Comparison of calcification potential of valvular interstitial cells isolated from individual aortic valve cusps

Background: Calcific aortic valve disease (CAVD) is one of the most prevalent disorders among the elderly in developed countries. CAVD develops via cell-mediated processes, and clinical data show that CAVD initiates mostly in the noncoronary cusp of the aortic valve. Valvular interstitial cells (VICs) populate the inside of heart valves and are a heterogeneous cell population. The goal of this study is to elucidate the difference in calcification potential among VICs isolated from the left, right, and noncoronary cusps of porcine aortic valves. Methods and results: VICs were isolated from each of the aortic valve cusps and cultured in calcifying medium for 14 days to induce calcification. The samples were assessed for calcium deposits, nodule formation, and calcific markers using alizarin red and Von Kossa staining, alkaline phosphatase (ALP) staining, ALP enzyme activity assay, and Western blot. Extracellular matrix production and degradation were measured using collagen and glycosaminoglycan (GAG) assay and gelatin zymography. We observed that VICs isolated from the noncoronary cusp expressed greatest amount of the above calcific markers as compared to the coronary cusps. Also, collagen and GAG content was the greatest in noncoronary VICs. However, our zymography results showed significant difference only for active matrix metalloproteinase-2 expression between right and noncoronary VICs. Conclusion: Our results suggest that VICs among the three cusps within aortic valve might be inherently different, where a subpopulation of VICs might be predisposed to calcification. (C) 2015 Elsevier Inc. All rights reserved.