 Effect of Nanoparticle Size on the Uptake in the Nasal Mucosa ..................................... 12 CHAPTER 2: OBJECTIVES ......................................................................................................... 14 CHAPTER 3: MATERIALS AND METHODS ........................................................................... 15 3.1 Materials .......................................................................................................................... 15 3.2 Fabrication of PLGA Nanoparticles Using Nanoprecipitation Method ........................... 16 3.3 Preparation of Nanoparticle Using Surfactant-free Nanoprecipitation Method ............... 19 3.4 Preparation of Nanoparticle Dispersion for Tissue Uptake ............................................. 23 3.5 Nile Red Fluorescent Dye ................................................................................................ 28 3.6 Nanoparticle Uptake Studies ............................................................................................ 28 3.7 Calculation of the Number of PLGA Nanoparticles ........................................................ 32 3.8 Quantification of Nile Red Mass Per Single Nanoparticle .............................................. 33 3.9 Data Analysis ................................................................................................................... 34 CHAPTER 4: RESULTS AND DISCUSSION ............................................................................. 35 4.1 Nanoparticle Preparation Using Nanoprecipitation Method ............................................ 35 4.2 Nanoparticle Preparation with the Surfactant-Free Nanoprecipitation Method ............... 39 4.3 Lucifer Yellow VS Transport .......................................................................................... 50 4.4 Quantification of Nanoparticle Uptake ............................................................................ 52 4.5 Conclusion ....................................................................................................................... 58 APPENDIX A: STANDARD CURVES ....................................................................................... 60 APPENDIX B: CALCULATION OF THE PLGA POLYMER DENSITY ................................
Load more

Recommended publications
  • WO 2017/223239 Al 28 December 2017 (28.12.2017) W !P O PCT
    (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization I International Bureau (10) International Publication Number (43) International Publication Date WO 2017/223239 Al 28 December 2017 (28.12.2017) W !P O PCT (51) International Patent Classification: Published: A61K 31/445 (2006.01) C07D 471/04 (2006.01) — with international search report (Art. 21(3)) A61K 31/437 {2006.01) — before the expiration of the time limit for amending the (21) International Application Number: claims and to be republished in the event of receipt of PCT/US20 17/038609 amendments (Rule 48.2(h)) (22) International Filing Date: 2 1 June 2017 (21 .06.2017) (25) Filing Language: English (26) Publication Language: English (30) Priority Data: 62/352,820 2 1 June 2016 (21 .06.2016) 62/456,526 08 February 2017 (08.02.2017) (71) Applicant: X4 PHARMACEUTICALS, INC. [US/US]; 955 Massachusetts Avenue; 4th Floor, Cambridge, Massa chusetts 02139 (US). (72) Inventors: BOURQUE, Elyse Marie Josee; 3115 Racine Street, Unit 214, Bellingham, Washington 98226 (US). SK- ERLJ, Renato; 12 Crocker Circle, West Newton, Massa chusetts 02465 (US). (74) Agent: REID, Andrea L., C. et al; One International Place, 40th Floor, 100 Oliver Street, Boston, Massachusetts 021 10-2605 (US). (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW.
    [Show full text]
  • Cascade Blue and Lucifer Yellow Probes
    Product Information Revised: 24–July–2003 Cascade Blue ® and Lucifer Yellow Probes Fixable Polar Tracers salt (L-1177, solu bil i ty ~1%) or the am mo ni um salt (L-682, sol- u bil i ty ~6%) may be pre ferred in ap pli ca tions where lith i um ions Molecular Probes prepares a wide variety of highly water- in ter fere with bi o log i cal function. sol u ble dyes and other detectable probes that can be used as cell Although its low absorbance at 488 nm (⑀ ~700 cm-1M-1) trac ers. Polar tracers can also be incorporated into lip o somes makes it inefficiently excited with the argon-ion laser, lucifer to gen er ate polymeric fluorescent tracers or antibody-la bel ing yellow CH has been used as a neuronal tracer in confocal la ser reagents .1-5 scan ning micros copy studies.19-21 For electron mi crosco py stud ies, In most cases, these water-soluble probes are too polar to lucifer yel low can be used to photo convert diaminobenzidine passive ly diffuse through cell membranes. Consequently, special (DAB) into an insolub le, electron-dense re ac tion product.22-24 meth ods for loading the dyes must be employed, in clud ing mi cro- Al ter na tive ly, anti–lucifer yellow dye an ti bod ies may be used inje ction, pino cytosis or techniques that tem po rari ly per me abilize with en zyme-mediated immuno histochemical method s to de vel op the cellʼs membrane.
