Fibronectin distribution on demixed nanoscale topographies
Fibronectin distribution on demixed nanoscale topographies
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DOI：
10.5301/ijao.2011.6316
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发表时间：
2011-01-01
影响因子：
1.7
通讯作者：
Salmeron-Sanchez, Manuel
中科院分区：
文献类型：
--
作者：
Perez-Garnes, Manuel;Gonzalez-Garcia, Cristina;Salmeron-Sanchez, Manuel
目的：已知表面纳米形貌影响细胞的黏附和分化。我们的目的是通过研究纤维连接蛋白在材料界面的吸附中间层的状态，分析纳米形貌对纤维连接蛋白吸附的影响，进而影响细胞与材料的黏附，从而使细胞与材料的相互作用更加合理。方法：通过高速旋转浇注过程中薄膜聚合物共混物PLLA和PS的分离，形成纳米形貌表面。纤维连接蛋白(FN)被吸附在不同的纳米形貌上，并通过原子力显微镜(AFM)直接观察到蛋白质的分布。通过图像分析，定量了蛋白质覆盖表面的比例，以及Fn在峰和谷之间的分布。结果：在3~30 nm的高度范围内，通过改变体系的组成得到了不同的纳米尺度结构域。FN倾向于吸附在纳米岛状结构的峰上，特别是在没有增强细胞黏附的组合物中。此外，蛋白质在谷峰之间的分布改变了焦点黏附斑块的大小，在纤维连接蛋白均匀分布的表面，黏附斑块变大。结论：表面纳米形貌是影响蛋白质吸附的关键材料特性。此外，根据斑块的大小，蛋白质在不同样本上的分布与细胞最初的黏附能力有关。
Purpose: It is known that surface nanotopography influences cell adhesion and differentiation. Our aim is to analyze the effect of nanoscale topography on fibronectin adsorption and, afterwards, on cell adhesion in order to rationalize the cell-material interaction by focusing on the state of the intermediate layer of adsorbed fibronectin at the material interphase.Methods: Nanotopographic surfaces were produced by demixing of thin film polymer blends - PLLA and PS - during a high speed spin-casting process. Fibronectin (FN) was adsorbed on the different nanotopographies and the protein distribution was directly observed by atomic force microscopy (AFM). The fraction of the surface covered by the protein was quantified by image analysis, as well as the distribution of FN between peaks and valleys. Focal adhesion protein -vinculin- was immunostained and quantified by image analysis on the different nanoscale surfaces.Results: Different nanoscale domains were obtained by changing the composition of the system within a height range of 3 nm to 30 nm. FN tends to adsorb on the peaks of nanoisland topographies, especially in compositions that did not enhance cell adhesion. Moreover, protein distribution between valleys and peaks alters the size of focal adhesion plaques, which grew larger on surfaces with an even distribution of fibronectin.Conclusions: Our results suggest that the surface nanotopography is a key material property capable of influencing protein adsorption. Additionally, the distribution of the protein on the different samples was correlated to the initial ability of cells to adhere in terms of the size of the focal plaques.