Masjedi Shirin, Amarnath Adithi, Baily Katherine M, Ferdous Zannatul
Department of Mechanical, Aerospace, and Biomedical Engineering, The University of Tennessee, Knoxville, TN 37996.
Department of Mechanical, Aerospace, and Biomedical Engineering, The University of Tennessee, Knoxville, TN 37996.
Cardiovasc Pathol. 2016 May-Jun;25(3):185-194. doi: 10.1016/j.carpath.2015.12.002. Epub 2015 Dec 30.
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.
VICs were isolated from each of the aortic valve cusps and cultured in calcifying medium for 14days 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.
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.
钙化性主动脉瓣疾病(CAVD)是发达国家老年人中最常见的疾病之一。CAVD通过细胞介导的过程发展,临床数据表明CAVD主要始于主动脉瓣的无冠瓣。瓣膜间质细胞(VICs)存在于心脏瓣膜内部,是一种异质性细胞群体。本研究的目的是阐明从猪主动脉瓣的左、右和无冠瓣分离出的VICs在钙化潜能上的差异。
从每个主动脉瓣叶分离出VICs,并在钙化培养基中培养14天以诱导钙化。使用茜素红和冯科萨染色、碱性磷酸酶(ALP)染色、ALP酶活性测定和蛋白质免疫印迹法评估样本中的钙沉积、结节形成和钙化标志物。使用胶原蛋白和糖胺聚糖(GAG)测定法以及明胶酶谱法测量细胞外基质的产生和降解。我们观察到,与有冠瓣叶相比,从无冠瓣分离出的VICs表达上述钙化标志物的量最大。此外,无冠VICs中的胶原蛋白和GAG含量最高。然而,我们的酶谱结果显示,仅右VICs和无冠VICs之间的活性基质金属蛋白酶-2表达存在显著差异。
我们的结果表明,主动脉瓣三个瓣叶中的VICs可能本质上不同,其中一部分VICs可能易发生钙化。