Department for Tissue Engineering & Textile Implants, Institute of Applied Medical Engineering, Helmholtz Institute of the RWTH Aachen University Hospital, Pauwelsstraße 20, 52074 Aachen, Germany.
Biomed Eng Online. 2013 Jan 28;12:7. doi: 10.1186/1475-925X-12-7.
Extracorporeal membrane oxygenation (ECMO) can replace the lungs' gas exchange capacity in refractory lung failure. However, its limited hemocompatibility, the activation of the coagulation and complement system as well as plasma leakage and protein deposition hamper mid- to long-term use and have constrained the development of an implantable lung assist device. In a tissue engineering approach, lining the blood contact surfaces of the ECMO device with endothelial cells might overcome these limitations. As a first step towards this aim, we hypothesized that coating the oxygenator's gas exchange membrane with proteins might positively influence the attachment and proliferation of arterial endothelial cells.
Sheets of polypropylene (PP), polyoxymethylpentene (TPX) and polydimethylsiloxane (PDMS), typical material used for oxygenator gas exchange membranes, were coated with collagen, fibrinogen, gelatin or fibronectin. Tissue culture treated well plates served as controls. Endothelial cell attachment and proliferation were analyzed for a period of 4 days by microscopic examination and computer assisted cell counting.
Endothelial cell seeding efficiency is within range of tissue culture treated controls for fibronectin treated surfaces only. Uncoated membranes as well as all other coatings lead to lower cell attachment. A confluent endothelial cell layer develops on fibronectin coated PDMS and the control surface only.
Fibronectin increases endothelial cells' seeding efficiency on different oxygenator membrane material. PDMS coated with fibronectin shows sustained cell attachment for a period of four days in static culture conditions.
体外膜肺氧合(ECMO)可以代替肺衰竭时肺部的气体交换能力。然而,其有限的血液相容性、凝血和补体系统的激活以及血浆渗漏和蛋白质沉积阻碍了其在中期至长期的使用,并限制了植入式肺辅助装置的发展。在组织工程方法中,用内皮细胞覆盖 ECMO 装置的血液接触面可能会克服这些限制。作为实现这一目标的第一步,我们假设用蛋白质涂覆氧合器的气体交换膜可能会对动脉内皮细胞的附着和增殖产生积极影响。
用胶原蛋白、纤维蛋白原、明胶或纤维连接蛋白涂覆典型的氧合器气体交换膜用的聚丙烯(PP)、聚氧化甲烯(TPX)和聚二甲基硅氧烷(PDMS)片。组织培养处理过的孔板作为对照。通过显微镜检查和计算机辅助细胞计数分析内皮细胞附着和增殖 4 天。
只有在用纤维连接蛋白处理的表面上,内皮细胞接种效率在组织培养处理对照范围内。未涂层的膜以及所有其他涂层都会导致附着细胞减少。只有在纤维连接蛋白涂覆的 PDMS 和对照表面上才能形成连续的内皮细胞层。
纤维连接蛋白可提高不同氧合器膜材料上内皮细胞的接种效率。在静态培养条件下,纤维连接蛋白涂覆的 PDMS 可维持 4 天的持续细胞附着。