Adhesion and proliferation properties of type I collagen-derived peptide for possible use in skin tissue engineering...

The surface properties of three-dimensional scaffolds are improved by coating or covalently linking certain adhesion-promoting proteins or peptides. In the present study, the effect of type I collagen-derived peptide (GKNGDDGEA) on adhesion and proliferation of HaCaT keratinocytes and NIH3T3 murine fibroblast cell lines was studied to assess its suitability for possible skin tissue engineering applications. Cell adhesion and proliferation of HaCaT and NIH3T3 were found to be enhanced by peptide coating. The optimum peptide coating densities to obtain the best cell adhesion and proliferation were found to be 0.827 µmoles/cm² and 0.62 µmoles/cm² for HaCaT and NIH3T3, respectively. Cell adhesion, in the presence of anti-integrin α1 antibody, inhibited attachment of NIH3T3 cells indicating the involvement of integrin α1 receptor. However, the attachment of HaCaT cells was not affected by anti-integrin treatment. The higher expression of paxillin confirmed the effect of the peptide in mediating focal adhesion kinases (FAKs) in cell adhesion and proliferation. Gene expression analysis was performed on cell migration proteins like Rho, Rac, Cdc42, integrin receptor α1, and β1, and the extracellular matrix modulating proteins like MMP2, TIMP, and COL1A1 to validate their role on the peptide-mediated cell proliferation. Immunofluorescence analysis showed the distribution and localisation of phospho-FAK on cells cultured on the peptide-coated surfaces. Results support the role of peptides in enhancing cell adhesion and proliferation properties.