Lü Xiaoying, Zhang Heng, Huang Yan, Zhang Yiwen
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, P.R. China.
SQ Medical Device Co., Ltd., Nanjing, P.R. China.
Regen Biomater. 2018 Oct;5(5):261-273. doi: 10.1093/rb/rby017. Epub 2018 Jul 17.
The aim of this article is to apply proteomics in the comparison of the molecular mechanisms of PC12 cell adhesion and growth mediated by the adsorbed serum proteins on the surfaces of chitosan and collagen/chitosan films. First, the chitosan and the collagen/chitosan films were prepared by spin coating; and their surface morphologies were characterized by scanning electron microscopy, X-ray energy dispersive spectroscopy, contact angle measurement and Fourier transform infrared spectroscopy. Subsequently, cell proliferation experiments on two materials were performed and the dynamic curves of protein adsorption on their surfaces were measured. Then, proteomics and bioinformatics were used to analyze and compare the adsorbed serum proteins on the surfaces of two biomaterials; and their effects on cell adhesion were discussed. The results showed that the optimum concentration of chitosan film was 2% w/v. When compared with chitosan film, collagen/chitosan film promoted the growth and proliferation of PC12 cells more significantly. Although the dynamic curves showed no significant difference in the total amount of the adsorbed proteins on both surfaces, proteomics and bioinformatics analyses revealed a difference in protein types: the chitosan surface adsorbed more vitronectin whereas collagen/chitosan surface adsorbed more fibronectin 1 and contained more cell surface receptor binding sites and more Leu-Asp-Val sequences in its surface structure; the collagen/chitosan surface were more conducive to promoting cell adhesion and growth.
本文旨在应用蛋白质组学比较壳聚糖以及胶原/壳聚糖膜表面吸附的血清蛋白介导PC12细胞黏附与生长的分子机制。首先，通过旋涂法制备壳聚糖和胶原/壳聚糖膜；并通过扫描电子显微镜、X射线能谱、接触角测量以及傅里叶变换红外光谱对其表面形态进行表征。随后，对两种材料进行细胞增殖实验，并测定其表面蛋白质吸附的动态曲线。然后，运用蛋白质组学和生物信息学分析并比较两种生物材料表面吸附的血清蛋白；并探讨它们对细胞黏附的影响。结果表明，壳聚糖膜的最佳浓度为2%（w/v）。与壳聚糖膜相比，胶原/壳聚糖膜更显著地促进了PC12细胞的生长和增殖。尽管动态曲线显示两种表面吸附蛋白的总量无显著差异，但蛋白质组学和生物信息学分析揭示了蛋白质类型的差异：壳聚糖表面吸附了更多的玻连蛋白，而胶原/壳聚糖表面吸附了更多的纤连蛋白1，并且其表面结构含有更多的细胞表面受体结合位点和更多的亮氨酸-天冬氨酸-缬氨酸序列；胶原/壳聚糖表面更有利于促进细胞黏附与生长。
