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Thromb Haemost 2014; 111(04): 656-661
DOI: 10.1160/TH13-06-0479
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Improvement of fibrin clot structure after factor VIII injection in haemophilia A patients treated on demand

Aleksandra Antovic
1   Karolinska Institutet, Department of Clinical Sciences, Danderyd Hospital, Division of Cardiovascular Medicine, Stockholm, Sweden
,
Danijela Mikovic
2   Haemostasis Department and Haemophilia Centre, Blood Transfusion Institute of Serbia, Belgrade, Serbia
,
Ivo Elezovic
3   Clinic of Haematology, Clinical Centre of Serbia & Faculty of Medicine, University of Belgrade, Serbia
,
Michael Zabczyk
4   Department of Molecular Medicine and Surgery/Clinical Chemistry/Coagulation Research, Karolinska Institute, Stockholm, Sweden
5   Institute of Cardiology, Medical College, Jagiellonian University, Krakow, Poland
,
Kjell Hutenby
6   Division of Clinical Research, Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden
,
Jovan P. Antovic
7   Department of Coagulation Research, Institute for Molecular Medicine and Surgery, Karolinska Institutet, Stockholm, Sweden
8   Department of Clinical Chemistry, Karolinska University Hospital, Stockholm, Sweden
› Author Affiliations
Further Information

Publication History

Received: 13 June 2013

Accepted after major revision: 23 October 2013

Publication Date:
29 November 2017 (online)

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Summary

Patients with haemophilia A have seriously impaired thrombin generation due to an inherited deficiency of factor (F)VIII, making them form unstable fibrin clots that are unable to maintain haemostasis. Data on fibrin structure in haemophilia patients remain limited. Fibrin permeability, assessed by a flow measurement technique, was investigated in plasma from 20 patients with severe haemophilia A treated on demand, before and 30 minutes after FVIII injection. The results were correlated with concentrations of fibrinogen, FVIII and thrombin-activatable fibrinolysis inhibitor (TAFI), and global haemostatic markers: endogenous thrombin potential (ETP) and overall haemostatic potential (OHP). Fibrin structure was visualised using scanning electron microscopy (SEM). The permeability coefficient Ks decreased significantly after FVIII treatment. Ks correlated significantly with FVIII levels and dosage, and with ETP, OHP and levels of TAFI. SEM images revealed irregular, porous fibrin clots composed of thick and short fibers before FVIII treatment. The clots had recovered after FVIII replacement almost to levels in control samples, revealing compact fibrin with smaller intrinsic pores. To the best of our knowledge, this is the first description of fibrin porosity and structure before and after FVIII treatment of selected haemophilia patients. It seems that thrombin generation is the main determinant of fibrin structure in haemophilic plasma.

