Master of Science in Biomedical Engineering, Cleveland State University, 2014, Washkewicz College of Engineering

Vascular diseases such as atherosclerosis and aneurysms are characterized by the over-proliferation and migration of aortic SMCs, and degradation of ECM within the vessel wall, leading to compromise in cell-cell and cell-matrix signaling pathways. Recent tissue engineering approaches to regulate SMC over-proliferation and enhance healthy ECM synthesis showed promise, but resulted in low crosslinking efficiency and matrix deposition yields. In this study, the benefits of exogenous nitric oxide cues, delivered from GSNO, to cell proliferation and matrix deposition by adult human aortic SMCs within 3D biomimetic cultures have been explored. The first experiment utilized a microfluidic platform with two adjacent, permeable 3D culture chambers, to enable paracrine signaling between vascular cell cocultures. Healthy HA-SMCs were cultured in these devices within 3D collagen hydrogels, either alone or in the presence of human aortic ECs cocultures, and exogenously supplemented with varying GSNO dosages (0-100 nM) for 21 days. Results showed that EC cocultures stimulated SMC proliferation within GSNO free cultures. However, with increasing GSNO concentration, SMC proliferation decreased in the presence or absence of EC cocultures, while EC proliferation increased. GSNO (100 nM) significantly enhanced the total protein amount synthesized by SMCs, in the presence or absence of EC cocultures, while lower dosages offered marginal benefits. On a per cell basis, multi-fold increases in the synthesis and deposition of elastin, GAGs, hyaluronic acid and LOX were noted at higher GSNO dosages, and coculturing with ECs significantly furthered these trends. The matrix yields of these proteins reached almost 40 - 51 % within selective cocultures receiving GSNO. Similar increases in TIMP-1 and MMP-9 levels were noted within cocultures with increasing GSNO dosages, although MMP-2 levels remained attenuated. These quantitative assay data were strongly supported by immunofluorescence image (open full item for complete abstract)

Committee: Chandra Kothapalli PhD (Committee Chair); Nolan Holland PhD (Committee Member); Moo-Yeal Lee PhD (Committee Member) Subjects: Anatomy and Physiology; Biochemistry; Biomedical Engineering; Biomedical Research; Cellular Biology; Engineering; Health; Health Care; Medicine; Pharmaceuticals