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Biomechanics

Biomechanics - Poster Session A

Poster D10 - Assessing the impact of blood plasma components on clot mechanical properties

Thursday, October 24, 2024
10:00 AM – 11:00 AM EST
Location: Exhibit Hall E, F & G

Introduction:  The dysregulation of the coagulation cascade following traumatic injury can result in excessive bleeding. Trauma can result in changes in blood composition. Increased release of tissue factor, a key protein that initiates blood clotting, occurs after trauma ruptures tissue. Fibrinogen, the main structural protein in blood clots, is depleted due to excessive bleeding, diluted by treatment with IV fluids, and the consumption of clotting factors. Following the initiation of clotting, fibrinogen is transformed into fibrin, which creates the network structure essential for giving blood clots their stability to stop bleeding. The structural integrity of blood clots can be measured by their burst pressure, which reflects the maximum force the clot can withstand before rupturing. We aim to investigate how changes in blood composition affect the structural stability of blood plasma clots.

Materials and

Methods:  Fibrinogen and tissue factor concentrations were varied to assess their influence on blood clot mechanics. Clots were formed with human pooled plasma, 20 mM calcium chloride solution, and tissue factor. The fibrinogen concentration in plasma was adjusted by diluting the plasma with saline to achieve 15% and 30% dilutions (2.0 mg/mL and 1.7 mg/mL fibrinogen respectively). Plasma clots were formed within 3D-printed discs, creating a clot with a 20 mm diameter and 4 mm thickness. The discs with clots were fastened to a biomomentum mechanical tester that applied force perpendicular to the surface of the plasma clot using a 2 mm pin while burst force to break each clot was measured and recorded. Confocal microscopy was used to image the fibrin networks of clots, and ImageJ software was utilized to calculate the fiber density of these networks. This method allowed us to visualize and quantify the structural characteristics of the fibrin clots. By comparing fibrin fiber density with the collected burst pressure data, the relationship between the microstructural properties of the fibrin network and the mechanical stability of the clots can be determined.

Results, Conclusions, and Discussions: Burst pressure varied among the clots tested, with higher concentrations of tissue factor resulting in increased burst pressures (Figure A) and lower concentrations of fibrinogen leading to decreased burst pressures (Figure B). Significant differences in clot burst forces were observed between the highest tissue factor concentration (average burst pressure of 31.84±15.75 gf) and the lowest fibrinogen concentration (average burst pressure of 14.44±4.45 gf) (Figure C). A significant correlation was noted between increasing fibrin fiber density and burst force. Clots with the highest tissue factor concentration had an average fiber density of 28.37±1.35%, while those with the lowest fibrinogen concentration had an average fiber density of 19.67±2.75% (Figure D). There was a direct relationship between burst force and fiber density (p < 0.05).

The concentration of tissue factor and fibrinogen is directly related to clot fibrin fiber density and the ability to withstand rupture. Higher levels of tissue factor will accelerate and amplify the coagulation process, leading to more thrombin production and faster fibrin polymerization, thereby creating a denser fibrin network within the clot. When fibrinogen levels are low, there is a reduction in fibrinogen molecules that can be converted into fibrin for the clot fibrin network. Simulating trauma-induced changes shows that alterations in clot components weaken structural stability, causing clot failure.

Key steps for research on trauma and blood clot stability include conducting tests with whole blood samples to better simulate physiological conditions. Additionally, integrating the testing of therapeutic interventions, such as fibrinogen concentrates, cryoprecipitate, and fresh frozen plasma, will help enhance fibrinogen levels and stabilize clots after formation.

Acknowledgements (Optional):  This research was supported by the National Science Foundation: EEC-1950509 and EEC-2349225.