Hemocompatibility of treated polystyrene substrates: Contact activation, platelet adhesion, and procoagulant activity of adherent platelets

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Abstract
Platelet adhesion to biomaterials is often used as an index of blood compatibility, but a more clinically relevant issue is whether the adherent platelets are able to promote clot formation (i.e., if they are in the procoagulant state). Platelets rapidly generate thrombin when they are in the procoagulant state and the VA/Xa complex is present. We found that adherent platelets are procoagulant by three different methods: binding of FITC-Annexin V, acceleration of thrombin generation in the presence of Xa, Va, and prothrombin; and clotting of recalcified plasma. In the clotting times studies, the effect of adherent platelets on TCPS was completely eliminated by the addition of Annexin V, which is known to bind tightly to procoagulant platelets. The degree of procoagulant activity of adherent platelets was determined by measuring thrombin generation rates in the presence of the clotting factors Va, Xa, and prothrombin and normalizing to the number of adherent platelets. Two key observations were made in these studies. First, the procoagulant activity of platelets adherent to untreated and to several types of treated polystyrenes, as well as to glass and PET, was much greater than the procoagulant activity of unstimulated bulk phase platelets. Little difference in the procoagulant activity of adherent platelets was observed among the materials tested, however. Second, the procoagulant activity of platelets prestimulated with ionophore and subsequently allowed to adhere to Plastek M was much greater than when adherent platelets were stimulated by the adhesion event only. Measured values for platelet adhesion, platelet activation, and contact activation of blood plasma are discussed in the context of their potential combined impact on blood clotting.