Background: The functional links between the activation of platelets and the coagulation system have not been clarified.
Methods and results: Immobilized collagen fibrils were perfused with human blood containing fluoresceinated platelets in the presence of various concentrations of thrombin inhibitor. Coagulant activity around platelet thrombi was detected using a FITC-conjugated antibody against the fibrin monomer complex (F-405). Intra-cytosolic calcium ion concentrations ([Ca(2+)](i)) in individual platelets and the volume of thrombi were detected with an ultrafast confocal laser microscope equipped with a piezo-motor control unit. The volume of platelet thrombi formed after 8 min of blood perfusion in the presence of 10, 25, 50, and 100 micromol/L argatroban was 7.69+/-0.46 microm(3), 6.61+/-1.96 microm(3), 3.63+/-1.54 microm(3), and 1.67+/-0.75 microm(3), respectively. There was a positive correlation between the volume of platelet thrombi and the amount of fibrin monomer complex produced around them. The [Ca(2+)](i) of the platelets forming the thrombi oscillated between a minimum of 92.0+/-57.4 nmol/L, 120.1+/-68.1 nmol/L, and a maximum of 217.6+/-131.5 nmol/L, 367.6+/-189.1 nmol/L, respectively, in the presence of 100 and 10 mumol/L argatroban.
Conclusions: The results suggest a crucial role of coagulant activity in both the generation of fibrin and the growth of platelet thrombi.