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Food Protein-Based Core-Shell Nanocarriers for Oral Drug Delivery Applications

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Food Protein-Based Core-Shell Nanocarriers for Oral Drug Delivery Applications South Dakota State University Open PRAIRIE: Open Public Research Access Institutional Repository and Information Exchange Theses and Dissertations 2017 Food Protein-based Core-shell Nanocarriers for Oral Drug Delivery Applications: (Influence of Shell Composition on In vitro and In vivo Functional Performance of Zein Nanocarriers MD Saiful Islam South Dakota State University Follow this and additional works at: https://openprairie.sdstate.edu/etd Part of the Pharmacy and Pharmaceutical Sciences Commons Recommended Citation Islam, MD Saiful, "Food Protein-based Core-shell Nanocarriers for Oral Drug Delivery Applications: (Influence of Shell Composition on In vitro and In vivo Functional Performance of Zein Nanocarriers" (2017). Theses and Dissertations. 2149. https://openprairie.sdstate.edu/etd/2149 This Dissertation - Open Access is brought to you for free and open access by Open PRAIRIE: Open Public Research Access Institutional Repository and Information Exchange. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of Open PRAIRIE: Open Public Research Access Institutional Repository and Information Exchange. For more information, please contact [email protected]. i FOOD PROTEIN-BASED CORE−SHELL NANOCARRIERS FOR ORAL DRUG DELIVERY APPLICATIONS: INFLUENCE OF SHELL COMPOSITION ON IN VITRO AND IN VIVO FUNCTIONAL PERFORMANCE OF ZEIN NANOCARRIERS BY MD SAIFUL ISLAM A dissertation submitted in partial fulfillment of the requirements for the Doctor of Philosophy Major in Pharmaceutical Sciences South Dakota State University 2017 iii THIS DISSERTATION IS DEDICATED TO MY PARENTS iv ACKNOWLEDGMENTS So many peoples have a positive influence throughout my graduate study at South Dakota State University (SDSU). First, I would like to express my heartfelt gratitude to Dr. Omathanu Perumal for his continuous support, motivation, guidance, and supervision during my Ph.D. research work. I also want to express my sincere gratitude and appreciation to advisory committee members including Dr. Xiangming Guan, Dr. Xiuqing Wang and Dr. Alan Davis for their continuous encouragement, direction throughout my Ph.D. degree. I also want to express my gratitude to my previous advisor Dr. Hongwei Zhang for his guidance, support, and supervision for first two years. I would like to thank all my previous and current lab members including Dr. Mohammed Alqahtani, Dr. Kaushalkumar Dave, Dr. Ranjith Averineni, Dr. Manju Saraswathy, Mibin Kuruvilla Joseph and Abdulsalm Alqahtani for their kind help for my rapid progress with the assigned project in the lab. It was my great pleasure to be a member of Pharmacy family at SDSU which reminds me the humble behavior of Dr. Chandradhar Dwivedi. I greatly appreciate Dr. Shafiqur Rahman for his continuous help and motivation during my Ph.D. degree. I would like to thank Dr. Jayarama Gunaje for his motivation and encouragement during my Ph.D. research work. I appreciate the help of Dr. Joshua Reineke by providing enormous useful information and assisting me with bioadhesive experiments. I want to express my gratitude to Dr. Hemchand Tummala for helping me in learning in vivo imaging instrument and supervising fludrocortisone project. I would like to thank Dr. Monzurul Amin Roni, Dr. Muzaffar Abbas and Dr. Sayef Abdullah for their support and helping me with ELISA experiment v I want to thank Dr. James Hoefelmeyer and Dr. Aravind Baride from Department of Chemistry, University of South Dakota, Vermillion for his help with the transmission electron microscopy (TEM) studies. I also want to thank Dr. Jayarama Gunaje for his continuous encouragement and help in Caco-2 cell culture study. I owe my gratitude and utmost respect to my wonderful parents for their unconditional and sincere love, support, and encouragement. I am forever indebted to my dear wife, for her love, patience and exceptional forgiveness that provided me the strength and confidence to complete my dissertation. Lastly, but not least of all, I will always remain grateful to the department of Pharmaceutical Sciences, College of Pharmacy and Allied Health Professions at South Dakota State University, for Ph.D. fellowship and financial support. vi CONTENTS Page no. LIST OF FIGURES----------------------------------------------------------------- xi LIST OF TABLES------------------------------------------------------------------ xv SYMBOLS/ABBREVIATIONS-------------------------------------------------- xviii ABSTRACT-------------------------------------------------------------------------- xx CHAPTER ONE: Introduction------------------------------------------------- 1 1.1.Oral drug delivery--------------------------------------------------------------- 