Keywords

Fibrin clot structure - scanning electron microscopy - FVIII - haemophilia A
 

  • References

  • 1 Dargaud Y, Beguin S, Lienhart A. et al. Evaluation of thrombin generating capacity in plasma from patients with haemophilia A and B.. Thromb Haemost 2005; 93: 475-480.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 2 Ramström G, Blombäck M, Egberg N. et al. Oral surgery in patients with hereditary bleeding disorders. A survey of treatment in the Stockholm area (1974-1985).. Int J Oral Maxillofac Surg 1989; 18: 320-322.
    CrossrefPubMedGoogle Scholar
  • 3 Blanchette VS, Manco-Johnson M, Santagostino E. et al. Optimizing factor prophylaxis for the haemophilia population: where do we stand?. Haemophilia 2004; 10 (Suppl. 04) 97-104.
    CrossrefPubMedGoogle Scholar
  • 4 Blomback B, Carlsson K, Fatah K. et al. Fibrin in human plasma: gel architectures governed by rate and nature of fibrinogen activation.. Thromb Res 1994; 75: 521-538.
    CrossrefPubMedGoogle Scholar
  • 5 Rainsford SG, Hall A. A three-year study of adolescent boys suffering from haemophilia and allied disorders.. Br J Haematol 1973; 24: 539-551.
    CrossrefPubMedGoogle Scholar
  • 6 Aledort LM, Haschmeyer RH, Pettersson H. A longitudinal study of orthopaedic outcomes for severe factor-VIII-deficient haemophiliacs. The Orthopaedic Outcome Study Group.. J Intern Med 1994; 236: 391-399.
    CrossrefPubMedGoogle Scholar
  • 7 Wolberg AS, Allen GA, Monroe DM. et al. High dose factor Vila improves clot structure and stability in a model of haemophilia B.. Br J Haematol 2005; 131: 645-655.
    CrossrefPubMedGoogle Scholar
  • 8 He S, Blombäck M, Jacobsson Ekman G. et al. The role of recombinant factor Vila (FVIIa) in fibrin structure in the absence of FVIII/FIX.. J Thromb Haemost 2003; 1: 1215-1219.
    CrossrefPubMedGoogle Scholar
  • 9 Dargaud Y, Prevost C, Lienhart A. et al. Evaluation of the overall haemostatic effect of recombinant factor VIIa by measuring thrombin generation and stability of fibrin clots.. Haemophilia 2011; 17: 957-961.
    CrossrefPubMedGoogle Scholar
  • 10 Brummel-Ziedins KE, Branda RF, Butenas S, Mann KG. Discordant fibrin formation in haemophilia.. J Thromb Haemost 2009; 7: 825-832.
    CrossrefPubMedGoogle Scholar
  • 11 Antovic JP, Mikovic D, Elezovic I. et al. Two global haemostatic assays as additional tools to monitor treatment in cases of Haemophilia A.. Thromb Haemost 2012; 108: 21-31.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 12 Blomback B, Carlsson K, Hessel B. et al. Native fibrin gel networks observed by 3D microscopy, permeation and turbidity.. Biochim Biophys Acta 1989; 997: 96-110.
    CrossrefPubMedGoogle Scholar
  • 13 He S, Cao H, Antovic A. et al. Modifications of flow measurement to determine fibrin gel permeability and the preliminary use in research and clinical materials.. Blood Coagul Fibrinolysis 2005; 16: 61-67.
    CrossrefPubMedGoogle Scholar
  • 14 Berntorp E, Björkman S.. The pharmacokinetics of clotting factor therapy.. Haemophilia 2003; 9: 353-359.
    CrossrefPubMedGoogle Scholar
  • 15 Carr Jr. ME, Shen LL, Hermans J. Mass-length ratio of fibrin fibres from gel permeation and light scattering.. Biopolymers 1977; 16: 1-15.
    CrossrefPubMedGoogle Scholar
  • 16 Dey S. A new rapid air-drying technique for scanning electron microscopy using tetramethylsilane: application to mammalian tissue.. Cytobios 1993; 73: 17-23.
    PubMedGoogle Scholar
  • 17 Weibel E. Stereological methods: Practical methods for biological morphometry. vol 1. Academic Press; 1979. London: 91-100.
    Google Scholar
  • 18 Mikovic D, Woodhams BJ, Holmström M. et al. On-demand but not prophylactic treatment with FVIII concentrate increases thrombin activatable fibrinolysis inhibitor activation in severe haemophilia A patients.. Int J Lab Hematol 2012; 34: 35-40.
    CrossrefPubMedGoogle Scholar
  • 19 Antovic A. The Overall Haemostasis Potential (OHP): a laboratory tool for the investigation of global haemostasis.. Semin Thromb Haemost 2010; 36: 772-779.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 20 Hyas AM, Sorensen HT, Norengaard L. et al. Tranexamic acid combined with recombinant factor VIII increases clot resistance to accelerated fibrinolysis in severe haemophilia A.. J Thromb Haemost 2007; 5: 2408-2414.
    CrossrefPubMedGoogle Scholar
  • 21 Hoffman M. A cell-based model of coagulation and the role of factor VIIa.. Blood Rev 2003; 17 (Suppl. 01) S1-5.
    CrossrefPubMedGoogle Scholar
  • 22 Blomback B, Hessel B, Hogg D. et al. A two-step fibrinogen--fibrin transition in blood coagulation.. Nature 1978; 275: 501-505.
    CrossrefPubMedGoogle Scholar
  • 23 Hantgan RR, Hermans J. Assembly of fibrin. A light scattering study.. J Biol Chem 1979; 254: 11272-11281.
    CrossrefPubMedGoogle Scholar