Incorporation of adhesion peptides into nonadhesive hydrogels useful for tissue resurfacing

Photopolymerized crosslinked networks of poly(ethylene glycol; PEG) diacrylate (MW 8000) were derivitized throughout their bulk with Arg-Gly-Asp (RGD)-containing peptide sequences. Incorporation was achieved by functionalizing the amine terminus of the peptide with an acrylate moiety, thereby enabling the adhesion peptide to copolymerize rapidly with the PEG diacrylate upon photoinitiation. PEG diacrylate hydrogels derivitized with RGD peptide at surface concentrations ranging from 0.001 to 1 pmol/cm2 were studied in vitro for their ability to promote spreading of human foreskin fibroblasts over 24 h. Hydrogels not derivitized with peptides were poor substrates for adhesion, permitting spreading of only 5% of the seeded cells. When immobilized with no spacer arm, both RGD and RDG (inactive control) supported spreading of approximately 50% and approximately 15% of cells at 1 and 0.1 pmol/cm2 surface concentrations respectively; lower concentrations did not promote spreading. When a MW 3400 PEG spacer arm was incorporated between the hydrogel and the peptide linkage, incorporation of 1 pmol/cm2 RGD promoted 70% spreading whereas RDG at the same concentration did not promote spreading. In addition, when cells were seeded in serum-free medium, only RGD peptides incorporated with a spacer arm were able to promote spreading. Thus peptide incorporated into PEG 8000 diacrylate hydrogels without a spacer arm nonspecifically mediated cell spreading whereas incorporation via a MW 3400 PEG spacer arm was required to permit cell spreading to be specifically mediated.