A Biologically Active Surface Enzyme Assembly that Attenuates Thrombus Formation

Abstract

Activation of hemostatic pathways by blood‐contacting materials remains a major hurdle in the development of clinically durable artificial organs and implantable devices. Here, it is postulated that surface‐induced thrombosis may be attenuated by the reconstitution onto blood contacting surfaces of bioactive enzymes that regulate the production of thrombin, a central mediator of both coagulation and platelet activation cascades. Thrombomodulin (TM), a transmembrane protein expressed by endothelial cells, is an established negative regulator of thrombin generation in the circulatory system. Traditional techniques to covalently immobilize enzymes on solid supports may modify residues contained within or near the catalytic site, thus reducing the bioactivity of surface enzyme assemblies. In this report, a molecular engineering and bioorthogonal chemistry approach to site‐specifically immobilize a biologically active recombinant human TM fragment onto the luminal surface of small diameter prosthetic vascular grafts is presented. Bioactivity and biostability of TM modified grafts is confirmed in vitro and the capacity of modified grafts to reduce platelet activation is demonstrated using a non‐human primate model. These studies indicate that molecularly engineered interfaces that display TM actively limit surface‐induced thrombus formation. A biologically active thrombomodulin surface assembly to limit interfacial thrombin production was generated on the lumen of clinical ePTFE vascular grafts by site‐specific covalent immobilization. Therapeutic capacity of this biomimetic surface engineering approach to attenuate thrombus formation was demonstrated in a clinically relevant in vivo model of prosthetic graft thrombosis.