Chronic High Glucose Concentration Induces Inflammatory and Remodeling Changes in Valvular Endothelial Cells and Valvular Interstitial Cells in a Gelatin Methacrylate 3D Model of the Human Aortic Valve.

Monica Madalina Tucureanu,Ionel Droc,Razvan Daniel Macarie,Sergiu Cecoltan,Ileana Manduteanu,Mihaela Vadana,Andreea Cristina Mihaila,Letitia Ciortan,Elena Butoi

doi:10.3390/polym12122786

Monica Madalina Tucureanu, Ionel Droc + Show 7 more

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https://doi.org/10.3390/polym12122786

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Journal: Polymers	Publication Date: Nov 25, 2020
Citations: 19	License type: CC BY 4.0

Affiliation: Clinical Emergency Hospital Bucharest

Abstract

Calcific aortic valve disease (CAVD), a degenerative disease characterized by inflammation, fibrosis and calcification, is accelerated in diabetes. Hyperglycemia contributes to this process by mechanisms that still need to be uncovered. We have recently developed a 3D model of the human aortic valve based on gelatin methacrylate and revealed that high glucose (HG) induced osteogenic molecules and increased calcium deposits in a pro-osteogenic environment. To further understand the events leading to calcification in diabetic conditions in CAVD, we analyzed here the inflammatory and remodeling mechanisms induced by HG in our 3D model. We exposed valvular endothelial cells (VEC) and interstitial cells (VIC) to normal glucose (NG) or HG for 7 and 14 days, then we isolated and separated the cells by anti-CD31 immunomagnetic beads. The changes induced by HG in the 3D model were investigated by real-time polymerase chain reaction (RT-PCR), Western blot, enzyme-linked immunosorbent assay (ELISA) and immunofluorescence. Our results showed that HG induced expression of different cytokines, cell adhesion molecules and matrix metalloproteinases in VEC and VIC. In addition, protein kinase C was increased in VEC and VIC, indicating molecular mechanisms associated with HG induced inflammation and remodeling in both valvular cells. These findings may indicate new biomarkers and targets for therapy in diabetes associated with CAVD.

Highlights

Calcific aortic valve disease (CAVD) is a degenerative disease characterized by inflammation, fibrosis and calcification, leading to the hardening of the aortic valve leaflet, orifice narrowing and stenosis
We found that after 7 days MCP-1 was significantly increased by High Glucose (HG) both in valvular endothelial cells (VEC) and in valvular interstitial cells (VIC), while at day14, MCP-1 expression was found at similar levels as in controls in VEC and at a slightly but not significantly increased level in VIC
Is it known that CAVD is accelerated in diabetes, the early mechanisms leading to aortic valve calcification still need to be uncovered

Summary

Introduction

Calcific aortic valve disease (CAVD) is a degenerative disease characterized by inflammation, fibrosis and calcification, leading to the hardening of the aortic valve leaflet, orifice narrowing and stenosis. Studies that investigated the influence of DM on patients with CAVD [2] showed that it can influence the natural course and increases the progression rate of AS [3,4,5]. Accumulating data indicate that AS is an active process similar to atherosclerosis involving lipid infiltration, inflammation, osteoblastic transition of valve interstitial cells and active leaflet calcification [6]. The initial stage of degenerative AS has more similarities with atherosclerosis than the progressive phase, where fibrosis and calcification are more prominent. The connection between lipids and the development of degenerative AS was presumed and sustained by detection of diffuse atherosclerotic lesions in the aortic leaflets of patients with familial hypercholesterolemia and no other atherosclerotic risk factors [8]. The contributing role of elevated LDL (low-density lipoproteins) and the risk of both vascular atherosclerosis and AS was presented in the Cardiovascular Health Study [9]

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