Assessment of material-induced procoagulant activity by a modified Russell viper venom coagulation time test

Platelet activation is an inevitable consequence of blood-material interactions. The ability of those activated platelets and platelet-derived microparticles to enhance coagulation reactions leading to thrombin/fibrin formation has not been well studied despite its potential significance. Activated platelets and platelet-derived microparticles are known to dramatically enhance the catalytic efficiencies of the tenase and prothrombinase complexes. In this paper, a modified Russell viper venom coagulation time test is used to quantitate material-induced procoagulant activity due to the generation of activated phospholipid surfaces. In our test system, polyethylene and Silastic tubes were filled with heparinized whole blood and left to gently flow back and forth at 37 degrees C. After 1 h, the blood within the tubes was gravity drained and the plasma fraction assayed for procoagulant activity. The clotting times were determined by a Coag-A-Mate X2 instrument after the automated addition of Russell viper venom (to activate factors V and X) and calcium ions. Appreciable procoagulant activity was generated during whole blood contact within polyethylene and Silastic tubes although significantly greater activity was generated by the latter surface. As previously reported, platelet-derived microparticles also were detected by flow cytometry. Filtration of the plasma after material contact through a 0.1 microm filter led to substantial gains in clotting times and to near complete removal of microparticles, indicating that the material-induced microparticles likely were responsible for the procoagulant activity.