Date of Award

Spring 5-2020

Document Type

Thesis

College

College of Arts and Sciences

Degree Program

Biology

Degree Name

Master of Science

Thesis Advisor

Dr. Matthew Stern

Committee Member

Dr. Kristi Westover

Committee Member

Dr. Laura Glasscock

Abstract

Cardiovascular disease exerts a consistent force of morbidity and mortality on the global population. This pressure has created a demand for reliable replacement vascular tissues. Current solutions include synthetic grafts and biological transplant options that are associated with a multitude of complications. Therefore, the continued innovation of vascular tissue replacement solutions is warranted. Decellularized vascular tissues have been previously utilized as scaffolds upon which recellularized tissue engineered vascular solutions are constructed. However the interplay between decellularization agent concentration and effective recellularization has not been well documented. Herein we investigate this relationship through a detail oriented research model that both qualifies characteristic differences in decellularized tissues prepared in heterogeneous detergent concentrations and quantifies endothelial cell responses to those variations. In brief, seeding scaffolds were created from porcine internal thoracic arteries through detergent based decellularized in five distinct decellularization detergent conditions. These scaffolds were then seeded with endothelial cells using 3D printed seeding windows of our own design. The results of these seeding trials were then evaluated metabolically and through fluorescent microscopy. Additionally, a subset of scaffolds were exempted from seeding trials and examined using scanning electron microscopy to determine detergent concentration effect on scaffold ultrastructure. Results from the metabolic analysis of seeded scaffolds reveals a positive correlation between decellularization detergent concentration and metabolic activity. Additionally, cellular density and decellularization detergent concentration are also positively correlated resulting in significant differences in cell density between detergent concentration groups. However, detergent concentration did not affect the general cellularity of scaffold nor did it cause consistent variation in scaffold ultrastructure.

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