Chondroitin Sulfate Promotes Interstitial Cell Activation and Calcification in an In Vitro Model of the Aortic Valve
Chondroitin Sulfate Promotes Interstitial Cell Activation and Calcification in an In Vitro Model of the Aortic Valve
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DOI：
10.1007/s13239-021-00586-z
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发表时间：
2021-11-04
影响因子：
1.8
通讯作者：
Mahler, Gretchen J.
中科院分区：
文献类型：
--
作者：
Dahal, Sudip;Bramsen, Jonathan Alejandro;Mahler, Gretchen J.
目的钙性主动脉瓣病(CAVD)是一种细胞级联改变，导致瓣叶增厚和瓣膜钙化。在病变的主动脉瓣中，通常在瓣膜海绵状组织中发现的糖胺多聚糖(GAG)迁移到钙化结节附近富含I型胶原的纤维层。目前对CAVD的治疗仅限于瓣膜置换术或针对其他心血管疾病量身定做的药物。方法将猪主动脉瓣间质细胞和猪主动脉瓣内皮细胞接种于不同初始硬度或初始硬度匹配的I型胶原水凝胶中，其中含有糖胺聚糖硫酸软骨素(CS)、透明质酸(HA)或硫酸皮肤素(DS)。培养2周后进行细胞基因表达、蛋白表达、蛋白分泌和钙化的检测。进行多元回归分析，以确定初始水凝胶硬度、GAG和内皮细胞的存在对钙化的重要性，无论是否使用成骨介质。结果高初始硬度水凝胶和成骨介质促进了钙化，而对于DS或HA，内皮细胞的存在阻止了钙化。研究发现，壳聚糖能增加促钙化基因的表达，增加激活的肌成纤维细胞蛋白的表达，诱导激活的间质细胞分泌I型胶原，并增加钙化结节的形成。结论本研究建立了一种较为完整的主动脉瓣病变模型，包括内皮细胞、间质细胞和僵硬的病态细胞外基质。健康瓣膜和病变瓣膜的体外模型有助于了解CAVD的发病机制，并为测试新的治疗方法提供了一个模型。
Purpose Calcific aortic valve disease (CAVD), has been characterized as a cascade of cellular changes leading to leaflet thickening and valvular calcification. In diseased aortic valves, glycosaminoglycans (GAGs) normally found in the valve spongiosa migrate to the collagen I-rich fibrosa layer near calcified nodules. Current treatments for CAVD are limited to valve replacement or drugs tailored to other cardiovascular diseases. Methods Porcine aortic valve interstitial cells and porcine aortic valve endothelial cells were seeded into collagen I hydrogels of varying initial stiffness or initial stiffness-matched collagen I hydrogels containing the glycosaminoglycans chondroitin sulfate (CS), hyaluronic acid (HA), or dermatan sulfate (DS). Assays were performed after 2 weeks in culture to determine cell gene expression, protein expression, protein secretion, and calcification. Multiple regression analyses were performed to determine the importance of initial hydrogel stiffness, GAGs, and the presence of endothelial cells on calcification, both with and without osteogenic medium. Results High initial stiffness hydrogels and osteogenic medium promoted calcification, while for DS or HA the presence of endothelial cells prevented calcification. CS was found to increase the expression of pro-calcific genes, increase activated myofibroblast protein expression, induce the secretion of collagen I by activated interstitial cells, and increase calcified nodule formation. Conclusion This study demonstrates a more complete model of aortic valve disease, including endothelial cells, interstitial cells, and a stiff and disease-like ECM. In vitro models of both healthy and diseased valves can be useful for understanding the mechanisms of CAVD pathogenesis and provide a model for testing novel therapeutics.