Effects of Substratum Topography on Vascular Endothelium

描述（由申请人提供）：基底膜具有许多特征，这些特征极大地影响细胞功能，包括特定蛋白质和官能团、生长因子和其他营养剂的储存器以及粘附细胞延伸突起进入其中并形成粘附斑的复杂三维形貌。底层基质的三维形貌，独立于特定的受体-配体相互作用，最近已被证明会影响基本的细胞行为。迄今为止进行的大多数研究已经评估了大尺度（>1微米）特征对细胞行为的影响。这些“大规模”研究与体内细胞行为的相关性尚不清楚，因为我们的实验室已经表明，非血管组织中的基底膜由复杂的三维纳米级（1微米特征尺寸）结构组成，该结构显着放大了其细胞-膜相互作用的表面积。该提案的总体目的是确定血管内皮下的基底膜的地形特征，并研究基底上存在的纳米级（< 1微米）地形特征的细胞后果。在本申请中，提出了一种多学科的方法来测试3个假设，使用定量形态学技术，在细胞生物学，分子生物学，和国家的最先进的纳米制造技术的体外方法。假设1：血管内皮基底膜的地形特征在血管类型和位置之间是相似的，并且与全身其他基底膜的地形特征相似。假设二：完全合成的表面可以通过控制制造来工程化，具有生物相关的特征类型、尺寸和分布，这将调节血管内皮行为。假设3：基底膜的地形特征提供细胞外线索，导致通过参与粘着斑复合物的蛋白质进行信号传导。

DESCRIPTION (provided by applicant): Basement membranes have many features, which greatly influence cell function including specific proteins and functional groups, a reservior of growth factor and other trophic agents, and a complex three dimensional topography into which adherent cells extend processes, and to which cells form adhesion plaques. The three-dimensional topography of the underlying substrate, independent of specific receptor-ligand interactions, has been recently shown to influence fundamental cell behaviors. The majority of studies conducted to date have evaluated the effect of large scale (>1 micron) features on cell behavior. The relevance of these "large-scale" studies to cell behavior in vivo is not clear since our laboratories have shown basement membranes in non-vascular tissues to consist of a complex 3-dimensional nanoscale (1 micron feature size) architecture which dramatically amplifies its surface area for cell-membrane interaction. The overall purpose of this proposal is to to determine the topographic features of the basement membrane underlying the vascular endothelium and to investigate the cellular consequences of nanoscale (< 1 micron) topographic features present on the substratum. In this application, a multi-disciplinary approach is proposed to test 3 hypotheses using quantitative morphologic techniques, in vitro methodologies in cell biology, molecular biology, and state-of-the-art nanoscale fabrication techniques. Hypothesis 1: The topographic features of vascular endothelial basement membranes is similar between vessel types and locations and similar to that found for other basement membranes throughout the body. Hypothesis 2: Totally synthetic surfaces can be engineered through controlled fabrication with biologically relevant feature types, dimensions and distributions that will modulate vascular endothelial behaviors. Hypothesis 3:The topographic features of the basement membrane provide extracellular cues leading to signaling through proteins involved in focal adhesion complexes.