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

Aleksandra Antovic; Danijela Mikovic; Ivo Elezovic; Michael Zabczyk; Kjell Hutenby; Jovan P. Antovic

doi:10.1160/TH13-06-0479

Thrombosis and Haemostasis, Inhaltsverzeichnis

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

¹Karolinska Institutet, Department of Clinical Sciences, Danderyd Hospital, Division of Cardiovascular Medicine, Stockholm, Sweden

,

Danijela Mikovic

²Haemostasis Department and Haemophilia Centre, Blood Transfusion Institute of Serbia, Belgrade, Serbia

,

Ivo Elezovic

³Clinic of Haematology, Clinical Centre of Serbia & Faculty of Medicine, University of Belgrade, Serbia

,

Michael Zabczyk

⁴Department of Molecular Medicine and Surgery/Clinical Chemistry/Coagulation Research, Karolinska Institute, Stockholm, Sweden

⁵Institute of Cardiology, Medical College, Jagiellonian University, Krakow, Poland

,

Kjell Hutenby

⁶Division of Clinical Research, Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden

,

Jovan P. Antovic

⁷Department of Coagulation Research, Institute for Molecular Medicine and Surgery, Karolinska Institutet, Stockholm, Sweden

⁸Department of Clinical Chemistry, Karolinska University Hospital, Stockholm, Sweden

› Institutsangaben

Abstract

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

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Referenzen

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.
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.
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.
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.
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.
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.
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.
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.
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.
10 Brummel-Ziedins KE, Branda RF, Butenas S, Mann KG. Discordant fibrin formation in haemophilia.. J Thromb Haemost 2009; 7: 825-832.
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.
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.
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.
14 Berntorp E, Björkman S.. The pharmacokinetics of clotting factor therapy.. Haemophilia 2003; 9: 353-359.
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.
16 Dey S. A new rapid air-drying technique for scanning electron microscopy using tetramethylsilane: application to mammalian tissue.. Cytobios 1993; 73: 17-23.
17 Weibel E. Stereological methods: Practical methods for biological morphometry. vol 1. Academic Press; 1979. London: 91-100.
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.
19 Antovic A. The Overall Haemostasis Potential (OHP): a laboratory tool for the investigation of global haemostasis.. Semin Thromb Haemost 2010; 36: 772-779.
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.
21 Hoffman M. A cell-based model of coagulation and the role of factor VIIa.. Blood Rev 2003; 17 (Suppl. 01) S1-5.
22 Blomback B, Hessel B, Hogg D. et al. A two-step fibrinogen--fibrin transition in blood coagulation.. Nature 1978; 275: 501-505.
23 Hantgan RR, Hermans J. Assembly of fibrin. A light scattering study.. J Biol Chem 1979; 254: 11272-11281.