High fibrinogen γ′ levels in patient plasma increase clot formation at arterial and venous shear

Abstract

Abstract Fibrinogen γ' accounts for 3% to 40% of plasma fibrinogen. Earlier studies indicated that fibrinogen γ' forms altered fibrin clots under static conditions, whereas clinically, altered plasma γ' levels are associated with arterial and venous thrombosis. However, the effects of static vs flow conditions on the role of γ′ throughout the pathophysiological range is unknown. This study explores the effects of γ' levels on clot formation and structure in static and flow conditions. Coagulation of plasma samples with low (n = 41; 3%), normal (n = 45; 10%), or high (n = 33; 30%) γ′ levels were compared with that of purified fibrinogen mixtures with increasing ratios of γ′ (3%, 10%, 30%). Clots were analyzed by confocal microscopy, permeation, turbidity, and lysis techniques. In a novel 2-step flow-perfusion model, fibrinogen-deficient plasma repleted with increasing ratios of γ′ (3%, 10%, 30%) or plasmas with low (n = 5, 3%) or high (n = 5, 30%) γ′ were flowed over preformed platelet aggregates at arterial (500 s−1) and venous (150 s−1) shear rates. Increasing γ′ percentages within the pathophysiological range (3%-30%) did not result in any change in clot-formation rates; however, it led to significantly higher clot density, thinner fibers, and slower lysis in static conditions. Under flow at arterial shear, high γ′ (30%) led to faster (+44.1%-75.3%) and increased (+104%-123%) fibrin deposition, with clots exhibiting a larger volume (+253%-655%) and height (+130%-146%). These trends were magnified at venous shear. Overall, our findings demonstrate the significant impact of pathophysiological fibrinogen γ′ levels on clot structure and provide new flow-dependent mechanisms to explain how γ′ increases thrombosis risk.