Use of the acyl azide method for cross‐linking collagen‐rich tissues such as pericardium

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Abstract
Collagen biomaterials should be cross‐linked in order to prevent biodegradation when they are used as implants. We have compared the cross‐linking efficiencies of glutaraldehyde and acyl azide in pericardium. Glutaraldehyde is used currently, but it elicits a cytotoxic effect which reduces the biocompatibility of cross‐linked tissue. We have attempted to overcome this problem by developing a cross‐linking method that obviates incorporation of foreign agents. Our process involves transformation of free carboxyl groups on collagen into acyl azide groups, which react with free amino groups on adjacent side chains. We have shown that the greatest increase in the thermal stability of collagen, as measured by differential scanning calorimetry, is achieved when tissue swelling is inhibited by the addition of sodium chloride (1 M) during acyl azide formation. Under these conditions, the denaturation temperature (Td) of pericardial collagen treated with acyl azide is raised to 83.4°C and that of tissue treated with glutaraldehyde to 85.1°C. Moreover, acyl‐azide‐treated tissues have the same resistance as glutaraldehydetreated tissues to chemical solubilization by cyanogen bromide and to enzymatic digestion by collagenase.