Effect of applied uniaxial stress on rate and mechanical effects of cross-linking in tissue-derived biomaterials

Abstract

Conformational changes in collagen fibrils, and indeed the triple helix, can be produced by application of mechanical stress or strain. We have demonstrated that the rate of cross-linking in glutaraldehyde and epoxide homobifunctional reagents can be modulated by uniaxial stress (strain). Two poly(glycidyl ether) epoxides were used: Denacol ® EX-810 (a small bifunctional reagent), and Denacol EX-512 (a large polyfunctional reagent). To prevent any possible effect from being masked by saturation of cross-linking sites, bovine pericardium was cross-linked to such an extent that the increase in collagen denaturation temperature, T d , was one-half of the maximal rise achievable with each reagent. Uniaxial tensile stress of 0, 15, 124 or 233 kPa was applied during cross-linking. Cross-linking rate (as observed by increase in T d ) increased with increasing stress to a maximum at 124 kPa in glutaraldehyde at pH 7 but decreased in EX-810 at pH 7. In each case, the effect was small but statistically significant. No effect was observed with the larger EX-512. Cross-linking under increasing stress also showed systematic effects on mechanical properties: decreasing extensibility and plastic strain while increasing tensile strength. In each case, the effects of the epoxides were slightly different from those of glutaraldehyde. In preparation for the above experiments, studies of the effect of pH, temperature, and exposure time were carried out for each epoxide and (to a lesser extent) for glutaraldehyde. Again, systematic changes in mechanical properties were observed with increasing T d . Conformational changes in collagen produced by mechanical stress (strain) modulate the rate of cross-linking and the resulting mechanical properties; however, the effects are sensitive to the reagent employed.