Ukr.Biochem.J. 2021; Volume 93, Issue 2, Mar-Apr, pp. 62-70

doi: https://doi.org/10.15407/ubj93.02.062

Thromboelastographic study of fibrin clot and molecular basis of maximum clot firmness

18.05.2021   Uncategorized   No comments

D. S. Korolova1*, Y. M. Stohnii1, V. I. Gryshchuk1, S. I. Zhuk2,
I. V. Us2, T. M. Chernyshenko1, O. P. Kostiuchenko1, K. P. Klymenko1,
O. M. Platonov1,3, O. I. Ivashchenko3, V. O. Chernyshenko1

1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Shupyk National Medical Academy of Postgraduate Education, Kyiv, Ukraine;
3ESC “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Ukraine;
e-mail: d.korolova@gmail.com

Received: 29 January 2021; Accepted: 23 April 2021

Maximum clot firmness (MCF) is the main parameter of thromboelastography (TEG) reflecting the stability of a clot. In this work, we looked for markers that can influence the enhancement of MCF and detec­ted molecular markers and blood clotting parameters that can be involved in such mechanisms. Blood samples of pregnant women with placental disorders were collected in the Kyiv Perinatal Center. TEG was performed on whole blood in EXTEM and INTEM tests. APTT, INR, fibrinogen concentration and platelet aggregation were measured using traditional laboratory approaches. D-dimer was detected in sandwich ELISA using monoclonal antibodies III-3B and II-4D. The relative cross-linking activity of factor XIIIa was measured by the direct quantification of the cross-linked γ-chain of fibrin using Western-Blotting with monoclonal antibody II-4D. D-dimer and fibrinogen concentrations, clotting time in the APTT test, INR and rate of platelet aggregation did not correlate with the MCF. However, we found positive correlations of MCF with factor XIIIa activity: 0.51 and 0.87 for EXTEM and INTEM, respectively. These data indicate that for normal and slightly increased fibrinogen concentrations, fibrin clot firmness will depend mostly on the activity of factor XIIIa. Thus the direct determination of factor XIIIa activity in blood plasma of patients can be relevant for predicting the risk of intravascular coagulation. Evaluation of the content and activity of individual clotting factors or other components of the coagulation system can be useful additions to the TEG diagnostics and should not be neglected.

Keywords: , , , ,

References:

  1. Othman M, Kaur H. Thromboelastography (TEG). Methods Mol Biol. 2017;1646:533-543.  PubMed, CrossRef
  2. Somani V, Amarapurkar D, Shah A. Thromboelastography for assessing the risk of bleeding in patients with cirrhosis-moving closer. J Clin Exp Hepatol. 2017;7(4):284-289. PubMed, PubMedCentral, CrossRef
  3. He Y, Xin X, Geng Y, Tang N, Zhou J, Li D. The value of thromboelastography for bleeding risk prediction in hematologic diseases. Am J Med Sci. 2016;352(5):502-506. PubMed, CrossRef
  4. Akay OM. The double hazard of bleeding and thrombosis in hemostasis from a clinical point of view: a global assessment by rotational thromboelastometry (ROTEM). Clin Appl Thromb Hemost. 2018;24(6):850-858. PubMed, PubMedCentral, CrossRef
  5. Kim SM, Kim SI, Yu G, Kim JS, Hong SI, Chae B, Shin YS, Kim YJ, Jang S, Kim WY. Role of Thromboelastography as an Early Predictor of Disseminated Intravascular Coagulation in Patients with Septic Shock. J Clin Med. 2020;9(12):3883. PubMed, PubMedCentral, CrossRef
  6. Wu S, Yuan H, Zhou Y, Long Z, Peng Y, Peng F. Thromboelastography-based assessment of coagulation function in patients with chronic kidney disease and the risk factors of hypercoagulability. Nan Fang Yi Ke Da Xue Xue Bao. 2020;40(4):556-561. (In Chinese). PubMed, PubMedCentral, CrossRef
  7. Raval JS, Burnett AE, Rollins-Raval MA, Griggs JR, Rosenbaum L, Nielsen ND, Harkins MS. Viscoelastic testing in COVID-19: a possible screening tool for severe disease? Transfusion. 2020;60(6):1131-1132. PubMed, PubMedCentral, CrossRef
  8. Pavoni V, Gianesello L, Pazzi M, Stera C, Meconi T, Frigieri FC. Evaluation of coagulation function by rotation thromboelastometry in critically ill patients with severe COVID-19 pneumonia. J Thromb Thrombolysis. 2020;50(2):281-286. PubMed, PubMedCentral, CrossRef
  9. Liu C, Guan Z, Xu Q, Zhao L, Song Y, Wang H. Relation of thromboelastography parameters to conventional coagulation tests used to evaluate the hypercoagulable state of aged fracture patients. Medicine (Baltimore). 2016;95(24):e3934. PubMed, PubMedCentral, CrossRef
  10. Chandrashekar A, Singh G, Garry J, Sikalas N, Labropoulos N. Mechanical and Biochemical Role of Fibrin Within a Venous Thrombus. Eur J Vasc Endovasc Surg. 2018;55(3):417-424. PubMed, CrossRef
  11. Xu S, Xu Z, Kim OV, Litvinov RI, Weisel JW, Alber M. Model predictions of deformation, embolization and permeability of partially obstructive blood clots under variable shear flow. J R Soc Interface. 2017;14(136):20170441. PubMed, PubMedCentral, CrossRef
  12. Lugovskoi EV, Kolesnikova IN, Komisarenko SV. Usage of monoclonal antibodies for determination of localization of antigenic determinants and fibrin polymerization sites within fibrinogen and fibrin molecules and their application in test–systems for diagnostics and the threat of thrombus formation. Biotechnologia Acta. 2013; 6(4): 33-42. CrossRef
  13. Platonova TN, Sushko OO, Colovjov DO, Iena IaM. Determination of fibrinogen content in human blood plasma using thrombin-like enzyme of ancistron N and analysis of hemostasis under presence of blood coagulation inhibitors. Fiziol Zh. 1993;39(1):15-19. (In Ukrainian). PubMed
  14. Sokolovska AS, Chernyshenko TM, Ivanenko TI. Comparative characteristic of fibrinogen level determination methods in blood plasma. Exp Clin Physiol Biochem. 2002; 3: 82-86. (In Ukrainian).
  15. Lugovskoi EV, Kolesnikova IN, Lugovskaia NE, Litvinova LM, Gritsenko PG, Gogolinskaia GK, Liashko ED, Kostiuchenko EP, Remizovskiy GA, Pedchenko VN, Komisarenko SV. Quantification of D-dimer and soluble fibrin in blood plasma of people with ischemic heart disease and hypertension. Ukr Biokhim Zhurn. 1999; 76(6): 136-141. (In Russian). PubMed
  16. Born GV. Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature. 1962;194:927-929. PubMed, CrossRef
  17. He F. Laemmli-SDS-PAGE. Bio-101. 2011:e80.  CrossRef
  18. Wu HC, Yen CC, Tsui WH, Chen HM. A red line not to cross: evaluating the limitation and properness of gel image tuning procedures. Anal Biochem. 2010;396(1):42-50. PubMed, CrossRef
  19. Howie PW. Blood clotting and fibrinolysis in pregnancy. Postgrad Med J. 1979;55(643):362-366. PubMed, PubMedCentral, CrossRef
  20. Pat. 138359UA, ICP G 01 N33/00. The method of quantitative determination of enzymatic activity of bacterial Ca2+-independent transglutaminase  / Gryshchuk V.I., Stohnii  Y.M., Chernyshenko V.O., Chernyshenko T.M., Lugovska N.E. P. Publ. 14.05.2019, Bul. N 22. (In Ukrainian).
  21. Katona É, Pénzes K, Molnár É, Muszbek L. Measurement of factor XIII activity in plasma. Clin Chem Lab Med. 2012;50(7):1191-1202. PubMed, CrossRef
  22. Hellstern P, Schilz K, von Blohn G, Wenzel E. Determination of factor XIII activity and of factor XIII inhibitors using an ammonium-sensitive electrode. Thromb Haemost. 1983;50(2):563-566. PubMed, CrossRef
  23. Pabinger I, Grafenhofer H. Thrombosis during pregnancy: risk factors, diagnosis and treatment. Pathophysiol Haemost Thromb. 2002;32(5-6):322-324. PubMed, CrossRef
  24. Lykke JA, Bare LA, Olsen J, Lagier R, Arellano AR, Tong C, Paidas MJ, Langhoff-Roos J. Thrombophilias and adverse pregnancy outcomes: results from the Danish National Birth Cohort. J Thromb Haemost. 2012;10(7):1320-1325. PubMed, CrossRef
  25. Rodger MA. An update on thrombophilia and placenta mediated pregnancy complications: what should we tell our patients? Thromb Res. 2013;131(Suppl 1):S25-S27. PubMed, CrossRef
  26. Weisel JW, Litvinov RI. Fibrin Formation, Structure and Properties. Subcell Biochem. 2017;82:405-456. PubMed, PubMedCentral, CrossRef
  27. Lam WA, Chaudhuri O, Crow A, Webster KD, Li TD, Kita A, Huang J, Fletcher DA. Mechanics and contraction dynamics of single platelets and implications for clot stiffening. Nat Mater. 2011;10(1):61-66. PubMed, PubMedCentral, CrossRef
  28. Palta S, Saroa R, Palta A. Overview of the coagulation system. Indian J Anaesth. 2014;58(5):515-523. PubMed, PubMedCentral, CrossRef
  29. Lippi G, Cervellin G, Franchini M, Favaloro EJ. Biochemical markers for the diagnosis of venous thromboembolism: the past, present and future. J Thromb Thrombolysis. 2010;30(4):459-471. PubMed, CrossRef
  30. Peng HT, Nascimento B, Beckett A. Thromboelastography and thromboelastometry in assessment of fibrinogen deficiency and prediction for transfusion requirement: A descriptive review. Biomed Res Int. 2018;2018:7020539. PubMed, PubMedCentral, CrossRef
Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License.