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

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.

Crosstalk of Diabetic Conditions with Static Versus Dynamic Flow Environment—Impact on Aortic Valve Remodeling

“…Others have reported the induction of SPP1 and TGFβ in 3D models cultivating VIC under HG treatment. However, VIC in the latter model have been co-cultured with valvular endothelial cells [31]. Discrepancies between this model and our models thus might not only be due to mechanical differences but also might also be ascribed to the lack of VEC in our models, since VEC are known to communicate with VIC [32] possibly through protein kinase C under HG [24].…”

Nano-Polyplexes Mediated Transfection of Runx2-shRNA Mitigates the Osteodifferentiation of Human Valvular Interstitial Cells

“…Human VIC were isolated from noncalcified cusps (or portions of the cusp) of aortic valves obtained from a patient who underwent surgical valve replacement, as recently described [ 26 ]. The surgery was carried out at Central Military Hospital “Dr.…”

“…Human VIC were isolated from noncalcified cusps (or portions of the cusp) of aortic valves obtained from a patient who underwent surgical valve replacement, as recently described [26]. The surgery was carried out at Central Military Hospital "Dr. Carol Davila", Cardiovascular surgery clinic, Bucharest, according to the principles outlined in the Declaration of Helsinki for experiments involving human samples [27].…”

Reproducible In Vitro Tissue Culture Model to Study Basic Mechanisms of Calcific Aortic Valve Disease: Comparative Analysis to Valvular Interstitials Cells

“…The co-culturing techniques also have some disadvantages, because maintenance of the quality and stability of the population of co-cultured species is a tedious task [ 28 ]. However, recently described novel 3D models with human VICs, which are suitable to investigate VIC phenotype changes as a result of both communication with valvular endothelial cells and exposure to pathological stimuli, provide promising tools to better understand the valve cell biology and pathological mechanisms underlying CAVD [ 29 , 30 ].…”