Endothelial to Mesenchymal Transformation-derived Activated Fibroblast Behavior in a 3D Culture Environment
Endothelial to Mesenchymal Transformation-derived Activated Fibroblast Behavior in a 3D Culture Environment
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3D 培养环境中内皮细胞向间充质转化衍生的激活成纤维细胞行为
DOI：
10.1080/24748706.2021.1901523
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Mahler, Gretchen
中科院分区：
文献类型：
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作者：
Bramsen, Jonathan Alejandro;Alber, Bridget;Murray, Bruce;Chen, Mei-Hsiu;Huang, Peter;Mahler, Gretchen
目的：在钙化性主动脉瓣疾病（CAVD）中，糖胺聚糖（GAG）如硫酸软骨素（CS）和透明质酸（HA）存在于钙化结节附近的正常富含胶原的主动脉瓣纤维瘤中。先前使用3D体外模型的工作表明，GAG也可以促进内皮细胞向间充质细胞转化（EndMT）。间充质转化在CAVD进展中的作用尚不清楚。本研究的目的是调查的作用，糖胺聚糖和EndMT对瓣膜细胞的行为在一个三维，在体外模型的健康和患病valves.Methods：三维体外培养，代表健康（胶原1水凝胶）或患病（胶原1水凝胶+糖胺聚糖）瓣膜组织，接种猪主动脉瓣内皮细胞（PAVEC）和猪主动脉瓣间质细胞（PAVIC）。细胞培养物在常规培养基中生长14天，然后评价钙化结节形成、碱性磷酸酶活性、增殖和与EndMT活性相关的蛋白表达。含有CS条件的水凝胶诱导了最大的钙化，如用Alzarin Red定量所测量的，而含有HA条件的凝胶没有与对照组显著不同的钙化。增殖结果表明，CS条件下PAVEC和PAVIC均具有高水平的细胞分裂活性。蛋白质表达分析表明，增加EndMT水凝胶含有GAGs.Conclusions：三维，在体外模型的主动脉瓣模拟进展到晚期CAVD。进一步研究EndMT在主动脉瓣微环境中的下游作用对于随后改善治疗方案至关重要。
Objective: In calcific aortic valve disease (CAVD), glycosaminoglycans (GAGs) such as chondroitin sulfate (CS) and hyaluronic acid (HA) are present in the normally collagen-rich aortic valve fibrosa near calcified nodules. Previous work using a 3D in vitro model has shown that GAGs can also promote endothelial to mesenchymal transformation (EndMT). The role of mesenchymal transformation in the progression of CAVD remains unclear. The goal of this study is to investigate the role of GAGs and EndMT on valve cell behavior in a 3D, in vitro model of healthy and diseased valves.Methods: Three dimensional in-vitro cultures, representing healthy (collagen 1 hydrogels) or diseased (collagen 1 hydrogels + GAGs) valve tissue, were seeded with Porcine Aortic Valve Endothelial Cells (PAVEC) and Porcine Aortic Valve Interstitial Cells (PAVIC). Cell cultures were grown for 14 days in regular medium and then evaluated for calcified nodule formation, alkaline phosphatase activity, proliferation and protein expression relating to EndMT activityResults: Introduction of GAGs into the in vitro model resulted in decreased alkaline phosphatase activity. Hydrogels containing CS conditions induced the greatest calcification, as measured with Alzarin Red quantification, while gels containing HA conditions did not have calcification that was significantly different than controls. Proliferation results suggest high levels of cell division activity, for both PAVEC and PAVIC in CS conditions. Protein expression analysis indicated increased EndMT in hydrogels containing GAGs.Conclusions: 3D, in vitro models of the aortic valve simulated progression toward late stage CAVD. Further investigation of the downstream role of EndMT within the aortic valve microenvironment is critical to subsequent improvement in treatment options.