Fibronectin adsorption and arrangement on copolymer surfaces and their significance in cell adhesion
Fibronectin adsorption and arrangement on copolymer surfaces and their significance in cell adhesion
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DOI：
10.1002/jbm.a.30238
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发表时间：
2005-02-01
影响因子：
4.9
通讯作者：
Kaminski, J
中科院分区：
文献类型：
--
作者：
Kowalczynska, HM;Nowak-Wyrzykowska, M;Kaminski, J
纤维连接蛋白（FN）的吸附（苯乙烯/甲基丙烯酸甲酯）共聚物表面，磺化（亲水性）和norisulfonated（疏水性），研究了通过放射性同位素（I-125-FN）和ELISA测定;后者采用单克隆抗体。结果发现，放射性碘衍生的等温线不遵循朗缪尔型吸附法的FN浓度范围内的研究，而是，观察到一个准线性FN表面密度与体积浓度的依赖性。这些等温线，和我们最近的ELISA测量与多克隆抗体，使我们能够估计饱和FN表面密度，这是在实验误差内，这两种类型的表面相似。这表明吸附的FN的量与观察到的白血病L1210细胞粘附的差异无关（FN涂覆的磺化表面比其非磺化类似物更具亲粘附性）。通过使用针对不同FN结构域的单克隆抗体，以及通过用合成的Gly-Arg-Gly-Asp-Ser-Pro（GRGDSP）肽阻断α（5）β（1）整联蛋白受体，证实了这些差异是由FN排列变化诱导的假设。位于FN细胞结合结构域内的RGD序列似乎在非磺化表面上采用的结构中被掩蔽，这阻碍了整合素-配体相互作用。(C)2004 Wiley Periodicals，Inc.
The adsorption of fibronectin (FN) to (styrene/methyl methacrylate) copolymer surfaces, both sulfonated (hydrophilic) and norisulfonated (hydrophobic), was studied by means of the radioisotope (I-125-FN) and ELISA assays; the latter employed monoclonal antibodies. It was found that the radioiodination-derived isotherms did not follow the Langmuir-type adsorption law within the FN concentration range studied; rather, a quasi-linear FN surface density versus bulk concentration dependence was observed. These isotherms, and our recent ELISA measurements with polyclonal antibodies, allowed us to estimate saturative FN surface densities, which were, within the experimental error, similar on both types of surfaces. This suggested the amount of adsorbed FN to be not responsible for observed differences in leukaemia L1210 cell adhesion (FN-coated sulfonated surfaces are far more pro-adhesive than their nonsulfonated analogues). The presumption that these differences are induced by changes in the FN arrangement was confirmed by the use of monoclonal antibodies directed against distinct FN domains, and by the blocking of alpha(5)beta(1) integrin receptor with the synthetic Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP) peptide. The RGD sequence located within the FN cell-binding domain seems to be masked in the structure adopted on nonsulfonated surfaces, which hinders the integrin-ligand interaction. (C) 2004 Wiley Periodicals, Inc.