Inserm U577, Bordeaux and Université Victor Segalen Bordeaux 2 F-33076, France.
Acta Biomater. 2010 Apr;6(4):1437-45. doi: 10.1016/j.actbio.2009.11.012. Epub 2009 Nov 12.
There is considerable interest in making multilayer films for various applications, among which are cell contacting biomaterials, allowing new opportunities to prepare functionalized biomaterials. In this study we have explored the capability of poly(sodium-4-styrene sulfonate)/poly(allylamine hydrochloride) polyelectrolyte multilayer films (PMFs) as functional coatings for human progenitor-derived endothelial cells (PDECs), since the latter are a potential source of endothelial-type cells to be used in bioartificial vascular substitutes. We performed investigations with PDECs derived from peripheral blood and characterized as endothelial cells. After forming a confluent monolayer on PMFs they were exposed to laminar pulsatile physiological shear stress. We investigated whether PDECs were able to withstand shear stress and to respond at the mRNA (microarray analysis) and protein levels (thrombomodulin and tissue factor functional activity), in comparison with collagen I and fibrin glue used as controls. After shear stress the PDECs remained spread on the substrates, with a resulting increase in the number of expressed genes. Considering the functional significance of our findings for the regulation of coagulation and fibrinolytic factors, mRNA tissue plasminogen activator and thrombomodulin, profibrinolytic and thrombin inhibiting respectively, were overexpressed in PDECs after 6h shear stress. von Willebrand factor showed down-regulation, while tissue factor was up-regulated. We can speculate that PMFs could favour anti-thrombogenic activity by PDECs because activated protein C generation, measuring thrombomodulin activity, was particularly high on PMFs, but unchanged after 6h shear stress. Thus, PMFs could represent suitable coatings able to provide functional surfaces for endothelialization with PDECs.
人们对用于各种应用的多层膜非常感兴趣，其中包括与细胞接触的生物材料，这为制备功能化生物材料提供了新的机会。在这项研究中，我们探索了聚（4-苯乙烯磺酸钠）/聚（盐酸烯丙胺）聚电解质多层膜（PMF）作为人原代内皮祖细胞（PDEC）功能涂层的能力，因为后者是内皮型细胞的潜在来源，可用于生物人工血管替代品。我们用来源于外周血的并已被鉴定为内皮细胞的 PDEC 进行了研究。在 PMF 上形成致密单层后，将其暴露于层流脉动生理剪切力下。我们研究了 PDEC 是否能够承受剪切力，并在 mRNA（微阵列分析）和蛋白水平（血栓调节蛋白和组织因子功能活性）上做出反应，与用作对照的胶原 I 和纤维蛋白胶进行比较。在经受剪切力后，PDEC 仍在基质上展开，导致表达基因的数量增加。考虑到我们的发现对凝血和纤维蛋白溶解因子调节的功能意义，mRNA 组织纤溶酶原激活物和血栓调节蛋白，分别具有抗纤维蛋白溶解和抗凝血酶作用，在经受 6 小时剪切力后在 PDEC 中过度表达。血管性血友病因子显示下调，而组织因子上调。我们可以推测，PMF 可以通过 PDEC 促进抗血栓形成活性，因为测量血栓调节蛋白活性的活化蛋白 C 的产生在 PMF 上特别高，但在 6 小时剪切力后没有变化。因此，PMF 可以代表合适的涂层，能够为 PDEC 内皮化提供功能表面。
