Molecular mechanisms involved in high glucose-induced valve calcification in a 3D valve model with human valvular cells.

Mihaela Vadana,Letitia Ciortan,Ileana Manduteanu,Andreea C Mihaila,Razvan D Macarie,Ionel Droc,Sergiu Cecoltan,Monica M Tucureanu,Elena Butoi

doi:10.1111/jcmm.15277

Mihaela Vadana, Letitia Ciortan + Show 7 more

Open Access

https://doi.org/10.1111/jcmm.15277

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Abstract

Calcific aortic valve disease (CAVD)—the most common valvular heart disease—is accelerated in diabetes and has no pharmacotherapy. Although it is known that early CAVD is associated with inflammation and osteogenesis, the molecular mechanisms involved in diabetes‐associated CAVD still need to be uncovered. In this context, we have developed a 3D construct based on gelatin populated with human valvular endothelial cells (VEC) and valvular interstitial cells (VIC) and evaluated the effect of high glucose (HG) concentration on osteogenic molecules expression and on calcification mechanisms. First, we characterized the 3D model and assessed VIC remodelling properties at different time‐points. Then, we exposed it to normal glucose (NG) or high glucose (HG) for 7, 14 and 21 days after which the cells were isolated, separated and investigated individually. Our results showed that encapsulated VIC actively remodel the hydrogel, as demonstrated by an increased expression of extracellular matrix (ECM) proteins and matrix metalloproteinases (MMPs). Moreover, exposure of the construct to HG triggered bone morphogenetic protein (BMP) and TGF‐β signalling pathways, up‐regulating expression of osteogenic molecules—BMP‐2/‐4, osteocalcin, osteopontin, SMADs and Runt‐related transcription factor (Runx‐2)—and increased calcium deposits in an osteogenic environment. These findings underline the potential of the developed 3D model as a suitable system to investigate the mechanisms of human CAVD and may help to better understand the calcification mechanisms in CAVD associated to diabetes.

Highlights

Aortic valve (AV) stenosis due to calcific aortic valve disease (CAVD) is the most common heart valve disease in developed countries.[1]
Calcific aortic valve disease is a degenerative disease characterized by inflammation, fibrosis and calcification, leading to hardening
In this study we developed a 3D model for aortic valve leaflet and exposed it to high glucose (HG) concentration, in order to investigate the mechanism of CAVD in a diabetic environment

Summary

| INTRODUCTION

Aortic valve (AV) stenosis due to calcific aortic valve disease (CAVD) is the most common heart valve disease in developed countries.[1]. Our group showed that HG induces enhanced monocyte adhesion to VEC via a mechanism involving the cell adhesion molecules: ICAM-1, VCAM-1 and CD 18.20 Recently, using a 2D culture, it was found that VIC from sheep origin do not show morphological changes and do not acquire an osteogenic phenotype in hyperinsulinaemia or hyperglycaemia.[21]. In this context, a 3D scaffold with human VEC and VIC, that supports and promotes VIC-mediated ECM remodelling, while preserving VIC fibroblastic phenotype, would be promising for the study of valvular cells interaction in normal as well as pathological conditions, and molecular mechanisms of leaflet calcification. These molecules were expressed differently in VIC versus VEC, suggesting both similar and different mechanisms of HG effects on valvular cells

| MATERIALS AND METHODS

Findings

| DISCUSSION