title: The effect of platelets on fibrin gel structure formed in the presence of recombinant factor VIIa in hemophilia plasma and in plasma from a patient with Glanzmann thrombasthenia.
abstract: Fibrin gel structure has been shown to be dependent on the thrombin concentration as well as the rate of thrombin generation. Accordingly, factor VIII (FVIII)- and FIX-deficient plasma (hemophilia A and B) form loose fibrin clots with high permeability constants. By adding rFVIIa in vitro to FVIII-deficient plasma containing platelets (frozen and thawed), the fibrin gel permeability constant normalized, indicating that extra rFVIIa (1.2 microg mL(-1) or higher) induced a tight fibrin structure. Thrombin generation is highly dependent on the number of platelets, and in this study it was demonstrated that the addition of rFVIIa (5 microg mL(-1)) normalizes the fibrin gel permeability in samples containing platelets (frozen-thawed) in numbers of at least down to 20 x 10(6) mL(-1). The effect of rFVIIa was not observed when unfrozen platelets instead of frozen-thawed platelets were added. Neither was any effect on the fibrin permeability seen, in the presence of annexin V, known to block the effect of phospholipids on the platelet surface. This indicates an important role of platelet phospholipids for the effect of rFVIIa. A similar effect on the fibrin permeability of rFVIIa was observed when added to platelet-rich plasma from a patient with Glanzmann thrombasthenia. Recombinant FVIIa has been found to induce hemostasis in patients with hemophilia and inhibitors against FVIII/FIX as well as in patients with Glanzmann thrombasthenia, indicating the importance of the formation of a tight fibrin gel structure, more resistant against premature proteolysis, for maintaining hemostasis. In conclusion, the addition of rFVIIa (5 microg mL(-1)) also substantially decreased the permeability constant of fibrin gels formed in FVIII-deficient plasma in the presence of low numbers of frozen-thawed platelets (down to 20 x 10(6) mL(-1)). A similar pattern was obtained in plasma from a Glanzmann patient. No effect was found in the presence of unfrozen instead of frozen-thawed platelets. Annexin V blocked any effect of rFVIIa. A normalization of the overall fibrinolysis potential (OFP) during the same condition supports the effect of rFVIIa on the fibrin permeability in the presence of a limited number of platelets.
authors.affiliation: Department of Surgical Sciences, Karolinska University Hospital and Karolinska Institutet, Stockholm, Sweden. UHe@novonordisk.com
authors.forename: S
authors.initials: S
authors.lastname: He
authors.forename: G Jacobsson
authors.initials: GJ
authors.lastname: Ekman
authors.forename: U
authors.initials: U
authors.lastname: Hedner
mesh_terms: D000328:Adult; D017304:Annexin A5 / Q000494:pharmacology; D001792:Blood Platelets / Q000502:physiology*; D015925:Cryopreservation; D005167:Factor VII / Q000494:pharmacology*; D015942:Factor VIIa; D005260:Female; D005337:Fibrin / Q000737:chemistry*; D005782:Gels; D006467:Hemophilia A / Q000097:blood*; D006487:Hemostasis / Q000187:drug effects; D006801:Humans; D018613:Microscopy, Confocal; D008875:Middle Aged; D010539:Permeability; D010743:Phospholipids / Q000502:physiology; D011994:Recombinant Proteins / Q000494:pharmacology*; D013915:Thrombasthenia / Q000097:blood*
chemical_list: D017304:Annexin A5; D005782:Gels; D010743:Phospholipids; D011994:Recombinant Proteins; D005167:Factor VII; D005337:Fibrin; C103587:recombinant FVIIa; D015942:Factor VIIa