Download PDF
Thromb Haemost 2014; 112(02): 412-418
DOI: 10.1160/TH13-11-0919
Animal Models
Schattauer GmbH

Comparison of the effect of coagulation and platelet function impairments on various mouse bleeding models

Nima Vaezzadeh
1   Thrombosis and Atherosclerosis Research Institute, McMaster University, Hamilton, Canada
2   Department of Medical Sciences, McMaster University, Hamilton, Canada
,
Ran Ni
1   Thrombosis and Atherosclerosis Research Institute, McMaster University, Hamilton, Canada
2   Department of Medical Sciences, McMaster University, Hamilton, Canada
,
Paul Y. Kim
1   Thrombosis and Atherosclerosis Research Institute, McMaster University, Hamilton, Canada
3   Department of Medicine, McMaster University, Hamilton, Canada
,
Jeffrey I. Weitz
1   Thrombosis and Atherosclerosis Research Institute, McMaster University, Hamilton, Canada
2   Department of Medical Sciences, McMaster University, Hamilton, Canada
3   Department of Medicine, McMaster University, Hamilton, Canada
4   Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Canada
,
Peter L. Gross
1   Thrombosis and Atherosclerosis Research Institute, McMaster University, Hamilton, Canada
2   Department of Medical Sciences, McMaster University, Hamilton, Canada
3   Department of Medicine, McMaster University, Hamilton, Canada
› Author Affiliations Financial support: This study was supported by a Canadian Blood Services/ Canadian Institutes of Health Research Operating Grant and an Ontario Graduate Scholarship.
Further Information

Publication History

Received: 07 November 2013

Accepted after major revision: 08 March 2014

Publication Date:
04 December 2017 (online)

    Permissions and Reprints

Summary

Haemostatic impairments are studied in vivo using one of several murine bleeding models. However it is not known whether these models are equally appropriate for assessing coagulation or platelet function defects. It was our study objective to assess the performance of arterial, venous and combined arterial and venous murine bleeding models towards impaired coagulation or platelet function. Unfractionated heparin (UFH) or αIIbβ3 inhibitory antibody (Leo.H4) were administered to mice, and their effects on bleeding in saphenous vein, artery, and tail tip transection models were quantified and correlated with their effects on plasma clotting and ADP-induced platelet aggregation, respectively. All models exhibited similar sensitivity with UFH (EC50 dose = 0.19, 0.13 and 0.07 U/g, respectively) (95% CI = 0.14 – 0.27, 0.08 – 0.20, and 0.03 – 0.16 U/g, respectively). Maximal inhibition of ex vivo plasma clotting could be achieved with UFH doses as low as 0.03 U/g. In contrast, the saphenous vein bleeding model was less sensitive to αIIbβ3 inhibition (EC50 = 6.9 µg/ml) than tail transection or saphenous artery bleeding models (EC50 = 0.12 and 0.37 µg/ml, respectively) (95% CI = 2.4 – 20, 0.05 – 0.33, and 0.06 – 2.2 µg/ml, respectively). The EC50 of Leo.H4 for ADP-induced platelet aggregation in vitro (8.0 µg/ml) was at least 20-fold higher than that of the tail and arterial, but not the venous bleeding model. In conclusion, venous, arterial and tail bleeding models are similarly affected by impaired coagulation, while platelet function defects have a greater influence in models incorporating arterial injury.

Keywords

Haemorrhage - experimental animal models - heparin - platelet aggregation inhibitors
 

