Author ORCID Identifier
Date Available
7-13-2021
Year of Publication
2021
Degree Name
Doctor of Philosophy (PhD)
Document Type
Doctoral Dissertation
College
Engineering
Department/School/Program
Chemical and Materials Engineering
First Advisor
Dr. Bradley J. Berron
Abstract
Cardiovascular disease remains the number one threat to American lives. During an acute myocardial infarction (AMI), blood flow is blocked and results in the formation of scar tissue. As the body’s immune system responds, inflammatory signaling causes an increase in both scar tissue size and the patient’s risk for further chronic heart failure. In order to reduce the risk of continued heart disease inflammatory signaling must be reduced. Stem cell therapies have the ability to alter the immune system’s pro-inflammatory signal. However, stem cell retention is limited due to blood flow shear. Gelatin methacrylate (GelMA) based coatings have been shown to increase the retention of these therapeutic cells inside the infarcted myocardium. This dissertation evaluates the GelMA coating process and aims to optimize this therapy for future trials.
Biotin-streptavidin (SA) affinity is the foundation for our GelMA coating process. GelMA coatings are grown from eosin bound to the cell surface through a biotin-SA complex. In this work, we demonstrate the high binding affinity through a cardiac flow model. We show that biotinylated cardiac cells can be immobilized onto a SA functionalized glass substrate and withstand large shear. While biotin-SA affinity allows for glass immobilization, we have yet to study what drives the adhesion of our biotin-SA grown GelMA coatings in vivo. We examine two different binding sites for GelMA coated cells: the extracellular matrix (ECM) or resident cells that make up the myocardium. GelMA is created from the degradation of collagen, however, our studies show collagen-binding integrins on the surface of cells do not promote adhesion of GelMA. Through decellularization of heart ECM, we were able to show GelMA-ECM is a strong binding mode that is driven by collagen-gelatin binding. As our coatings drive adhesion through collagen-collagen binding, I hypothesize that increasing the amount of GelMA on the cell surface will result in increased retention in vivo. In our previous work, cells were biotinylated through covalent attachment to free cell surface amines. In an effort to increase the density of biotin on a cell surface, we utilize a lipid insertion approach. The use of cholesterol anchors increased streptavidin density on the surface of cells further driving polymerization and allowing for an increased fraction of cells coated with gelatin (83%) when compared to covalent methods (52%). Altogether, through the use of cholesterol anchors and our in vivo understanding of coating specificity, we have created a path towards improving the retention of MSC in vivo for post-MI therapy.
Digital Object Identifier (DOI)
https://doi.org/10.13023/etd.2021.247
Funding Information
Kara Davis Funding:
National Institutes of Health National Center for Advancing Translational Sciences, TL1TR001997, Year: 2019-2021
Berron Lab Funding:
National Heart, Lung and Blood Institute (NHLBI), R01 HL127682, Year: 2015-2019
National Science Foundation, CBET-1351531, Year: 2018-2021
American Heart Association, #18IPA34170059, Year: 2019-2020
Recommended Citation
Davis, Kara Amelle, "Optimization of Gelatin-Based Cellular Coating of MSC for Myocardial Infarction Therapy" (2021). Theses and Dissertations--Chemical and Materials Engineering. 132.
https://uknowledge.uky.edu/cme_etds/132
Included in
Biochemical and Biomolecular Engineering Commons, Molecular, Cellular, and Tissue Engineering Commons
