University of Kentucky UKnowledge Theses and Dissertations--Biomedical Engineering Biomedical Engineering 2014 MUCOADHESIVE FILMS FOR TREATMENT OF LOCAL ORAL DISORDERS: DEVELOPMENT, CHARACTERIZATION AND IN VIVO TESTING Sandeep K. Ramineni University of Kentucky, [email protected] Right click to open a feedback form in a new tab to let us know how this document benefits ou.y Recommended Citation Ramineni, Sandeep K., "MUCOADHESIVE FILMS FOR TREATMENT OF LOCAL ORAL DISORDERS: DEVELOPMENT, CHARACTERIZATION AND IN VIVO TESTING" (2014). Theses and Dissertations-- Biomedical Engineering. 19. https://uknowledge.uky.edu/cbme_etds/19 This Doctoral Dissertation is brought to you for free and open access by the Biomedical Engineering at UKnowledge. It has been accepted for inclusion in Theses and Dissertations--Biomedical Engineering by an authorized administrator of UKnowledge. For more information, please contact [email protected]. STUDENT AGREEMENT: I represent that my thesis or dissertation and abstract are my original work. Proper attribution has been given to all outside sources. I understand that I am solely responsible for obtaining any needed copyright permissions. I have obtained needed written permission statement(s) from the owner(s) of each third-party copyrighted matter to be included in my work, allowing electronic distribution (if such use is not permitted by the fair use doctrine) which will be submitted to UKnowledge as Additional File. I hereby grant to The University of Kentucky and its agents the irrevocable, non-exclusive, and royalty-free license to archive and make accessible my work in whole or in part in all forms of media, now or hereafter known. I agree that the document mentioned above may be made available immediately for worldwide access unless an embargo applies. I retain all other ownership rights to the copyright of my work. I also retain the right to use in future works (such as articles or books) all or part of my work. I understand that I am free to register the copyright to my work. REVIEW, APPROVAL AND ACCEPTANCE The document mentioned above has been reviewed and accepted by the student’s advisor, on behalf of the advisory committee, and by the Director of Graduate Studies (DGS), on behalf of the program; we verify that this is the final, approved version of the student’s thesis including all changes required by the advisory committee. The undersigned agree to abide by the statements above. Sandeep K. Ramineni, Student Dr. David Puleo, Major Professor Dr. Abhijit R Patwardhan, Director of Graduate Studies MUCOADHESIVE FILMS FOR TREATMENT OF LOCAL ORAL DISORDERS: DEVELOPMENT, CHARACTERIZATION AND IN VIVO TESTING _________________________________________________ DISSERTATION __________________________________________________ A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the College of Engineering at the University of Kentucky By Sandeep Krishna Ramineni Lexington, Kentucky Director: Dr. David A. Puleo, Professor Department of Biomedical Engineering Lexington, Kentucky 2013 Copyright © Sandeep Krishna Ramineni 2013 ABSTRACT OF THIS DISSERTATION MUCOADHESIVE FILMS FOR TREATMENT OF LOCAL ORAL DISORDERS: DEVELOPMENT, CHARACTERIZATION AND IN VIVO TESTING Mucoadhesive drug delivery systems which are being used from 1980’s to avoid first pass metabolism of drugs, commercially exist for only systemic drug delivery with fast erosion times (15-60 min), that may not be appropriate for local oral disorders. The goal of this research was to develop and characterize mucoadhesive films with flexibility of carrying different drugs and proteins and provide sustained release for local treatment of oral disorders. Mucoadhesive films composed of polyvinylpyrrolidone (PVP) and carboxymethlycellulose (CMC) were formulated with imiquimod, an immune response modifier. Problems such as solubilization of imiquimod to increase drug loading, uniformity in films and total amount of drug released into supernatants were addressed by use of acetate buffer after investigating multiple methods. Subsequently, other relevant properties of mucoadhesive systems, such as adhesion (shear, pull-off), tensile properties, swelling profiles, transport kinetics, and subsequent changes in release profiles as a function of film composition were characterized. The potential of the system for local retention of imiquimod, determined in oral mucosa of hamsters showed time dependent decrease in imiquimod amount through 12 hours, with no traces of drug in blood. Further testing in humans revealed that the residence time of the mucoadhesive films depended on the application site, increasing in the order of tongue < cheek < gingiva. In parallel, mucoadhesive films loaded with epidermal growth factor (EGF) were developed