High Glucose Induced Changes in Human VEC Phenotype in a 3D Hydrogel Derived From Cell-Free Native Aortic Root

Abstract

Background: Valvular endothelial cells (VEC) have key roles in maintaining valvular integrity and homeostasis, and dysfunctional VEC are the initiators and major contributors to aortic valve disease in diabetes. Previous studies have shown that HG stimulated an inflammatory phenotype in VEC. Inflammation was shown to induce endothelial to mesenchymal transition (EndMT), a process extensively involved in many pathologies, including calcification of the aortic valve. However, the effect of HG on EndMT in VEC is not known. In addition, there is evidence that endothelin (ET) is a proinflammatory agent in early diabetes and was detected in aortic stenosis, but it is not known whether HG induces ET and endothelin receptors and whether endothelin modulates HG-dependent inflammation in VEC. This study aims to evaluate HG effects on EndMT, on endothelin and endothelin receptors induction in VEC and their role in HG induced VEC inflammation.Methods and Results: We developed a new 3D model of the aortic valve consisting of a hydrogel derived from a decellularized extracellular cell matrix obtained from porcine aortic root and human valvular cells. VEC were cultured on the hydrogel surface and VIC within the hydrogel, and the resulted 3D construct was exposed to high glucose (HG) conditions. VEC from the 3D construct exposed to HG exhibited: attenuated intercellular junctions and an abundance of intermediate filaments (ultrastructural analysis), decreased expression of endothelial markers CD31 and VE–cadherin and increased expression of the mesenchymal markers α-SMA and vimentin (qPCR and immunocytochemistry), increased expression of inflammatory molecules ET-1 and its receptors ET-A and ET-B, ICAM-1, VCAM-1 (qPCR and Immunocytochemistry) and augmented adhesiveness. Blockade of ET-1 receptors, ET-A and ET-B reduced secretion of inflammatory biomarkers IL-1β and MCP-1 (ELISA assay).Conclusions: This study demonstrates that HG induces EndMT in VEC and indicates endothelin as a possible target to reduce HG-induced inflammation in VEC.

Major Facilitator Superfamily Domain Containing 5 Inhibition Reduces Lipoprotein(a) Uptake and Calcification in Valvular Heart Disease

“…expression of procalcific proteins, VECs can undergo an endothelial-to-mesenchymal transformation into osteogenic cells, 39 and high glucose stimulates endothelial-tomesenchymal transformation of VECs. 40 Our current TriCEPS ligand-receptor capture affinity purification results support MFSD5 binding to Lp(a) and implicate a functional role in a lipid metabolism. It remains unclear what portion of Lp(a) that MFSD5 interacts with, but it is unlikely to be ApoB given that MFSD5 siRNA did not reduce LDL uptake in vitro.…”

New insights into the pathogenesis of cardiac papillary fibroelastomas

“…showed an increased abundance of intermediate filaments and attenuation of intercellular junctions. Besides ultrastructural results, higher expression of mesenchymal markers, such as vimentin and α-SMA, indicated on the participation of high glucose conditions in EndMT process via VEC inflammation [23]. In addition, secretion of extracellular vesicles by endocardial endothelial cells into the extracellular space may be, in part, attributable to the remote regulatory function of these cells.…”

“…The process of the endocardial endothelium transition has an important role in the endocardial cushion formation during heart valve development [20,[42][43][44]. In addition, recent reports indicated that endothelial-to-mesenchymal transition may also occur in postnatal valves in response to injury and contributes to the development of various cardiovascular diseases, including myocardial infarction, cardiac fibrosis, valve calcification, endocardial elastofibrosis, atherosclerosis, and pulmonary arterial hypertension [12,23,41,45].…”

Investigation of biological and wound healing effects of Estrogen solution: An in vitro study

“…mRNA levels of CD31 and Vimentin were determined using the SYBR1 Premix Ex TaqTM II Kit (Tli RNaseH Plus, Chin). β-actin was used as an internal housekeeping gene in co-cultured cells, and the mRNA relative expression of the CD31 and Vimentin genes was quanti ed by the 2 −∆∆Ct method [20]. The primers used for Real-Time PCR analysis are listed in Table 1.…”