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In Vitro Generation of Atrioventricular Heart Valve Neoscaffolds

机译：房室心脏瓣膜新支架的体外生成

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Tissue engineering of cardiovascular structures represents a novel approach to improve clinical strategies in heart valve disease treatment. The aim of this study was to engineer decellularized atrioventricular heart valve neoscaffolds with an intact ultrastructure and to reseed them with umbilical cord-derived endothelial cells under physiological conditions in a bioreactor environment. Mitral (n=38) and tricuspid (n=36) valves were harvested from 40 hearts of German Landrace swine from a selected abattoir. Decellularization of atrioventricular heart valves was achieved by a detergent-based cell extraction protocol. Evaluation of the decellularization method was conducted with light microscopy and quantitative analysis of collagen and elastin content. The presence of residual DNA within the decellularized atrioventricular heart valves was determined with spectrophotometric quantification. The described decellularization regime produced full removal of native cells while maintaining the mechanical stability and the quantitative composition of the atrioventricular heart valve neoscaffolds. The surface of the xenogeneic matrix could be successfully reseeded with in vitro-expanded human umbilical cord-derived endothelial cells under physiological flow conditions. After complete decellularization with the detergent-based protocol described here, physiological reseeding of the xenogeneic neoscaffolds resulted in the formation of a confluent layer of human umbilical cord-derived endothelial cells. These results warrant further research toward the generation of atrioventricular heart valve neoscaffolds on the basis of decellularized xenogeneic tissue.
机译：心血管结构的组织工程学代表了一种改善心脏瓣膜疾病治疗临床策略的新颖方法。这项研究的目的是工程化具有完整的超微结构的脱细胞的房室心脏瓣膜新支架，并在生物反应器环境中，在生理条件下用脐带来源的内皮细胞重新植入它们。从选定屠宰场的40头德国长白猪中收获二尖瓣（n = 38）和三尖瓣（n = 36）。通过基于去污剂的细胞提取方案可实现房室心脏瓣膜的脱细胞。用光学显微镜对脱细胞方法进行评估，并对胶原蛋白和弹性蛋白含量进行定量分析。用分光光度法定量测定脱细胞的房室心脏瓣膜中残留DNA的存在。所描述的脱细胞机制可完全去除天然细胞，同时保持房室心脏瓣膜新支架的机械稳定性和定量组成。异种基质的表面可以在生理流动条件下与体外扩增的人脐带来源的内皮细胞成功地再播种。用此处描述的基于洗涤剂的方案完全脱细胞后，异种新支架的生理再接种导致人脐带血内皮细胞融合层的形成。这些结果值得进一步研究以脱细胞异种组织为基础的房室心脏瓣膜新支架的产生。

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