Co-expression of elastin and collagen leads to highly compliant engineered blood vessels
Co-expression of elastin and collagen leads to highly compliant engineered blood vessels
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DOI：
10.1002/jbm.a.32028
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发表时间：
2008-06-15
影响因子：
4.9
通讯作者：
Wang, Yadong
中科院分区：
文献类型：
--
作者：
Gao, Jin;Crapo, Peter;Wang, Yadong
弹性蛋白的合成和生理顺应性是血管组织工程的重要挑战。在这里，我们报告说，一个生物相容性的弹性支架可以支持弹性蛋白和胶原蛋白的共表达，这可能产生的结构中的生理顺应性。一种可生物降解的弹性体，聚（甘油癸二酸酯），制成高度多孔的管状支架。将原代狒狒动脉平滑肌细胞（SMC）接种在支架的管腔中，然后在温和灌注下培养1周。从狒狒外周血中分离的循环内皮祖细胞（EPCs）直接接种在第8天的管腔平滑肌层。在表征之前，使用脉动流系统再灌注构建体2周。在另一组实验中，将SMC培养7周并与EPC共培养1周。使用任一组培养条件获得的构建体含有弹性蛋白和胶原蛋白：Masson三色染色显示构建体中的环形胶原蛋白带，并且弹性蛋白从其特征性自发荧光、Verhoff染色和热碱消化后不溶性残余物的氨基酸分析中是明显的。所有构建体具有汇合的细胞腔，其中细胞良好分散在整个支架中。在生理压力下，8周结构的顺应性与压力-直径试验中观察到的人体动脉相当。弹性支架、EPC和SMC的共培养以及机械调节的组合似乎鼓励更天然的细胞外基质的表达并导致生理相关的顺应性;这两者都是血管组织工程中的主要挑战。(C)2008 Wiley Periodicals，Inc.
Elastin synthesis and physiologic compliance are significant challenges in blood vessel tissue engineering. Here, we report that a biocompatible elastomeric scaffold can support the co-expression of elastin and collagen, which likely yielded the physiologic compliance in the constructs. A biodegradable elastomer, poly(glycerol sebacate), was fabricated into highly porous tubular scaffolds. Primary baboon arterial smooth muscle cells (SMCs) were seeded in the lumen of the scaffolds followed by a 1-week culture under gentle perfusion. Circulating endothelial progenitor cells (EPCs) isolated from baboon peripheral blood was seeded directly on the smooth muscle layer in the lumen on day 8. The constructs were perfused using a pulsatile flow system for another 2 weeks before characterization. In another set of experiments, the SMCs were cultured for 7 weeks and were co-cultured for 1 week with the EPCs. Constructs obtained using either set of culture conditions contained elastin and collagen: Masson's trichrome stain showed a circumferential collagen band in the constructs, and elastin was evident from its characteristic autofluorescence, Verhoff's stain, and amino acid analysis of insoluble remnants after hot alkali digestion. All constructs had a confluent cellular lumen with cells well-dispersed throughout the scaffolds. At physiologic pressures, the compliance of the 8-week construct was comparable to human arteries as observed in pressure-diameter testing. Combination of elastomeric scaffolds, co-culture of EPC and SMC, and mechanical conditioning appears to encourage the expression of a more natural extracellular matrix and lead to physiologically-relevant compliance; both are major challenges in blood vessel tissue engineering. (C) 2008 Wiley Periodicals, Inc.