Polymer brushes prepared via surface-initiated polymerization of 2-hydroxyethyl methacrylate are powerful platforms for the fabrication of model biointerfaces to study cell-substrate interactions. In this manuscript, the versatility of surface-initiated polymerization and the poly(2-hydroxyethyl methacrylate) (PHEMA) polymer brush platform are used to address two fundamental questions, viz. the effects of ligand co-presentation and of the 3D localization of biochemical cues on cell behavior. Using a series of PHEMA brushes that present RGD and PHSRN ligands in various relative surface concentrations, the present study unequivocally demonstrates that: (i) co-presentation of PHSRN cues on an RGD functionalized substrate enhances cell adhesion and (ii) this synergetic effect is highest when the two ligands are presented at equal surface concentrations. In the second part of this study, adhesion of 3T3 fibroblasts on a series of PHEMA brushes that present the RGD ligand at a distance of 12, 23 or 42 nm away from the cell substrate interface is investigated. While cells were found to adhere to surfaces that presented the cell adhesive peptides at distances up to 23 nm from the interface, polymer brushes that contained the RGD ligands 42 nm away from the interface did not support cell adhesion.
Keywords: Cell adhesion; Fibroblast; PolyHEMA; RGD peptide; Surface grafting.
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