  • References

  • 1 White TA, Johnson T, Zarzhevsky N. et al. Endothelial-derived tissue factor pathway inhibitor regulates arterial thrombosis but is not required for development or hemostasis. Blood 2010; 116: 1787-1794.
    CrossrefPubMedGoogle Scholar
  • 2 Stolla M, Stefanini L, Roden RC. et al. The kinetics of alphaIIbbeta3 activation determines the size and stability of thrombi in mice: implications for antiplatelet therapy. Blood 2011; 117: 1005-1013.
    CrossrefPubMedGoogle Scholar
  • 3 Petrich BG, Marchese P, Ruggeri ZM. et al. Talin is required for integrin-mediated platelet function in hemostasis and thrombosis. J Exp Med 2007; 204: 3103-3111.
    CrossrefPubMedGoogle Scholar
  • 4 Andre P, Delaney SM, LaRocca T. et al. P2Y12 regulates platelet adhesion/activation, thrombus growth, and thrombus stability in injured arteries. J Clin Invest 2003; 112: 398-406.
    CrossrefPubMedGoogle Scholar
  • 5 Sheffield WP, Eltringham-Smith LJ, Gataiance S. et al. A long-lasting, plasmin-activatable thrombin inhibitor aids clot lysis in vitro and does not promote bleeding in vivo. Thromb Haemost 2009; 101: 867-877.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 6 Cho J, Furie BC, Coughlin SR. et al. A critical role for extracellular protein disulfide isomerase during thrombus formation in mice. J Clin Invest 2008; 118: 1123-1131.
    PubMedGoogle Scholar
  • 7 Rand ML, Wang H, Pluthero FG. et al. Diannexin, an annexin A5 homodimer, binds phosphatidylserine with high affinity and is a potent inhibitor of platelet-mediated events during thrombus formation. J Thromb Haemost 2012; 10: 1109-1119.
    CrossrefPubMedGoogle Scholar
  • 8 Matsuno H, Kozawa O, Okada K. et al. Plasmin generation plays different roles in the formation and removal of arterial and venous thrombus in mice. Thromb Haemost 2002; 87: 98-104.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 9 Bird JE, Wang X, Smith PL. et al. A platelet target for venous thrombosis? P2Y1 deletion or antagonism protects mice from vena cava thrombosis. J Thromb Thrombolysis 2012; 34: 199-207.
    CrossrefPubMedGoogle Scholar
  • 10 Schumacher WA, Bostwick JS, Ogletree ML. et al. Biomarker optimization to track the antithrombotic and hemostatic effects of clopidogrel in rats. J Pharmacol Exp Ther 2007; 322: 369-377.
    CrossrefPubMedGoogle Scholar
  • 11 Savage B, Almus-Jacobs F, Ruggeri ZM. Specific synergy of multiple substrate-receptor interactions in platelet thrombus formation under flow. Cell 1998; 94: 657-666.
    CrossrefPubMedGoogle Scholar
  • 12 Nesbitt WS, Westein E, Tovar-Lopez FJ. et al. A shear gradient-dependent platelet aggregation mechanism drives thrombus formation. Nat Med 2009; 15: 665-673.
    CrossrefPubMedGoogle Scholar
  • 13 Ruggeri ZM, Orje JN, Habermann R. et al. Activation-independent platelet adhesion and aggregation under elevated shear stress. Blood 2006; 108: 1903-1910.
    CrossrefPubMedGoogle Scholar
  • 14 Maxwell MJ, Westein E, Nesbitt WS. et al. Identification of a 2-stage platelet aggregation process mediating shear-dependent thrombus formation. Blood 2007; 109: 566-576.
    CrossrefPubMedGoogle Scholar
  • 15 Repke D, Gemmell CH, Guha A. et al. Hemophilia as a defect of the tissue factor pathway of blood coagulation: effect of factors VIII and IX on factor X activation in a continuous-flow reactor. Proc Natl Acad Sci USA 1990; 87: 7623-7627.
    CrossrefPubMedGoogle Scholar
  • 16 Neeves KB, Illing DA, Diamond SL. Thrombin flux and wall shear rate regulate fibrin fiber deposition state during polymerization under flow. Biophys J 2010; 98: 1344-1352.
    CrossrefPubMedGoogle Scholar
  • 17 Campbell RA, Aleman M, Gray LD. et al. Flow profoundly influences fibrin network structure: implications for fibrin formation and clot stability in haemostasis. Thromb Haemost 2010; 104: 1281-1284.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 18 Duke WW. The relation of blood platelets to hemorrhagic disease: description of a method for determining the bleeding time and coagulation time and report of three cases of hemorrhagic disease relieved by transfusion. J Am Med Assoc 1910; 55: 1185-1192.
    CrossrefPubMedGoogle Scholar
  • 19 George JN, Caen JP, Nurden AT. Glanzmann’s thrombasthenia: the spectrum of clinical disease. Blood 1990; 75: 1383-1395.
    CrossrefPubMedGoogle Scholar
  • 20 George JN. Platelets. Lancet 2000; 355: 1531-1539.
    CrossrefPubMedGoogle Scholar
  • 21 Curtis BR, Swyers J, Divgi A. et al. Thrombocytopenia after second exposure to abciximab is caused by antibodies that recognize abciximab-coated platelets. Blood 2002; 99: 2054-2059.
    CrossrefPubMedGoogle Scholar
  • 22 Simpfendorfer C, Kottke-Marchant K, Lowrie M. et al. First chronic platelet glycoprotein IIb/IIIa integrin blockade. A randomized, placebo-controlled pilot study of xemilofiban in unstable angina with percutaneous coronary interventions. Circulation 1997; 96: 76-81.
    CrossrefPubMedGoogle Scholar
