Platform technologies for decellularization, tunic-specific cell seeding, and in vitro conditioning of extended length, small diameter vascular grafts.
Platform technologies for decellularization, tunic-specific cell seeding, and in vitro conditioning of extended length, small diameter vascular grafts.
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DOI：
10.1089/ten.tec.2014.0047
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发表时间：
2014-12
期刊：
影响因子：
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通讯作者：
George Fercana;D. Bowser;Margarita Portilla;E. Langan;C. Carsten;D. Cull;L. Sierad;D. Simionescu
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文献类型：
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作者：
George Fercana;D. Bowser;Margarita Portilla;E. Langan;C. Carsten;D. Cull;L. Sierad;D. Simionescu
本研究的目的是产生长度较长、直径较小的血管支架，可作为治疗急性缺血的潜在移植物。生物组织被认为是优秀的支架，具有足够的生物、机械和操作性能；然而，它们在植入后往往会退化、扩张和钙化。我们假设化学稳定的无细胞动脉将是开发用于外周手术应用的血管移植物的理想支架。根据我们实验室和其他实验室的有希望的历史数据，我们选择对牛乳动脉和股动脉进行脱细胞，并测试它们作为血管移植支架的效果。这种长结构的脱细胞化需要开发一种新型的“生物加工”系统和一系列去污剂和酶，以产生完全无细胞的、半乳糖-(α1,3)-半乳糖(α-Gal)、无异种抗原的支架，并保留胶原蛋白、弹性蛋白和基底膜成分。脱细胞动脉表现出优异的机械性能，包括爆破压力、缝合线保持强度和弹性反冲力。为了减少弹性蛋白变性，我们用五没食子酰葡萄糖处理支架，然后使用外衣特异性细胞方法在体外使它们恢复活力。还开发了一种使用外部支架的新型无创伤内皮化方案，用于在流动生物反应器中调节的长移植物和细胞接种结构。脱细胞和再生都是可行的，但体外细胞保留仍然构成挑战。这些研究支持进一步努力临床使用小直径无细胞动脉作为血管移植物。
The aim of this study was to generate extended length, small diameter vascular scaffolds that could serve as potential grafts for treatment of acute ischemia. Biological tissues are considered excellent scaffolds, which exhibit adequate biological, mechanical, and handling properties; however, they tend to degenerate, dilate, and calcify after implantation. We hypothesized that chemically stabilized acellular arteries would be ideal scaffolds for development of vascular grafts for peripheral surgery applications. Based on promising historical data from our laboratory and others, we chose to decellularize bovine mammary and femoral arteries and test them as scaffolds for vascular grafting. Decellularization of such long structures required development of a novel "bioprocessing" system and a sequence of detergents and enzymes that generated completely acellular, galactose-(α1,3)-galactose (α-Gal) xenoantigen-free scaffolds with preserved collagen, elastin, and basement membrane components. Acellular arteries exhibited excellent mechanical properties, including burst pressure, suture holding strength, and elastic recoil. To reduce elastin degeneration, we treated the scaffolds with penta-galloyl glucose and then revitalized them in vitro using a tunic-specific cell approach. A novel atraumatic endothelialization protocol using an external stent was also developed for the long grafts and cell-seeded constructs were conditioned in a flow bioreactor. Both decellularization and revitalization are feasible but cell retention in vitro continues to pose challenges. These studies support further efforts toward clinical use of small diameter acellular arteries as vascular grafts.