The properties of collagen films crosslinked by physical and chemical techniques were compared to the properties of films crosslinked with glutaraldehyde (GTA). Physical techniques studied include exposure to short wave (254 nm) u.v. irradiation and severe dehydration. Chemical techniques studied include immersion of collagen films in aqueous solutions of cyanamide or GTA. Collagen films exposed to combinations of aqueous solutions of cyanamide and severe dehydration had moduli of elasticity, swelling ratios and resistance to bacterial collagenase similar to films crosslinked with GTA. Theoretical calculations based on amino acid composition indicate that approximately seven times as many amino acid residues are capable of forming crosslinks using cyanamide or severe dehydration procedures as compared to GTA crosslinking. In addition, using severe dehydration or cyanamide forms crosslinks involving both amino and carboxyl residues which may allow these procedures to act synergistically. Based on our studies this two-step procedure effectively crosslinks collagen-based biomaterials while the only by-product of this reaction is water-soluble urea. Preliminary biocompatibility studies suggest that this crosslinking procedure may allow for pronounced tissue ingrowth.

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