Combined International Symposium for Applied Cardiovascular Biology and Vascular Tissue Engineering, June 19-21, Zurich, Switzerland
Razvan D. Macarie, Sergiu CecoltanLetitia Ciortan, Agneta Simionescu, Elena Butoi, Ileana Manduteanu
Institute of Cellular Biology and Pathology “Nicolae Simionescu”, Bucharest, Romania
Diabetes accelerates calcific aortic valve disease (CAVD) and is predictive of poor prognosis in valve disease and faster degeneration of implanted bio-prosthetic aortic valves. CAVD is a complex pathology that involves several biological processes from lipid accumulation, inflammation, remodeling, osteogenesis and finally gross calcification and compromised valve function. The molecular mechanisms of extracellular matrix (ECM) remodeling in CAVD still need to be uncovered. The aim of this study was to evaluate the changes of ECM proteins and of molecules responsible for matrix degradation induced by diabetic stimuli using a 3D valve leaflet model. The model was developed using a methacrylate gelatin hydrogel laden with human valvular interstitial cells (VICs) and seeded with a monolayer of human valvular endothelial cells (VECs) on the surface and was exposed to normal (NG) or high glucose (HG) conditions for 7 or 14 days. We evaluated: i) gene expression of ECM proteins by qRT-PCR for: collagen I, III, fibronectin, laminin, and remodeling enzymes MMP-1, -2, -9 and MMP13; ii) protein expression of ECM elements (collagen III, collagen IV and elastin) was quantified by western blot on cell lysate and immunofluorescence signal on 3D construct cryosections. Our data showed that exposure of the 3D construct to HG levels significantly
modified gene and protein expression of molecules involved in ECM remodeling, such us: collagens,
fibronectin, laminin and MMPs expression. In conclusion, our data suggest that diabetic conditions could induce an enhanced remodeling process in a 3D model of human aortic valve leaflet, feature that could explain the accelerated progression of CAVD in diabetic patients.
Acknowledgements: “this work was supported by a grant from the Competitiveness Operational Program 2014-2020, Targeted therapies for aortic valve disease in diabetes, THERAVALDIS, ID P_37_298, MySMIS code: 104362, contract number 115/13.09.2016”