1 1.2. Oral drug absorption----------------------------------------------------------- 1 1.2.1. Gastrointestinal anatomy and physiology------------------------------- 1 1.2.2. Mechanism of oral drug absorption------------------------------------- 4 1.3. Oral drug delivery challenges------------------------------------------------ 12 1.3.1. Physiological barriers to oral drug delivery----------------------------- 12 1.3.2. Influence of physicochemical and biopharmaceutical factors on 19 oral drug delivery----------------------------------------------------------- 1.4. Different approaches for oral drug delivery-------------------------------- 23 1.4.1. Physical or chemical modification of drug molecules----------------- 23 1.4.2. Drug carriers---------------------------------------------------------------- 27 1.4.2.1. Lipid-based delivery systems-------------------------------------------- 29 1.4.2.2. Polymeric nanocarriers--------------------------------------------------- 39 1.5. Natural polymers-------------------------------------------------------------- 46 1.5.1. Polysaccharide-based nanocarriers--------------------------------------- 46 vii 1.5.2. Protein polymers ----------------------------------------------------------- 48 1.5.2.1. Animal proteins----------------------------------------------------------- 50 1.5.2.2. Plant proteins--------------------------------------------------------------- 52 1.6. Scope and objectives--------------------------------------------------------- 57 CHAPTER TWO: Preparation and Study of Structure-Function 59 Relationship of Core-Shell Nanocarriers-------------------------------------- 2.1. Introduction--------------------------------------------------------------------- 60 2.2. Materials and methods--------------------------------------------------------- 65 2.2.1. Materials---------------------------------------------------------------------- 65 2.2.2. Preparation of zein-based core-shell nanocarriers----------------------- 65 2.2.3. In vitro characterization and optimization-------------------------------- 67 2.2.4. Determination of encapsulation and loading efficiency---------------- 67 2.2.5. In vitro release of Nile red (NR)------------------------------------------- 68 2.2.6. Enzymatic degradation of core-shell nanocarriers----------------------- 68 2.2.7. Influence of pH and ionic strength on nanocarriers stability----------- 69 2.2.8. Cell culture-------------------------------------------------------------------- 69 2.2.9. Mechanism of cellular uptake of NR loaded nanocarriers------------- 69 2.2.10. Calcein uptake assay------------------------------------------------------- 70 2.2.11. In vitro transepithelial transport of core-shell nanocarriers----------- 71 2.2.12. Cell uptake studies using confocal laser scanning microscopy------ 71 2.2.13. Ex vivo bioadhesion assay------------------------------------------------- 72 2.2.14. Ex vivo tissue uptake studies using pig jejunum----------------------- 76 viii 2.2.15. Animal studies-------------------------------------------------------------- 75 2.3. Statistical analysis---------------------------------------------------------- 79 2.4. Results and discussion--------------------------------------------------------- 80 2.4.1. Characteristics and stability of core-shell nanocarriers----------------- 80 2.4.2. In vitro release of NR from core-shell nanocarriers--------------------- 96 2.4.3. In vitro transepithelial transport and cell uptake of nanocarriers------ 101 2.4.4. Oral biodistribution of nanocarriers--------------------------------------- 111 2.4.5. Ex vivo bioadhesion assay-------------------------------------------------- 117 2.4.6. Immunogenicity of core-shell nanocarriers------------------------------ 120 2.5. Conclusion---------------------------------------------------------------------- 122 CHAPTER THREE: Oral Delivery of a Model Anti-Retroviral Drug 125 Using Core-Shell Nanocarriers-------------------------------------------------- 3.1. Introduction-------------------------------------------------------------------- 126 3.2. Materials and methods--------------------------------------------------------- 129 3.2.1. Materials---------------------------------------------------------------------- 129 3.2.2. Methods----------------------------------------------------------------------- 129 3.2.2.1. Preparation of nanocarriers----------------------------------------------- 129 3.2.2.2. Characterization of LPV loaded nanocarriers-------------------------- 130 3.2.2.3. Differential Scanning Calorimetry (DSC)------------------------------ 130 3.2.2.4. HPLC analysis of LPV---------------------------------------------------- 130 3.2.2.5. Determination of encapsulation and loading efficiency of LPV---- 131 3.2.2.6. In vitro release of LPV from nanocarriers-----------------------------
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