Adsorbed layers of oriented fibronectin: A strategy to control cell-surface interactions
Adsorbed layers of oriented fibronectin: A strategy to control cell-surface interactions
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DOI：
10.1002/jbm.a.30417
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发表时间：
2005-11-01
影响因子：
4.9
通讯作者：
Van Tassel, PR
中科院分区：
文献类型：
--
作者：
Calonder, C;Matthew, HWT;Van Tassel, PR
纤维连接蛋白(FN)是一种基质蛋白，可通过其细胞结合部位和相关的协同作用部位诱导细胞附着和扩散。因此，FN涂层表面在组织工程和其他细胞接触应用中很有用，但许多固定策略的问题是分子取向的随机分布。我们试图通过羧甲基葡聚糖层固定FN来控制FN的取向，从而提高其细胞结合部位的可用性，在该层上化学地附着了针对FN C末端附近区域(从而远离细胞结合部位)的单抗。使用光波导光模式光谱，我们显示了化学偶联抗体的存在，以产生相当致密和较厚的FN层，与更垂直排列的蛋白质一致。与对照FN层(即没有结合抗体)相比，人脐静脉内皮细胞在定向FN层(即在存在固定化单抗的情况下)上的扩散速度显著加快，并且以更球状对称的方式传播。然而，我们观察到人脐静脉内皮细胞在定位的FN层上的扩散类似于没有羧甲基葡聚糖层的FN层，这表明尽管定位FN是一个有前途的策略，但可能需要使用除葡聚糖之外的连接子的偶联策略。(C)2005年威利期刊公司。
Fibronectin (Fn) is a matrix protein known to induce cell attachment and spreading through its cell binding site and related synergy sites. Fn-coated surfaces are therefore useful in tissue engineering and other cell contacting applications, but a problem with many immobilization strategies is a random distribution of molecular orientations. We sought to control Fn orientation, and thus enhance the availability of its cell binding site, by immobilizing Fn via a carboxymethyl dextran layer onto which are chemically attached monoclonal antibodies specific to a region near to Fn's C terminus (and thus away from the cell binding site). Using optical waveguide lightmode spectroscopy, we show the presence of chemically coupled antibodies to yield a considerably denser and thicker Fn layer, consistent with a more vertically aligned protein. Human umbilical vein endothelial cells spread significantly faster, and in a more spherically symmetric way, on an oriented Fn layer (i.e., in the presence of immobilized monoclonal antibodies) as compared with a control Fn layer (i.e., in the absence of bound antibodies). However, we observe human umbilical vein endothelial cell spreading on the oriented Fn layer to be similar to that on a Fn layer in the absence of a carboxymethyl dextran layer, suggesting that although orienting Fn is a promising strategy, coupling strategies using linkers other than dextran may be needed. (c) 2005 Wiley Periodicals, Inc.