The in vitro and in vivo behavior of urokinase immobilized onto collagen-synthetic polymer composite material.

With a view to developing biomaterials for semipermanent substitution, we have studied a composite material constituted with collagen and a synthetic polymer which possesses high tissue compatibility. This collagen-synthetic polymer composite was applied as a support for immobilization of enzymes for the purpose of providing a material surface with biological function. The enzymes, urokinase and trypsin, were successfully bound to the collagen membrane layer which had been activated by acyl azide formation of its carboxyl groups. The enzyme-bearing composite material showed excellent catalytic activity toward a protein substrate as well as a low-molecular-weight synthetic substrate. The immobilized urokinase was characterized enzymatically and compared with native urokinase. The apparent affinity of immobilized urokinase for the substrate was slightly decreased, but its intrinsic kinetic properties were not significantly affected. No decrease in its esterase activity was observed both on repeated use and on long-term storage, and its fibrinolytic activity was stable on heat or disinfection treatment. When this urokinase-bearing composite material was applied into rabbit blood vessels, its in vivo fibrinolytic activity was maintained. Thus, enzyme-collagen-synthetic polymer composites may find wide application for biomaterials and artificial organs as functional biomaterials.