    [Show full text]
  • Fluorescent Cellular Stains
    Fluorescent Cellular Stains Membrane and Membrane Potential Dyes CellBrite™ Cytoplasmic Membrane Stains and Other Lipophilic Carbocyanine Dyes ... p. 2 Phospholipid Membrane Dyes ... p. 3 Membrane Potential Dyes ... p. 3 Synaptic Vesicle Dyes ... p. 4 Cytosolic Tracers ... p. 5 Organelle Stains MitoView™ Mitochondrial Dyes ... p. 6 LysoView™ Lysosome Stains ... p. 7 Cytoskeleton Probes ... p. 8 Nuclear Stains ... p. 9 Apoptosis and Viability Stains ... p. 10 Fluorescent Lectins, Toxins, and Other Conjugates ... p. 11 www.biotium.com • 1 Fluorescent Membrane and Membrane Potential Dyes CellBrite™ Cytoplasmic Membrane Dyes Lipophilic carbocyanine dyes are widely used for labeling neurons in tissues by retrograde labeling, and to label membranes in a wide variety of cell types. The dyes are weakly fluorescent in aqueous phase, but become highly fluorescent in lipid bilayers. Staining is highly stable with low toxicity and very little dye transfer in between cells, making the dyes suitable for long-term cell labeling and tracking studies. Cell populations can be labeled with different fluorescent colors for identification after mixing. Double labeling can identify cells that have fused or formed stable clusters. Unlike PKH dyes, CellBrite™ dyes do not require a complicated hypoosmotic labeling protocol. They are ready-to-use dye delivery solutions that can be added directly to normal culture media to label suspended or adherent cells in culture. We offer a selection of dyes with fluorescence ranging from blue to near-infrared. The CellBrite™ Blue Cytoplasmic Membrane Labeling Kit features DiB, a unique blue carbocyanine dye, and a cell loading agent. CellBrite™ Green (NeuroDiO), CellBrite™ Orange (DiI), CellBrite™ Red (DiD), and CellBrite™ NIR dyes are supplied as ready-to-add dye solutions.
    [Show full text]
  • Handbook of Biological Dyes and Stains Synthesis and Industrial Applications
    HANDBOOK OF BIOLOGICAL DYES AND STAINS SYNTHESIS AND INDUSTRIAL APPLICATIONS R. W. SABNIS Pfizer Inc. Madison, NJ HANDBOOK OF BIOLOGICAL DYES AND STAINS HANDBOOK OF BIOLOGICAL DYES AND STAINS SYNTHESIS AND INDUSTRIAL APPLICATIONS R. W. SABNIS Pfizer Inc. Madison, NJ Copyright Ó 2010 by John Wiley & Sons, Inc. All rights reserved. Published by John Wiley & Sons, Inc., Hoboken, New Jersey Published simultaneously in Canada No part of this publication may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, recording, scanning, or otherwise, exckpt as permitted under Section 107 or 108 of the 1976 United States Copyright Act, without either the prior written permission of the Publisher, or authorization though payment of the appropriate per-copy fee to the Copyright Clearance Center, Inc., 222 Rosewood Drive, Danvers, MA 01923, (978) 750-8400, fax (978) 750-4470, or on the web at www.copyright.com. Requests to the Publisher for permission should be addressed to the Permissions Department, John Wiley & Sons, Inc., 111 kver Street, Hoboken, NJ 07030, (201) 748-601 1, fax (201) 748-6008, or online at http://www.wiley.com/go/permission. Limit of Liability/Disclaimer of Warranty: While the publisher and author have used their best efforts in preparing this book, they make no representations or warranties with respect to the accuracy or completeness of the contents of this book and specifically disclaim any implied warranties of merchantability or fitness for a particular purpose. No warranty may be created or extended by sales representatives or written sales materials.
    [Show full text]
  • Lucifer Yellow
    FluoProbes® FT-15437A Lucifer Yellow Products Description Name : Lucifer yellow CH lithium salt Benz[de]isoquinoline-5,8-disulfonic acid, 6-amino-2-[(hydrazinocarbonyl)amino]-2, 3-dihydro-1,3-dioxo-, dilithium salt; 6-Amino-2,3-dihydro-1,3-dioxo-2-hydrazinocarbonylamino-1H-benz[d,e]isoquinoline-5,8- disulfonic acid dilithium salt Catalog Number : FP-15437A, 25 mg Structure : C13H9Li2N5O9S2 Molecular Weight : 457.24 Solubility: In DMF, DMS and water Absorption / Emission : λexc\λem = 428 nm/536 nm EC (M-1 cm-1) : 12 000 Name : Lucifer yellow CH potassium salt 1H-Benz[de]isoquinoline-5,8-disulfonic acid, 6-amino-2-(hydrazinocarbonyl)amino]-2, 3-dihydro-1,3- dioxo-, dipotassium salt Catalog Number : FP-52489A, 25 mg Structure : C13H9K2N5O9S2 Molecular Weight : 521.6 Solubility: In water Absorption / Emission : λexc\λem = 427 nm/535 nm EC (M-1 cm-1) : 12 000 Also available: Lucifer yellow ethylenediamine FP-53451A, 25 mg 1H-Benz[de]isoquinoline-5,8-disulfonic acid, 6-amino-2-(2-aminoethyl)-2,3-dihydro-1,3-dioxo-, dipotassium salt ; N-(2-Aminoethyl)-4-amino-3,6-disulfo-1,8-naphthalimide, dipotassium salt MW: 491.6 Soluble in DMSO, DMF, water λexc\λem = 426 nm/ 531 nm References: Polson NA, Hayes MA. Anal Chem 72, 1088-1092 (2000) [Water soluble polar tracer] Lucifer Yellow anhydride dipotassium salt FP-BT8930 [4-Amino-3,6-disulfo-1,8-naphthalic anhydride dipotassium salt, 1H,3H-Naphtho[1,8-cd]pyran-5,8-disulfonic acid, 6-amino-1,3-dioxo-, dipotassium salt ] MW: 449.5 Building block to water soluble polar tracer such as lucifer yellow probes Lucifer yellow cadaverine FP-86413A [N-(2-Aminopentyl)-4-amino-3,6-disulfo-1,8-naphthalene, dipotassium salt ] MW: 533.67 Soluble in DMSO and H2O λexc\λem = 425 nm/ 531 nm Water soluble polar tracer ; (Z) References : 1; Giordano L, Jovin TM, Irie M, Jares-Erijman EA.
    [Show full text]