Fibrin Formation: The Role of the Fibrinogen-Fibrin Monomer Complex

1976 ◽  
Vol 36 (01) ◽  
pp. 037-048 ◽  
Author(s):  
Eric P. Brass ◽  
Walter B. Forman ◽  
Robert V. Edwards ◽  
Olgierd Lindan

SummaryThe process of fibrin formation using highly purified fibrinogen and thrombin was studied using laser fluctuation spectroscopy, a method that rapidly determines particle size in a solution. Two periods in fibrin clot formation were noted: an induction period during which no fibrin polymerization occurred and a period of rapid increase in particle size. Direct measurement of fibrin monomer polymerization and fibrinopeptide release showed no evidence of an induction period. These observations were best explained by a kinetic model for fibrin clot formation incorporating a reversible fibrinogen-fibrin monomer complex. In this model, the complex serves as a buffer system during the earliest phase of fibrin formation. This prevents the accumulation of free polymerizable fibrin monomer until an appreciable amount of fibrinogen has reacted with thrombin, at which point the fibrin monomer level rises rapidly and polymerization proceeds. Clinically, the complex may be a homeostatic mechanism preventing pathological clotting during periods of elevated fibrinogen.

Blood ◽  
1978 ◽  
Vol 52 (4) ◽  
pp. 654-658 ◽  
Author(s):  
EP Brass ◽  
WB Forman ◽  
RV Edwards ◽  
O Lindan

Abstract Using laser fluctuation spectroscopy, a technique that measures particle size change in solution, the kinetics of fibrin clot formation from fibrinogen can be studied. With this technique the effect of calcium on the three distinguishable phases of clot formation, (1) proteolysis of fibrinogen, (2) fibrinogen-fibrin monomer complex formation, and (3) fibrin monomer polymerization, were investigated. Only a small change in the length of the induction period that results from the fibrinogen-fibrin monomer interactions was observed. However, there was a marked increase in the rate of fibrin monomer polymerization in the presence of calcium ions. These data show that calcium decreases the time required for fibrin formation from fibrinogen by markedly accelerating the phase of fibrin monomer polymerization.


Blood ◽  
1978 ◽  
Vol 52 (4) ◽  
pp. 654-658 ◽  
Author(s):  
EP Brass ◽  
WB Forman ◽  
RV Edwards ◽  
O Lindan

Using laser fluctuation spectroscopy, a technique that measures particle size change in solution, the kinetics of fibrin clot formation from fibrinogen can be studied. With this technique the effect of calcium on the three distinguishable phases of clot formation, (1) proteolysis of fibrinogen, (2) fibrinogen-fibrin monomer complex formation, and (3) fibrin monomer polymerization, were investigated. Only a small change in the length of the induction period that results from the fibrinogen-fibrin monomer interactions was observed. However, there was a marked increase in the rate of fibrin monomer polymerization in the presence of calcium ions. These data show that calcium decreases the time required for fibrin formation from fibrinogen by markedly accelerating the phase of fibrin monomer polymerization.


2020 ◽  
Vol 3 (4) ◽  
pp. 67
Author(s):  
Julie Brogaard Larsen ◽  
Anne-Mette Hvas

Disturbance in the balance between fibrin formation and fibrinolysis can lead to either bleeding or thrombosis; however, our current routine coagulation assays are not sensitive to altered fibrinolysis. The clot formation and lysis assay is a dynamic plasma-based analysis that assesses the patient’s capacity for fibrin formation and fibrinolysis by adding an activator of coagulation as well as fibrinolysis to plasma and measuring ex vivo fibrin clot formation and breakdown over time. This assay provides detailed information on the fibrinolytic activity but is currently used for research only, as the assay is prone to inter-laboratory variation and as it demands experienced laboratory technicians as well as specialized personnel to validate and interpret the results. Here, we describe a protocol for the clot formation and lysis assay used at our research laboratory.