  • 23 Peyvandi F, Mannucci PM. Rare coagulation disorders. Thromb Haemost 1999; 82: 1207-1214.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 24 Borchgrevink CF, WAALER BA. The secondary bleeding time; a new method for the differentiation of hemorrhagic diseases. Acta Med Scand 1958; 162: 361-374.
    CrossrefPubMedGoogle Scholar
  • 25 Borchgrevink CF. Platelet adhesion in vivo in patients with bleeding disorders. Acta Med Scand 1961; 170: 231-243.
    CrossrefPubMedGoogle Scholar
  • 26 Kaneshiro MM, Mielke Jr CH, Kasper CK. et al. Bleeding time after aspirin in disorders of intrinsic clotting. N Engl J Med 1969; 281: 1039-1042.
    CrossrefPubMedGoogle Scholar
  • 27 Buyue Y, Whinna HC, Sheehan JP. The heparin-binding exosite of factor IXa is a critical regulator of plasma thrombin generation and venous thrombosis. Blood 2008; 112: 3234-3241.
    CrossrefPubMedGoogle Scholar
  • 28 Pastoft AE, Lykkesfeldt J, Ezban M. et al. A sensitive venous bleeding model in haemophilia A mice: effects of two recombinant FVIII products (N8 and Advate((R))). Haemophilia 2012; 18: 782-788.
    CrossrefPubMedGoogle Scholar
  • 29 Kim PY, Stewart RJ, Lipson SM. et al. The relative kinetics of clotting and lysis provide a biochemical rationale for the correlation between elevated fibrinogen and cardiovascular disease. J Thromb Haemost 2007; 05: 1250-1256.
    CrossrefPubMedGoogle Scholar
  • 30 Wang X, Xu L. An optimized murine model of ferric chloride-induced arterial thrombosis for thrombosis research. Thromb Res 2005; 115: 95-100.
    CrossrefPubMedGoogle Scholar
  • 31 Lavelle SM, MacIomhair M. Bleeding times and the antithrombotic effects of high-dose aspirin, hirudin and heparins in the rat. Ir J Med Sci 1998; 167: 216-220.
    CrossrefPubMedGoogle Scholar
  • 32 Lambourne MD, Eltringham-Smith LJ, Gataiance S. et al. Prothrombin complex concentrates reduce blood loss in murine coagulopathy induced by warfarin, but not in that induced by dabigatran etexilate. J Thromb Haemost 2012; 10: 1830-1840.
    CrossrefPubMedGoogle Scholar
  • 33 Li X, Cooley BC. Effect of anticoagulation and inhibition of platelet aggregation on arterial versus venous microvascular thrombosis. Ann Plast Surg 1995; 35: 165-169.
    CrossrefPubMedGoogle Scholar
  • 34 Hupkens P, Cooley BC. Comparison of arterial and venous patency in a rat model of subendothelium-stimulated thrombosis. Microsurgery 1996; 17: 226-229.
    CrossrefPubMedGoogle Scholar
  • 35 Aird WC. Vascular bed-specific thrombosis. J Thromb Haemost 2007; 05 (Suppl. 01) 283-291.
    CrossrefPubMedGoogle Scholar
  • 36 Yamamoto H, Vreys I, Stassen JM. et al. Antagonism of vWF inhibits both injury induced arterial and venous thrombosis in the hamster. Thromb Haemost 1998; 79: 202-210.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 37 Baumgartner HR. The role of blood flow in platelet adhesion, fibrin deposition, and formation of mural thrombi. Microvasc Res 1973; 05: 167-179.
    CrossrefPubMedGoogle Scholar
  • 38 Turitto VT, Baumgartner HR. Platelet interaction with subendothelium in flowing rabbit blood: effect of blood shear rate. Microvasc Res 1979; 17: 38-54.
    CrossrefPubMedGoogle Scholar
  • 39 Turitto VT, Weiss HJ, Baumgartner HR. The effect of shear rate on platelet interaction with subendothelium exposed to citrated human blood. Microvasc Res 1980; 19: 352-365.
    CrossrefPubMedGoogle Scholar
  • 40 Weiss EJ, Hamilton JR, Lease KE. et al. Protection against thrombosis in mice lacking PAR3. Blood 2002; 100: 3240-3244.
    CrossrefPubMedGoogle Scholar
  • 41 Kahn ML, Zheng YW, Huang W. et al. A dual thrombin receptor system for platelet activation. Nature 1998; 394: 690-694.
    CrossrefPubMedGoogle Scholar
  • 42 Maroney SA, Cooley BC, Ferrel JP. et al. Absence of hematopoietic tissue factor pathway inhibitor mitigates bleeding in mice with hemophilia. Proc Natl Acad Sci USA 2012; 109: 3927-3931.
    CrossrefPubMedGoogle Scholar
  • 43 Broze Jr GJ, Yin ZF, Lasky N. A tail vein bleeding time model and delayed bleeding in hemophiliac mice. Thromb Haemost 2001; 85: 747-748.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 44 Liu Y, Jennings NL, Dart AM. et al. Standardizing a simpler, more sensitive and accurate tail bleeding assay in mice. World J Exp Med 2013; 02: 30-36.
    CrossrefPubMedGoogle Scholar
  • 45 Hashimoto M, Sugidachi A, Isobe T. et al. The influence of P2Y12 receptor deficiency on the platelet inhibitory activities of prasugrel in a mouse model: evidence for specific inhibition of P2Y12 receptors by prasugrel. Biochem Pharmacol 2007; 74: 1003-1009.
    CrossrefPubMedGoogle Scholar
  • 46 Gregersen MI, Rawson RA. Blood volume. Physiol Rev 1959; 39: 307-342.
    CrossrefPubMedGoogle Scholar
  • 47 Suh TT, Holmback K, Jensen NJ. et al. Resolution of spontaneous bleeding events but failure of pregnancy in fibrinogen-deficient mice. Genes Dev 1995; 09: 2020-2033.
    CrossrefPubMedGoogle Scholar