Carbon monoxide releasing molecule-2 increases the velocity of thrombus growth and strength in hemophilia A, hemophilia B and factor VII-deficient plasmas

Abstract

Carbon monoxide derived from carbon monoxide releasing molecules (CORMs) has been demonstrated to enhance normal plasma thrombus speed of growth and strength in vitro. We tested the hypothesis that tricarbonyldichlororuthenium (II) dimer (CORM-2) improves the velocity of formation and strength of hemophiliac plasma thrombi as determined by thrombelastography. Plasma deficient (<1% normal activity) in factor VIII (FVIII; n = 11 individuals), factor IX (FIX; n = 5 individuals) or factor VII (FVII; n = 4 individuals) was exposed to 0 or 100 micromol CORM-2, with coagulation initiated with tissue factor. Coagulation kinetics were monitored with thrombelastography for 15 min. Paired t-tests were used to analyze FVIII-deficient plasma results; relative change was used to describe the other plasma types tested. In FVIII-deficient plasma, CORM-2 exposure significantly (P < 0.05) increased the velocity of thrombus formation (84%) and strength (48%) compared with plasma not exposed to CORM-2. FXI-deficient clots demonstrated an increase in velocity of formation (63%) and strength (43%) after CORM-2 exposure. Lastly, CORM-2 exposure increased FVII-deficient plasma velocity of formation (45%) and strength (63%). CORM-2 markedly enhanced the velocity of clot growth and strength in hemophiliac plasma. These findings serve as the rationale to determine whether CORMs could be utilized as hemostatic agents.

Carbon monoxide (CO)-releasing molecule-derived CO regulates tissue factor and plasminogen activator inhibitor type 1 in human endothelial cells

“…CORM-2 increases the velocity of thrombus growth and strength in hemophilia A, hemophilia B, and factor VII-deficient plasmas [36]. Furthermore, CORM-2 enhances coagulation and markedly attenuates fibrinolysis in human plasma [37].…”

Carbon monoxide: Anticoagulant or procoagulant?

“…were exposed to CORM-2 [51][52][53][54][55][56][57] In order to further characterize the mechanism by which CO enhanced coagulation, experiments involving isolated exposure of prothrombin [58] or fibrinogen [59] to CORM-2 were performed, with the purified protein function assessed in prothrombin or fibrinogen deficient plasmas, respectively. These works lead to the determination that only fibrinogen was affected by CO [58,59], and additional work utilizing mass spectrometry [60] demonstrated that protease digestion of fibrinogen was modified by exposure to CO. More importantly, it was noted that a heme group(s) was present in the fibrinogen digest samples, and following exposure of human plasma to compounds that either produced a carboxyheme or metheme state, plasma strength was increased or decreased respectively, indicative of heme-based modulation of fibrinogen function [60].…”

“…4), with methods used to generate these images as previously cited [84][85][86]. Considered together, these investigations involved conditions or disease states wherein increased CO would be expected to be present, resulting in thrombus ultrastructure (e.g., finer or matted fibers) consistent with enhanced clot strength demonstrated in vitro [48,[51][52][53][54][55][56][57]67,68,[70][71][72][73][74][75][76] or in vivo [68,69] following CO exposure.…”

“…Platelets appear to respond to CO in a similar fashion among human and rodent studies [10][11][12]16,21,22], but plasmatic coagulation also appears to be enhanced in humans, rats and rabbits [48,[51][52][53][54][55][56][57][58][67][68][69]. In an effort to reveal potential sources of specie specific differences, one may consider the differences in thrombus ultrastructure between humans, rabbits, mice and rats [94,95].…”

Carbon Monoxide Releasing Molecule-2 Improves Protamine-Mediated Hypocoagulation/Hyperfibrinolysis in Human Plasma In Vitro

“…Concordantly, electron microscopy revealed that thin, strong fibrin fibers (which are also more resistant to fibrinolysis) form to a greater extent in plasma thrombi than thick, weaker fibers in the presence of CORM-2 [14]. Studies with hemophiliac plasma [15], plasma obtained from subjects administered warfarin [16], and plasma diluted with crystalloid or colloid [17] demonstrated improved coagulation kinetics and decreased vulnerability to lysis following exposure to CORM-2. In sum, rather than increase thrombin generation, CORM-2 modifies the substrate characteristics of fibrinogen, thus enhancing coagulation and attenuating fibrinolysis.…”