Fibrin clot formation under diverse clotting conditions: Comparing turbidimetry and thromboelastography

Abstract

Thrombosis is a leading cause of death around the world. Fibrin, the protein primarily responsible for clot formation, is formed via cleaving soluble fibrinogen by thrombin with resulting properties varying under different clot forming conditions. This study sought to compare trends across thromboelastography (TEG) and turbidimetry utilizing a simplified fibrinogen/thrombin clot model. Turbidimetry is an optical measure (550 nm) of fibrin clot formation and is widely utilized due to its laboratory accessibility and ease of use. Thromboelastography (TEG) is a specialized viscoelastic technique that directly measures clot strength and is primarily utilized in the clinical setting to assess patients' hemostasis. In these experiments, human and bovine fibrin clots were formed in-vitro by mixing fibrinogen and thrombin under diverse clotting conditions. Increasing thrombin concentration (0 to 10 U/mL), ionic strength (0.05 to 0.3 M), pH (5.5 to 8.1), and lowering albumin concentration (100 to 0 mg/mL) resulted in decreased clot turbidity and increased clot strength using species-matched bovine and human fibrinogen and thrombin. Whereas, increasing fibrinogen concentration (1 to 5 mg/mL) resulted in increased clot turbidity and increased clot strength for both species-matched and cross-species fibrinogen and thrombin. Clotting times with both techniques followed a similar trend and were observed to be unchanged when varying albumin concentration, elongated with increasing fibrinogen, and shortened with increasing pH, ionic strength, and thrombin. TEG and turbidimetry track clot formation via two distinct methods and when utilized together provide complementary clot strength and fiber structural information across diverse clotting conditions.