title: A quantitative study of MC3T3-E1 cell adhesion, morphology and biomechanics on chitosan-collagen blend films at single cell level.
abstract: The interaction between cells and biomaterials plays a key role in cell proliferation and differentiation in tissue engineering. However, a quantitative analysis of those interactions has been less well studied. The objective of this study was to quantitative recapitulate the difference of MC3T3-E1 cell adhesion, morphological and biomechanical properties on chitosan-collagen films in terms of chemical composition. Here, the unbinding force between MC3T3-E1 cell and a series of chitosan-collagen films was probed by a real-time and in situ atomic force microscopy-single cell force spectroscopy (AFM-SCFS). Meanwhile, changes in cell morphology and Young's modulus on different chitosan-collagen films were detected by AFM. The cell area and CCK-8 results showed that cell spreading and proliferation increased with increasing collagen content. AFM observations clearly showed cell height decreased and pseudopod fusion with the collagen content increased. Cell adhesive force increased from 0.76±0.17 nN to 1.70±0.19 nN. On the contrary, cells Young's modulus, which reflected biophysical changes of cells decreased from 11.94±3.19 kPa to 1.81±0.52 kPa, respectively. It suggested that stronger cell-substrate interactions benefit cell adhesion, and better cell flexibility improve cell spreading. The findings indicate that cell morphology, adhesive force and Young's modulus are significant affected by various chitosan-collagen substrates. Those methods and quantitative results have guiding significance for investigating the mechanism of chitosan and/or collagen based cell-targeting drug carrier and the preparation of chitosan-collagen composite biomaterials.
authors.affiliation: Department of Materials Science and Engineering, Jinan University, Guangzhou 510632, PR China; Engineering Research Center of Artificial Organs and Materials, Ministry of Education, Guangzhou 510632, PR China.
authors.forename: Chuang
authors.initials: C
authors.lastname: Wang
authors.affiliation: Department of Materials Science and Engineering, Jinan University, Guangzhou 510632, PR China; Engineering Research Center of Artificial Organs and Materials, Ministry of Education, Guangzhou 510632, PR China.
authors.forename: Xu-dong
authors.initials: XD
authors.lastname: Xie
authors.affiliation: Department of Materials Science and Engineering, Jinan University, Guangzhou 510632, PR China; Engineering Research Center of Artificial Organs and Materials, Ministry of Education, Guangzhou 510632, PR China.
authors.forename: Xun
authors.initials: X
authors.lastname: Huang
authors.affiliation: Experiment and Technology Center, Jinan University, Guangzhou 510632, PR China. Electronic address: zhihong3615@sina.com.
authors.forename: Zhi-hong
authors.initials: ZH
authors.lastname: Liang
authors.affiliation: Department of Materials Science and Engineering, Jinan University, Guangzhou 510632, PR China; Engineering Research Center of Artificial Organs and Materials, Ministry of Education, Guangzhou 510632, PR China. Electronic address: tcrz9@jnu.edu.cn.
authors.forename: Chang-ren
authors.initials: CR
authors.lastname: Zhou
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001696:Biomechanical Phenomena; D002448:Cell Adhesion; D048271:Chitosan / Q000737:chemistry*; D003094:Collagen / Q000737:chemistry*; D051379:Mice; D018625:Microscopy, Atomic Force; D059010:Single-Cell Analysis
chemical_list: D003094:Collagen; D048271:Chitosan
keywords: Atomic force microscopy; Chitosan; Collagen; Single cell force spectroscopy; Young's modulus