DOI: https://doi.org/10.15368/theses.2017.66
Available at: https://digitalcommons.calpoly.edu/theses/1801
Date of Award
6-2017
Degree Name
MS in Biomedical Engineering
Department/Program
Biomedical and General Engineering
Advisor
Kristen O'Halloran Cardinal
Abstract
The development of tissue engineered blood vessel mimics for the testing of intravascular devices in vitro has been established in the Cal Poly tissue engineering lab. Due to the prevalence of cardiovascular disease in diabetic patients and minimal accessible studies regarding the interactions between diabetes and intravascular devices used to treat vascular disease, there is a need for the development of diabetic models that more accurately represents diabetic processes occurring in the blood vessels, primarily endothelial dysfunction. This thesis aimed to create a diabetic blood vessel mimic by implementing a high glucose environment for culturing human endothelial cells from healthy umbilical veins (HUVECs) and from diabetic coronary arteries (DHCAECs). The characterization of these BVMs was achieved using immunofluorescence, scanning electron microscopy (SEM), and qPCR gene expression analysis. From this study, it was determined that HUVECs and DHCAECs are robust enough to be cultured in a high glucose environment – analogous to hyperglycemia – and these cells exhibited different characteristics when evaluated under microscopy and qPCR gene expression. The immunofluorescence and SEM imaging showed presence of cells within each blood vessel mimic. The qPCR gene expression analysis demonstrated that mRNA expression of endothelial nitric oxide synthase (eNOS), platelet endothelial cell adhesion molecule (PECAM), and receptor for advanced glycation end products (RAGE) differs between HUVECs and DHCAECs, as well as between cells cultured in v normal and elevated glucose concentrations. These differences in gene regulation indicate the potential of the diabetic BVM to more accurately represent the endothelial response to diabetes and to the implementation of intravascular devices in the future. It was determined that culturing DHCAECs in a high glucose cell media for use in blood vessel mimics results in a model that differs considerably from HUVECs grown in normal glucose media. It was also determined that there was a difference between DHCAECs cultured in high glucose media and normal glucose media, as well as HUVECs cultured in high glucose media and normal glucose media. This study aided the development of a diabetic BVM; however, there are still improvements to be made, namely the inclusion of vascular smooth muscle cells in the model and improving the confluency of the BVM.