Bader A, Schilling T, Teebken O E, Brandes G, Herden T, Steinhoff G, Haverich A
Leibniz Research Laboratories for Biotechnology and Artificial Organs, LEBAO, Hannover Medical School, Germany.
Eur J Cardiothorac Surg. 1998 Sep;14(3):279-84. doi: 10.1016/s1010-7940(98)00171-7.
Tissue engineering of heart valves represents a new experimental concept to improve current modes of therapy in valvular heart disease. Drawbacks of glutaraldehyde fixed tissue valves or mechanical valves include the short durability or the need for life-long anticoagulation, respectively. Both have in common the inability to grow, which makes valvular heart disease especially problematic in children. The aim of this study was to develop a new methodology for a tissue engineered heart valve combining human cells and a xenogenic acellularized matrix.
Porcine aortic valves were acellularized by deterging cell extraction using Triton without tanning. Endothelial cells were isolated in parallel from human saphenous veins and expanded in vitro. Specimens of the surface of the acellular matrix were seeded with endothelial cells. Analysis of acellularity was performed by light microscopy and scanning electron microscopy. Cell viability following seeding was assayed by fluorescence staining of viable cells.
The acellularization procedure resulted in an almost complete removal of the original cells while the 3D matrix was loosened at interfibrillar zones. However the 3D arrangement of the matrix fibers was grossly maintained. The porcine matrix could be seeded with in vitro expanded human endothelial cells and was maintained in culture for up to 3 days to document the formation of confluent cultures.
Porcine aortic valves can be almost completely acellularized by a non-tanning detergent extraction procedure. The xenogenic matrix was reseeded with human endothelial cells. This approach may eventually lead to the engineering of tissue heart valves repopulated with the patients own autologous cells.
心脏瓣膜组织工程是一种新的实验概念，旨在改善目前治疗心脏瓣膜疾病的方式。戊二醛固定组织瓣膜或机械瓣膜的缺点分别是耐久性短或需要终身抗凝。两者的共同缺点是无法生长，这使得心脏瓣膜疾病在儿童中尤其成问题。本研究的目的是开发一种新的方法，用于构建一种结合人类细胞和异种脱细胞基质的组织工程心脏瓣膜。
使用曲拉通通过去垢剂细胞提取法对猪主动脉瓣进行脱细胞处理，不进行鞣制。同时从人隐静脉中分离内皮细胞并在体外扩增。将内皮细胞接种到脱细胞基质表面的标本上。通过光学显微镜和扫描电子显微镜进行脱细胞分析。接种后通过活细胞荧光染色测定细胞活力。
脱细胞处理几乎完全去除了原有的细胞，同时三维基质在纤维间区域变得疏松。然而，基质纤维的三维排列基本得以保留。猪基质可以接种体外扩增的人内皮细胞，并在培养中维持长达3天，以记录汇合培养物的形成。
猪主动脉瓣可以通过非鞣制去垢剂提取程序几乎完全脱细胞。异种基质重新接种了人内皮细胞。这种方法最终可能导致构建用患者自身自体细胞重新填充的组织心脏瓣膜。
