Access Type

Open Access Dissertation

Date of Award

January 2020

Degree Type

Dissertation

Degree Name

Ph.D.

Department

Biomedical Engineering

First Advisor

Mai T. Lam

Second Advisor

Jian-Ping Jin

Abstract

Creation of an in vitro atherosclerotic disease model using the novel Ring Stacking Method. Singular self-assembling tissue rings made up smooth muscle cells and fibrin hydrogel are stacked on one another to create a tissue engineered vessel. These biologically engineered blood vessels are then seeded with endothelial cells via combined static rotational and dynamic bioreactor in order to create a functional intima layer. Early stage atherosclerosis was induced via the addition of oxidized low-density lipoproteins (ox-LDL) to the fibrin hydrogel that creates the media layer of the engineered vessel. After the creation of the intima layer the engineered vessel was then statically seeded with THP-1 monocytes differentiated into M1 macrophages via phorbol 12-myristate 13-acetate (PMA), lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma) for 24 hours over the course of 3 days. On the second and third day calci-protein particles (CPPs) along with fibroblast-derived growth factor (FGF) and platelet-derived growth factor (PDGF) were added to the THP-1 differentiation suspension. Following static macrophage seeding the engineered disease vessel was frozen, sectioned and stained via an Oil Red O stain to visualize the macrophage migration and uptake of the ox-LDLs in the hydrogel. Late stage atherosclerotic calcification was characterized via the incorporation of CPPs into engineered vessels. These vessels were mechanically tested, and the data was incorporated into a system coupled computer fluid dynamic (CFD) simulation via ANSYS 19. Engineered disease media (EDM) rings pre-seeded with smooth muscle cells, ox-LDLs, undifferentiated THP-1 cells and CPPs were mechanically and histologically characterized.

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