1967 ◽  
Vol 126 (5) ◽  
pp. 979-988 ◽  
Author(s):  
B. Lipiński ◽  
J. Hawiger ◽  
J. Jeljaszewicz

Clumping reaction, using standard suspension of Staph. aureus Newman D-2-C strain and various substrates, was quantitatively tested. It has been shown that clumping occurs in fibrin lysate containing soluble fibrin monomer complexes unclottable by thrombin. The reaction was positive with staphylococcal strains possessing clumping factor regardless of staphylocoagulase production. Clumping reaction is similar to paracoagulation reaction induced by protamine sulfate. The substrate for both reactions is stable at 56°C but is destroyed at 60°C. The kinetics of substrate formation for both reactions during fibrin clot lysis is also similar. Clumping reaction with a strain of Staph. epidermidis possessing no clumping factor was positive when these bacteria were coated with protamine sulfate. The effect of heparin, sodium citrate, urea, 2-mercaptoethanol, merthiolate, and mucin on both reactions was tested. The present findings explain the clumping reaction in serum and emphasize the role of blood clotting and fibrinolytic systems in this phenomenon.


2014 ◽  
Vol 111 (05) ◽  
pp. 842-850 ◽  
Author(s):  
Cédric Duval ◽  
Peter Allan ◽  
Simon D. A. Connell ◽  
Victoria C. Ridger ◽  
Helen Philippou ◽  
...  

SummaryFactor XIII is responsible for the cross-linking of fibrin γ-chains in the early stages of clot formation, whilst α-chain cross-linking occurs at a slower rate. Although γ- and α-chain cross-linking was previously shown to contribute to clot stiffness, the role of cross-linking of both chains in determining clot structure is currently unknown. Therefore, the aim of this study was to determine the role of individual α- and γ-chain cross-linking during clot formation, and its effects on clot structure. We made use of a recombinant fibrinogen (γQ398N/Q399N/K406R), which does not allow for y-chain cross-linking. In the absence of cross-linking, intact D-D interface was shown to play a potential role in fibre appearance time, clot stiffness and elasticity. Cross-linking of the fibrin α-chain played a role in the thickening of the fibrin fibres over time, and decreased lysis rate in the absence of α2-antiplasmin. We also showed that α-chain cross-linking played a role in the timing of fibre appearance, straightening fibres, increasing clot stiffness and reducing clot deformation. Cross-linking of the γ-chain played a role in fibrin fibre appearance time and fibre density. Our results show that α- and γ-chain cross-linking play independent and specific roles in fibrin clot formation and structure.


2021 ◽  
Vol 27 ◽  
pp. 107602962110185
Author(s):  
Reheman Adili ◽  
Madeline Jackson ◽  
Livia Stanger ◽  
Xiangrong Dai ◽  
Mandy Li ◽  
...  

Uncontrolled bleeding associated with trauma and surgery is the leading cause of preventable death. Batroxobin, a snake venom-derived thrombin-like serine protease, has been shown to clot fibrinogen by cleaving fibrinopeptide A in a manner distinctly different from thrombin, even in the presence of heparin. The biochemical properties of batroxobin and its effect on coagulation have been well characterized in vitro. However, the efficacy of batroxobin on hemostatic clot formation in vivo is not well studied due to the lack of reliable in vivo hemostasis models. Here, we studied the efficacy of batroxobin and slounase, a batroxobin containing activated factor X, on hemostatic clot composition and bleeding using intravital microcopy laser ablation hemostasis models in micro and macro vessels and liver puncture hemostasis models in normal and heparin-induced hypocoagulant mice. We found that prophylactic treatment in wild-type mice with batroxobin, slounase and activated factor X significantly enhanced platelet-rich fibrin clot formation following vascular injury. In heparin-treated mice, batroxobin treatment resulted in detectable fibrin formation and a modest increase in hemostatic clot size, while activated factor X had no effect. In contrast, slounase treatment significantly enhanced both platelet recruitment and fibrin formation, forming a stable clot and shortening bleeding time and blood loss in wild-type and heparin-treated hypocoagulant mice. Our data demonstrate that, while batroxobin enhances fibrin formation, slounase was able to enhance hemostasis in normal mice and restore hemostasis in hypocoagulant conditions via the enhancement of fibrin formation and platelet activation, indicating that slounase is more effective in controlling hemorrhage.


