Please use this identifier to cite or link to this item:
http://hdl.handle.net/11375/28009
Title: | DESIGNING CELL- AND PROTEIN-BASED IN VITRO ASSAYS AS MODELS FOR FIBROTIC RESPONSES TO IMPLANTED HYDROGEL CAPSULES |
Other Titles: | ASSAY DESIGN FOR IMMUNOLOGICAL RESPONSES ON POLYMER CAPSULES |
Authors: | Raez-Villanueva, Sergio |
Advisor: | Holloway, Alison |
Department: | Medical Sciences |
Keywords: | Foreign Body Responses;Fibrosis;Hydrogel Capsules;Cell Encapsulation;Biocompatibility;Diabetes Mellitus;Cell Therapy;Polymer Coatings |
Publication Date: | Nov-2022 |
Abstract: | It is projected that, by 2030, 8% of all adults in the world will have diabetes mellitus and treatment will account for 10% of the total healthcare budget in many countries. Polymeric biomaterial research has led to the design of robust polymer hydrogel capsules to develop curative cell-based therapies for chronic disorders such as diabetes mellitus. Encapsulation of insulin-producing beta cells within synthetic, semi-permeable polymer hydrogels can avoid host immune rejection including fibrotic responses, and thus holds the promise of a long-term curative treatment of this disease. There is a paucity of literature regarding methods available for standardized in vitro screening of synthetic polymer hydrogel capsules to predict host responses in vivo. Thus, the focus of this thesis was to design in vitro assays able to screen for subsequent in vivo fibrotic responses. Two dimensional (‘2D’) (cell attachment to thin film hydrogel coatings) and three dimensional (‘3D’) (cell attachment and protein adsorption to hydrogel capsules) in vitro experiments were designed and tested in an iterative process to assess fibrotic responses to a diverse group of polymer hydrogels. Cell attachment assays included fibroblast (NIH 3T3) and macrophage (RAW 264.7) cell lines, and protein adsorption assays included proteins used to model fibrosis including fibrinogen and lysozyme. For some formulations, in vitro assays were compared with in vivo data on pericapsular cellular overgrowth (PCO) after being implanted into mice. A binomial logistic regression model was designed and validated to assess whether the ‘3D’ in vitro assays correlated with in vivo PCO responses. It was found that the RAW 264.7 cell attachment assay was significantly correlated with PCO outcomes in vivo, demonstrating for the first time a simple, cost-effective, and rapid in vitro cell-based approach to screen and select capsules with lower fibrotic potential to be further tested in animals. |
Description: | For a lay summary of the thesis presented in a 1-minute video format, visit the following link: https://www.youtube.com/watch?v=VhLzt_tEz-s |
URI: | http://hdl.handle.net/11375/28009 |
Appears in Collections: | Open Access Dissertations and Theses |
Files in This Item:
File | Description | Size | Format | |
---|---|---|---|---|
Raez-Villanueva_Sergio_finalsubmission2022August_MSc.pdf | Final MSc thesis by Sergio Raez-Villanueva | 3.11 MB | Adobe PDF | View/Open |
Items in MacSphere are protected by copyright, with all rights reserved, unless otherwise indicated.