to promote treatment of oral mucosal wounds. Bioactivity was tested in vitro on buccal tissues by creating a wound followed by application of films. Although EGF-loaded films did not accelerate wound healing, but rather elicited a hyperparakeratotic response. In vitro buccal tissues may not be appropriate for testing the effects of EGF in wound healing without incorporation of other biochemical factors. Overall, a mucoadhesive system capable of delivering bioactive small molecules and proteins in sustained manner was developed in this work. A thorough understanding of the system properties was achieved to further tune for future applications. In vitro studies and in vivo studies in hamsters and humans clearly showed the potential and usefulness of the system to translate in to clinic for treatment of oral precancerous lesions. KEYWORDS: Mucoadhesive films, imiquimod, oral dysplasia, mucosal wound healing, local treatment. Sandeep Krishna Ramineni Student’s Signature 06-03-2014 Date MUCOADHESIVE FILMS FOR TREATMENT OF LOCAL ORAL DISORDERS: DEVELOPMENT, CHARACTERIZATION AND IN VIVO TESTING By Sandeep Krishna Ramineni Dr. David Puleo Director of Dissertation Dr. Abhijit R Patwardhan Director of Graduate Studies 06-03-2014 Date Acknowledgements It gives me great pleasure in acknowledging the support and help of many people for the successful completion of this dissertation. I would like to express the deepest appreciation to my committee chair, advisor, Professor David Puleo for accepting me in to this program and offer to work in his lab. I am grateful for his patience during my initial stages of work and further guidance towards my professional development. I would like to thank for his continued support through advices and scholarships, which provided me peace of mind in performing the work. Without his guidance and persistent help this dissertation would not have been possible. I consider it an honor to work with renowned oral and maxillofacial surgeon Dr. Larry Cunningham and am thankful for his suggestions and his practical help in this work. I would like to take this opportunity to thank Dr. Thomas Dziubla for his key suggestions and allowed to use his lab resources when needed. I would also like to thank all my other committee members Dr. Hainsworth Shin, Dr. Babak Bazrgari for their contributions to this work. I am grateful for my funding sources National Institute of Health (NIH) and National Aeronautics and Space Administration Experimental Program to Stimulate Competitive Research (NASA EPSCoR), without which this study would not have been possible. I am obliged to all my lab members for their ideas during several intellectual discussions which have helped in improving the research productivity. I would like to dedicate this dissertation to my parents who have given me the opportunity of an education from the best institutions and support throughout my life. iii TABLE OF CONTENTS Acknowledgements ............................................................................................................ iii LIST OF TABLES ............................................................................................................. ix LIST OF FIGURES ............................................................................................................ x Chapter 1 Introduction ........................................................................................................ 1 Chapter 2 Background ........................................................................................................ 3 2.1 Biology of Oral Mucosa ............................................................................................ 3 2.1.1 Structure.............................................................................................................. 3 2.1.2 Mucus ................................................................................................................. 4 2.2 Drug Delivery to Oral Mucosa .................................................................................. 4 2.3 Mucoadhesive Drug Delivery Systems ..................................................................... 6 2.3.1 Mechanism of action .......................................................................................... 6 2.3.2 Factors affecting mucoadhesion ......................................................................... 7 2.4 Mucoadhesive Polymers ........................................................................................... 7 2.4.1 Natural polymers ................................................................................................ 8 2.4.2 Synthetic polymers ............................................................................................