ARTIFICIAL EXTRACELLULAR MATRIX PROTEINS FOR VASCULAR GR

这项工作的长期目标是开发新的基因工程材料，用于制造以改善长期通畅性为特征的血管移植物。该方法基于以下假设：i）。 现有的合成血管移植物材料-特别是膨胀型聚四氟乙烯（ePTFE）和涤纶-在表面化学或机械性能方面都不是最佳的，ii）。合成移植物的愈合机制至少部分地由与移植物表面或与沉积在该表面上的大分子（包括血浆蛋白或细胞外基质（ECM）蛋白）的细胞相互作用控制，iii）.移植物表面细胞行为的控制措施可以通过呈递特异性细胞表面受体的配体来获得，和iv）。 ECM蛋白的工程化变体将允许控制移植物的关键机械性质和移植物表面配体的呈递。该项目将包括：i）。微生物表达编码掺入特异性细胞结合和交联结构域的人工ECM蛋白的人工基因，ii）.由这些蛋白质制备的交联膜的制造和机械表征，iii）.测定这些表面相对于在生理学相关剪切应力下培养的内皮细胞的粘附性，和iv）. 分析在人工细胞外基质上培养的内皮细胞单层的行为。

The long-term objective of this work is the development of new genetically engineered materials to be used in the fabrication of vascular grafts characterized by improving long-term patency. The approach is predicated on the following hypotheses: i). that existing synthetic vascular graft materials - specifically expanded polytetrafluoroethylene (ePTFE) and Dacron - are not optimal, either in terms of surface chemistry or with respect to mechanical properties, ii). that the mechanisms of healing of synthetic grafts are controlled at least in part by cellular interactions with the graft surface or with macromolecules, including plasma proteins or extracellular matrix (ECM) proteins, deposited on that surface, iii). that a measure of control of cellular behavior at the graft surface can be gained by presentation of ligands for specific cell- surface receptors, and iv). that engineered variants of ECM proteins will allow control, both of the key mechanical properties of the graft, and of the presentation of ligands at the graft surface. The project will include: i). microbial expression of artificial genes encoding artificial ECM proteins that incorporate specific cell-binding and crosslinking domains, ii). fabrication and mechanical characterization of crosslinked films prepared from these proteins, iii). determination of the adhesivity of these surfaces with respect to endothelial cells cultured under physiologically relevant shear stresses, and iv). analysis of the behavior of endothelial cell monolayers cultured on artificial extracellular matrices.