A Comparison of the in Vitro and in Vivo Thrombogenic Activity of Factor IX Concentrates Using Stasis (Wessler) and Non-Stasis Rabbit Models

Summary

The thrombogenic effects of selected factor IX concentrates were evaluated in two rabbit models; the Wessler stasis model and a novel non-stasis model. Concentrates active in either the NAPTT or TGt₅₀ in vitro tests of potential thrombogenicity, or both, caused thrombus formation in the Wessler technique and activation of the coagulation system in the non-stasis model. A concentrate with low activity in both in vitro tests did not have thrombogenic effects in vivo, at the chosen dose. Results in the non-stasis model suggested that the thrombogenic effects of factor IX concentrates may occur by at least two mechanisms. A concentrate prepared from platelet-rich plasma and a pyrogenic concentrate were also tested and found to have no thrombogenic effect in vivo.

These studies justify the use of the NAPTT and TGt₅₀ in vitro tests for the screening of factor IX concentrates prior to clinical use.

Key words

Factor IX concentrate - Thrombosis - Intravascular coagulation

• References

• 1 Menaché D, Fauvert R, Soulier JP. Utilisation en hépatologie d'une fractione contenant la prothrombine, le complexe proconvertine – Stuart et le facteur hémophilique B. Pathol Biol (Paris) 1959; 7: 2515-2523
• 2 Tullis JL, Breen F. Christmas factor concentrates: the clinical use of several preparations. Bibl Haematol 1970; 34: 40-51
• 3 Kasper CK. Thromboembolic complications (following the use of factor IX concentrates). Thromb Diath Haemorrh 1975; 33: 640-644
• 4 Cash JD, Dalton RG, Middleton S, Smith JK. Studies on the thrombogenicity of Scottish factor IX concentrates in dogs. Thromb Diath Haemorrh 1975; 33: 632-639
• 5 Hedner U, Nilsson IM, Bergentz SE. Various prothrombin complex concentrates and their effect on coagulation and fibrinolysis in vivo. Thromb Haemostas 1976; 35: 386-395
• 6 Hedner U, Nilsson IM, Bergentz SE. Studies on the thrombogenic activities in two prothrombin complex concentrates. Thromb Haemostas 1979; 42: 1022-1032
• 7 Preston FE, Winfield DA, Malia RG, Blackburn EK. Serial coagulation changes in the coagulation system following clotting factor concentrate infusion. Thromb Diath Haemorrh 1975; 34: 475-482
• 8 Preston FE, Malia RG, Lilleyman JS, Blackburn EK. Heparinised clotting factor concentrates in patients with Christmas disease and liver disease. Thromb Haemostas 1977; 38: 504-509
• 9 Kingdon HS, Lundblad RL, Veltkamp JJ, Aronson DL. Potentially thrombogenic materials in factor IX concentrates. Thromb Diath Haemorrh 1975; 33: 617-631
• 10 Sas G, Owens RE, Smith JK, Middleton S, Cash JD. In vitro spontaneous thrombin generation in human factor IX concentrates. Br J Haematol 1975; 31: 25-35
• 11 Cash JD, Owens R, Dalton RG, Prescott RJ. Thrombogenicity of factor IX concentrates: in vitro and in vivo (rabbit) studies. Vox Sang 1978; 35: 105-110
• 12 Wessler S, Reimer SM, Sheps MC. Biologic assay of a thrombosis-inducing activity in human serum. J Appl Physiol 1959; 14: 943-946
• 13 Pepper DS, Banheygi D, Howie A, Cash JD. In vitro thrombogenicity tests of factor IX concentrates. Br J Haematol 1977; 36: 573-583
• 14 Prowse CV, Pepper DS, Cash JD, Patterson M. Thrombogenicity screening of factor IX concentrates. Thromb Haemostas 1977; 38: 728-729
• 15 Prowse CV, Chirnside A, Elton R. In vitro thrombogenicity tests of factor IX concentrates. I: A survey of available assays. Thromb Haemostas 1979; 42: 1355-1367
• 16 Foster PR, Patterson MR, Johnson AJ, Middleton SM. Thrombogenicity of factor IX concentrates and polyethylene glycol processing. Thromb Res 1980; 17: 273-279
• 17 Pepper DS, Prowse CV. Chromatography of human prothrombin complex on dextran sulphate agarose. Thromb Res 1977; 11: 687-692
• 18 Levin J, Bang F. Clottable protein in Limulus blood: its localisation and kinetics of its coagulation by endotoxin. Fed Proc 1966; 25: 497
• 19 Ellis BC, Stransky A. A quick and accurate method for the determination of fibrinogen in plasma. J Lab Clin Med 1961; 58: 477-488
• 20 Hardisty RM, Ingram GI C. Bleeding Disorders: Investigation and management. Blackwell Scientific Publications Ltd., Oxford: 1965
• 21 Kierulf P, Godal H. Fibrinemia in medical patients screened by the ethanol gelation test. Acta Med Scand 1971; 190: 185-190
• 22 Smith JK, Snape TJ, Bidwell E, Edwards R, Gunson HH. Preparation of factor IX concentrate from platelet-rich plasma. Thromb Haemostas 1977; 37: 363-366
• 23 Prowse CV, Boffa MC, Guthrie C, Pepper DS. In vitro thrombogenicity tests of factor IX concentrates. II: Effects of phospholipids and heparin. Thromb Haemostas 1979; 42: 1368-1377
• 24 Wessler S, Yin ET. Experimental hypercoagulable state induced by factor X. Comparison of the non-activated and activated forms. J Lab Clin Med 1968; 72: 256-260
• 25 Triantaphyllopoulous DC. Intravascular coagulation following injection of prothrombin complex. Am J Clin Pathol 1972; 57: 603-610
• 26 Vermylen J, Schetz J, Semeraro N, Mertens F, Verstraete M. Evidence that activated prothrombin concentrates enhance platelet coagulant activity. Br J Haematol 1978; 38: 235-242
• 27 Penner J, Leach K, Rohwedder J. Thrombogenic characteristics of activated and standard concentrates of prothrombin complex. Thromb Haemostas 1977; 38: 316a
• 28 Giles AR, Mann KG, Nesheim ME, Tracy PB, Blajchman MA. A correlation between the coagulant active phospholipid content of prothrombin complex concentrates and in vivo thrombogenicity. Blood 1979; 54: 279 a