Carbodiimide cross-linking counteracts the detrimental effects of gamma irradiation on the physical properties of collagen-hyaluronan sponges

Collagen-based scaffolds are extensively used in biomaterials and tissue engineering applications. These scaffolds have shown great biocompatibility and versatility, but their relatively low mechanical properties may limit use in orthopaedic load-bearing applications. Moreover, terminal sterilization with gamma irradiation, as is commonly performed with commercial devices, presents concerns over structural integrity and enzymatic stability. Therefore, the goal of this study was to test the hypothesis that EDC/NHS cross-linking (10 mM/5 mM) can protect collagen-hyaluronan sponges from the damaging effects of gamma irradiation. Specifically, we evaluated compressive and tensile mechanical properties, enzymatic stability, porosity and pore size, and swelling ratio. Ultimate tensile strength and elastic modulus exhibited increases (168.5 and 245.8%, respectively) following irradiation, and exhibited over tenfold increases (1049.2 and 1270.6%, respectively) following cross-linking. Irradiation affected pore size (38.4% decrease), but cross-linking prior to irradiation resulted in only a 17.8% decrease. Cross-linking also showed an offsetting effect on the equilibrium modulus, enzymatic stability, and swelling ratio of sponges. These results suggest that carbodiimide cross-linking of collagen-hyaluronan sponges can mitigate the structural damage typically experienced during gamma irradiation, warranting their use in tissue engineering applications.