A Novel Surface Modification Using Tissue Factor Reconstituted in Phospholipid Vesicles for the Activation of Blood Coagulation

Abstract

We describe a novel method to immobilize recombinant human tissue factor (rhTF) reconstituted in phospholipid vesicles. The rhTF vesicles were immobilized in a multilayer vesicle structure using cholesterol–DNA tethers spontaneously inserted into the lipid membrane. The properties of the rhTF vesicle surface modification were characterized by surface plasmon resonance biosensor technology. As an application of this surface modification, we investigated its use as a blood coagulation activating surface. The coagulation activating capacity of the surface modification was tested by exposure to human whole blood in a flow chamber. No increase in rhTF levels in the blood was found after passage through the flow chamber, indicating that the rhTF surface modification was stable. Thrombin–antithrombin (TAT) and prothrombin fragment (PF) 1 + 2 levels increased after exposure to the surface, and decreased in a concentration-dependent way upon addition of melagatran (a direct thrombin inhibitor), i.e., coagulation activity triggered by rhTF could be suppressed by anticoagulation. The results with this new thrombogenic surface are promising, and will be further developed into a useful tool for coagulation related investigations, e.g., characterization of anticoagulants and biomaterials.

Keywords:

• TISSUE FACTOR
• TETHERED LIPID VESICLES
• CHOLESTEROL-DNA
• BLOOD COAGULATION
• FLOW CHAMBER