The Role of Fibrinogen-Mediated Platelet Aggregation in Subsequent Platelet-Driven Blood Clot Contraction

Abstract

BackgroundBlood clot contraction/retraction depends on the force-generating actomyosin and on the platelet integrin IIb{beta}3, which transmits intracellular forces to fibrin. Before clotting, fibrinogen binds to activated integrin IIb{beta}3, mediating platelet aggregation. The relationship between platelet aggregation and subsequent platelet-driven clot contraction remains unclear. MethodsWe investigated the effects of platelet aggregation on clot contraction by selectively blocking the IIb{beta}3-fibrinogen binding using the RGDW peptide. The ability of RGDW to disrupt IIb{beta}3-fibrinogen binding was assessed by platelet aggregometry. The time-course of clot contraction was monitored optically in whole blood or platelet-rich plasma and modeled mathematically. Clot stiffness was assessed using Thromboelastography. The effect of the RGDW peptide on the structure of PRP-clots was examined using scanning electron microscopy. ResultsThe RGDW peptide dose-dependently inhibited TRAP-induced platelet aggregation. Both in whole blood and in plasma, the peptide dose-dependently prolonged the lag-period and slowed the rate without affecting the final extent of contraction. Thromboelastography showed that RGDW dose-dependently increased maximum clot stiffness in blood. Scanning electron microscopy revealed that RGDW treatment resulted in formation of smaller fibrin agglomerates surrounding non-aggregated platelets. A theoretical model allowed us to decipher mechanisms underlying the kinetic effects of RGDW. ConclusionBlocking the binding of integrin IIb{beta}3 to fibrinogen and preventing platelet aggregation delays and slows subsequent clot contraction without affecting the final degree of shrinkage. These findings indicate a modulatory role of fibrinogen-mediated platelet aggregation in clot contraction and highlight the unforeseen effects of selective inhibitors of platelet aggregation on the contraction of blood clots and thrombi.