Toward Development of in-situ Endothelialized Artificial Graft : Hydrodynamic Shear Stress Dependent Retention of Endothelial Cells Adhered to a Vascular Endothelial Growth Factor-Fixed Surface

原位内皮化人工移植物的开发：粘附于血管内皮生长因子固定表面的内皮细胞的流体动力剪切应力依赖性保留

基本信息

批准号：
18300165
负责人：
MATSUDA Takehisa
金额：
$ 10.93万
依托单位：
Kanazawa Institute of Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18300165/
关键词：
Biomaterial Cellular tissue Artficial blood vessel biomechanics

项目摘要

The full endothelialization of the luminal surface of an implanted small-diameter artificial graft is a requirement for expressing nonthrombogenic potential. Our strategic approach to meet that requirement is in-situ harvesting of circulating endothelial progenitor cells (EPCs) on a luminal surface of an implanted graft. The first phase of this research emphasized the determination of an appropriate receptor-counter-receptor pair, thereby enabling the clonal adhesion of EPC and high cell retention under flow loading. The radial flow chamber, installed with a three-regiospecific-protein-fixed substrate (fibronectin for integrin, and VEGF and anti-Flk-1 antibodies for VEGF receptor), clearly differentiated shear-stress-dependent cell retention potential of endothelial cells (ECs; ECs were used here as a cell model closed to EPCs because of difficulty in harvesting EPCs), which depends on the type of fixed protein and cell adhesion period. High shear-stress-resistance potential was in the order of fibronectin > VEGF and anti-Flk-1 antibody. The adhesion potential from the cell adhesion area and cell shape were both determined from phase-contrast microscopic and scanning electron microscopic images. The distribution and magnitude of fluorescence intensity of fluorescently immunostained vinculin (a key protein of focal adhesion plaque) were determined using total internal reflection fluorescence microscopy. All indicated that round cells were much more easily detachable than well-spread shaped cells. A potential in-situ EPC harvest technology is discussed in this paper.

植入的小直径人工移植物的管腔表面的完全内皮化是表达非血栓形成潜力的必要条件。我们满足这一要求的战略方法是在植入移植物的管腔表面原位收获循环内皮祖细胞 (EPC)。这项研究的第一阶段强调确定合适的受体-反受体对，从而实现 EPC 的克隆粘附和在流动负载下的高细胞保留。径向流动室安装有三区域特异性蛋白质固定基质（用于整合素的纤连蛋白，以及用于 VEGF 受体的 VEGF 和抗 Flk-1 抗体），可明显区分内皮细胞 (EC) 的剪切应力依赖性细胞保留潜力；这里使用EC作为接近EPC的细胞模型，因为EPC难以收获），这取决于固定蛋白的类型和细胞粘附期。高剪切应力抗性潜力的顺序为纤连蛋白> VEGF和抗Flk-1抗体。细胞粘附区域的粘附电位和细胞形状均通过相差显微镜和扫描电子显微镜图像确定。使用全内反射荧光显微镜测定荧光免疫染色的纽蛋白（粘着斑的关键蛋白）的荧光强度的分布和强度。所有这些都表明圆形细胞比分散良好的形状细胞更容易分离。本文讨论了一种潜在的原位 EPC 收获技术。