Abstract
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.
Highlights
Natural polymers play an important role in biomaterials due to their good biocompatibility and biodegradability
Analysis of biological pathways The results of biological pathway analysis indicate 40 biological pathways involved in the protein adsorption on the chitosan surface and 19 on the collagen/chitosan surface; 16 pathways were activated on both surfaces (Supplementary Table S4)
The collagen/chitosan film was more conducive to promoting cell adhesion and growth
Summary
Natural polymers play an important role in biomaterials due to their good biocompatibility and biodegradability. Chitosan and collagen are two of the most widely-used natural polymeric materials in the biomaterials. Due to its excellent biocompatibility, biodegradability, physiological activity and film-forming property, chitosan has become a widespread biomaterial in the biomedical field [1]. Some studies showed that a pure chitosan film have a certain inhibitory effect on cell adhesion and proliferation. The reason is that the presence of primary amine of glucosamine residue makes chitosan as a pH-responsive polycation; a small change in environmental pH can affect protein adsorption [2], thereby affecting cell adhesion. A large number of cell biocompatibility studies on chitosan have been reported and are mainly performed on film, including pure chitosan film and chitosan blends such as collagen/chitosan film [3, 4]
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