Thrombosis Research Unit, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Poojapura, Trivandrum 695 012, India.
Biofabrication. 2010 Dec;2(4):041001. doi: 10.1088/1758-5082/2/4/041001. Epub 2010 Sep 24.
Porous and pliable conduits made of biodegradable polymeric scaffolds offer great potential for the development of blood vessel substitutes but they generally lack signals for cell proliferation, survival and maintenance of a normal phenotype. In this study we have prepared and evaluated porous poly(ε-caprolactone) (PCL) integrated with fibrin composite (FC) to get a biomimetic hybrid scaffold (FC PCL) with the biological properties of fibrin, fibronectin (FN), gelatin, growth factors and glycosaminoglycans. Reduced platelet adhesion on a human umbilical vein endothelial cell-seeded hybrid scaffold as compared to bare PCL or FC PCL was observed, which suggests the non-thrombogenic nature of the tissue-engineered scaffold. Analysis of real-time polymerase chain reaction (RT-PCR) after 5 days of endothelial cell (EC) culture on a hybrid scaffold indicated that the prothrombotic von Willebrand factor and plasminogen activator inhibitor (PAI) were quiescent and stable. Meanwhile, dynamic expressions of tissue plasminogen activator (tPA) and endothelial nitric oxide synthase indicated the desired cell phenotype on the scaffold. On the hybrid scaffold, shear stress could induce enhanced nitric oxide release, which implicates vaso-responsiveness of EC grown on the tissue-engineered construct. Significant upregulation of mRNA for extracellular matrix (ECM) proteins, collagen IV and elastin, in EC was detected by RT-PCR after growing them on the hybrid scaffold and FC-coated tissue culture polystyrene (FC TCPS) but not on FN-coated TCPS. The results indicate that the FC PCL hybrid scaffold can accomplish a remodeled ECM and non-thrombogenic EC phenotype, and can be further investigated as a scaffold for cardiovascular tissue engineering.
多孔且柔韧的生物可降解聚合物支架制成的导管在开发血管替代物方面具有巨大的潜力，但它们通常缺乏促进细胞增殖、存活和维持正常表型的信号。在这项研究中，我们制备和评估了多孔聚（ε-己内酯）（PCL）与纤维蛋白复合（FC）的集成，以获得具有纤维蛋白、纤维连接蛋白（FN）、明胶、生长因子和糖胺聚糖的仿生杂化支架（FC-PCL）。与裸 PCL 或 FC-PCL 相比，在人脐静脉内皮细胞接种的杂化支架上观察到血小板黏附减少，这表明组织工程支架具有非血栓形成特性。对内皮细胞（EC）在杂化支架上培养 5 天后进行实时聚合酶链反应（RT-PCR）分析表明，促血栓形成的血管性血友病因子和纤溶酶原激活物抑制剂（PAI）处于静止和稳定状态。同时，组织型纤溶酶原激活物（tPA）和内皮型一氧化氮合酶的动态表达表明支架上具有所需的细胞表型。在杂化支架上，剪切力可诱导一氧化氮释放增强，这表明在组织工程构建物上生长的 EC 具有血管反应性。通过 RT-PCR 检测到，在杂化支架和 FC 涂层的组织培养聚苯乙烯（FC-TCPS）上培养 EC 后，细胞外基质（ECM）蛋白、IV 型胶原和弹性蛋白的 mRNA 表达显著上调，但在 FN 涂层的 TCPS 上则没有。结果表明，FC-PCL 杂化支架可以完成重塑的 ECM 和非血栓形成的 EC 表型，并且可以进一步作为心血管组织工程的支架进行研究。