Author(s):  
Б.И. Кузник ◽  
С.О. Давыдов ◽  
Е.С. Гусева ◽  
Ю.Н. Смоляков ◽  
А.В. Степанов ◽  
...  

Цель - изучение роли отдельных форменных элементов крови в развитии гемокоагуляционного потенциала у здоровых женщин и, больных гипертонической болезнью (ГБ). Методика. В исследовании приняли участие 102 женщины. Контрольную группу составили 30 относительно здоровых женщин. Больные ГБ были разделены на 2 подгруппы: в одну (ГБ-1) вошли 37 женщин с гипертонической болезнью II стадии, в другую (ГБ-2) - 35 женщин с ГБ II стадии, регулярно проходящих на протяжении 2-3 лет по 3-4 курса кинезитерапии. Определяли число тромбоцитов, активированное частичное тромбопластиновое время (АЧТВ), протромбиновое время (ПВ), тромбиновое время (ТВ), концентрацию фибриногена и пространственный рост фибринового сгустка, включающий время задержки роста сгустка, начальную и стационарную скорость его роста, плотность и размер сгустка. С помощью корреляционного анализа оценивалась роль отдельных форменных элементов крови в развитии гемокоагуляционного потенциала у здоровых и больных ГБ женщин. Результаты. У всех обследованных женщин обнаружено увеличение числа лимфоцитов и эозинофилов, возрастание предрасположенности к тромбообразованию, выявляемое с помощью оценки тромбодинамических свойств сгустка, возрастание скорости формирования и размеры фибринового сгустка. У женщин в группе ГБ-2 эти сдвиги выражены в меньшей степени. У здоровых женщин обнаружены прямые корреляции между числом моноцитов, АЧТВ и тромбиновым временем. и отрицательные - между числом лимфоцитов и АЧТВ. Число эозинофилов у здоровых женщин положительно коррелирует с протромбиновым временем и плотностью сгустка. В ГБ-1 отмечается негативная связь между числом нейтрофилов и скоростью образования сгустка, а также положительная связь между числом моноцитов, скоростью и размером сгустка и между числом базофилов и тромбиновым временем. У больных ГБ-2, принимавших кинезитерапию число эритроцитов отрицательно коррелирует с АЧТВ, количество тромбоцитов обнаруживает положительную корреляцию тромбиновым временем, скоростью и размерами сгустка, а общее число лейкоцитов - с протромбиновым временем и скоростью образования сгустка. Содержание нейтрофилов положительно коррелирует с протромбиновым временем и отрицательно со скоростью образования сгустка. Количество лимфоцитов и эозинофилов отрицательно коррелирует со скоростью образования сгустка, а базофилов - с уровнем фибриногена и скоростью появления сгустка. Заключение. В формировании гемокоагуляционного потенциала у здоровых женщин и больных ГБ ведущая роль принадлежит тромбоцитам и различным популяциям лейкоцитов. Обсуждается положительное влияние кинезитерапии. Aim. To study the role of different blood cells in the development of coagulation potential in healthy women and patients with essential hypertension (EH). Methods. The study included 102 women. The control group consisted of 30 relatively healthy women. Patients with EH were divided into 2 subgroups: the first subgroup (EH-1) included 37 women with stage II arterial hypertension, the second subgroup (EH-2) - 35 women with stage II EH who received 3-4 courses of kinesitherapy for 2-3 years on a regular basis. The following values were determined: platelet count, activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), fibrinogen concentration, and spatial fibrin clot growth, including the delay time of clot lengthening, initial and steady growth rate, and clot density and size. The role of different blood cells in the development of coagulation potential was evaluated in healthy and EH women using the correlation analysis. Results. All women had increased numbers of lymphocytes and eosinophils; showed a predisposition to thrombosis as was evident from thrombodynamic properties of the clot; and increased formation rate and size of the fibrin clot. These changes were less pronounced in the EH-2 group. In healthy women, a direct correlation was observed between the number of monocytes, APTT and the thrombin time. and a negative correlation - between the number of lymphocytes and APTT. In this group, the number of eosinophils positively correlated with the prothrombin time and the clot density. In the EH-1 group, the number of neutrophils inversely correlated with the rate of clot formation; the number of monocytes positively correlated with the clot formation rate and size; and the number of basophils positively correlated with the thrombin time. In EH-2 patients receiving kinesitherapy, the number of red cells inversely correlated with APTT; the number of platelets positively correlated with the thrombin time, the clot formation rate and size; and the total number of leukocytes positively correlated with the prothrombin time and the clot formation rate. The neutrophil count positively correlated with the prothrombin time and negatively - with the rate of clot formation. The number of lymphocytes and eosinophils negatively correlated with the rate of clot formation, and the number of basophils - with the fibrinogen level and the rate of clot emergence. Conclusion. Platelets and leukocyte populations play the main role in the formation of coagulation potential in healthy women and patients with EH. The beneficial effect of kinesitherapy is discussed.


1972 ◽  
Vol 28 (02) ◽  
pp. 244-256 ◽  
Author(s):  
T. Z Muzaffar ◽  
G. G Youngson ◽  
W. A. J Bryce ◽  
D. P Dhall

SummaryIn this study highly reproducible turbidimetric techniques are used to investigate clotting of human platelet-poor plasma and purified human fibrinogen solutions by thrombin. Three phases are clearly distinguishable turbidimetrically during clotting. All the three phases are altered by dextran. This effect of dextran is not mediated by any action on the enzymatic activity of thrombin. Further, it is independent of the molecular weight of dextran, but is found related to the final dextran, thrombin and fibrinogen concentrations. Evidence is presented to show that dextran accelerates the polymerisation of fibrin monomer. However, it seems clear that dextran has an additional action quite apart from this effect. These actions of dextran are discussed in relation to kinetics of clot formation.


1981 ◽  
Author(s):  
T J Frost ◽  
R S Lau

The cascade mechanism for the coagulation pathway has prevailed since its original proposal, and subsequent observations have supported the role of thrombin on coagulation reactions other than in Fibrinogen - Fibrin conversion. In very small quantities, thrombin has been reported as increasing the activity of Factor VIII: C. We report here, observations which support a reverse proposal, namely that Factor VIII: C increases activity of thrombin in Fibrinogen - Fibrin conversion.Thrombin clotting times were performed employing human Fibrinogen solutions, fresh citrated normal platelet - poor human plasma and fresh citrated platelet - poor plasma from severely affected Hemophilia A patients, in whom Factor VIII : C assays were less than 1% in a one stage, activated partial thromboplastin time assay. A platelet aggregometer was used to assess the thrombin time, and the rate of clot formation (fibrin polymerization) was indicated by optical density change in the form of a curve, the tangent of the maximal slope of curve being equated with this clot formation rate.Results indicate that with very small concentrations of thrombin, Factor VIII: C enhances the rate of fibrin clot formation and that Factor VIII: C restores the abnormal rate of clot formation observed in severe Hemophilia A; the test system was known to be calcium-free, which ensured that activation of Factor X did not ensue. The concentration of Fibrinogen remained constant in all experiments. The observed phenomena are confirmed when employing purified fibrinogen solutions, using decreasing amounts of Factor VIII: C.


1977 ◽  
Author(s):  
Fletcher B. Taylor

The dilute whole blood clot lysis assay has been used in diagnosis of patients with deep vein thrombophlebitis and pulmonary embolism. Because of its clinical value it has also been the subject of biochemical and physiologic studies of clot lysis of normal diluted blood. This assay reflects the behavior of platelets in that clot lysis as well as clot retraction are platelet dependent. Further, this contribution of platelets is temperature sensitive whereas the rate of fibrin clot formation is not. Thus, this assay offers a convenient model for functional and morphologic studies of temperature induced discontinuity of platelet-fibrin assembly and the interaction of platelet and fibrin in clot formation, retraction and lysis. In these studies the release of serotonin from platelets was correlated with clot formation, retraction, lysis, and clot morphology at 1) 37°, 4° and 4°−37° at 5, 15, 30 and 60 minute intervals. The results suggest that both platelets and fibrinogen are influenced by thrombin used in the assay and that under certain temperature conditions the platelets will release their contents out of phase with the assembly of the fibrin network. In cases where this discontinuity exists, the diluted clots will not retract or lyse normally.


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