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Thromb Haemost 1997; 78(05): 1408-1414
DOI: 10.1055/s-0038-1665420
Review Article
Schattauer GmbH Stuttgart

A New Model for Quantitative In Vivo Microscopic Analysis of Thrombus Formation and Vascular Recanalisation: The Ear of the Hairless (hr/hr) Mouse

Frank Roesken
1   The institute for Clinical & Experimental Surgery, University of Saarland, Homburg/Saar, Germany
,
Martin Ruecker
1   The institute for Clinical & Experimental Surgery, University of Saarland, Homburg/Saar, Germany
,
Brigitte Vollmar
1   The institute for Clinical & Experimental Surgery, University of Saarland, Homburg/Saar, Germany
,
Nicole Boeckel
2   institute for Medical Biology, University of Saarland, Homburg/Saar, Germany
,
Eberhard Morgenstern
2   institute for Medical Biology, University of Saarland, Homburg/Saar, Germany
,
Michael D Menger
1   The institute for Clinical & Experimental Surgery, University of Saarland, Homburg/Saar, Germany
› Author Affiliations
Further Information

Publication History

Received 20 1997

Accepted after revision 15 July 1997

Publication Date:
12 July 2018 (online)

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Summary

The alteration of rheological blood properties as well as deterioration of vascular perfusion conditions and cell-cell interactions are major determinants of thrombus formation. Herein, we present an experimental model which allows for quantitative in vivo microscopic analysis of these determinants during both thrombus formation and vascular recanalisation. The model does not require surgical preparation procedures, and enables for repeated analysis of identical microvessels over time periods of days or months, respectively. After i.v. administration of FITC-dextran thrombus formation was induced photochemically by light exposure to individual arterioles and venules of the ear of ten anaesthetised hairless mice. In venules, epiillumination induced rapid thrombus formation with first platelet deposition after 0.59 ± 0.04 min and complete vessel occlusion within 7.48 ±1.31 min. After a 24-h time period, 75% of the thrombosed venules were found recanalised. Marked leukocyte-endothelial cell interaction in those venules indicated persistent endothelial cell activation and/or injury, even after an observation period of 7 days. In arterioles, epi-illumination provoked vasomotion, while thrombus formation was significantly (p <0.05) delayed with first platelet deposition after 2.32 ± 0.22 min and complete vessel occlusion within 20.07 ±3.84 min. Strikingly, only one of the investigated arterioles was found recanalised after 24 h, which, however, did not show leukocyte-endothelial cell interaction. Heparin (300 U/kg, i.v.) effectively counteracted the process of thrombus formation in this model, including both first platelet deposition and vessel occlusion. We conclude that the model of the ear of the hairless mouse allows for distinct in vivo analysis of arteriolar and venular thrombus formation/ recanalisation, and, thus, represents an interesting tool for the study of novel antithrombotic and thrombolytic strategies, respectively.

 
  • 1 Aronson DL, Thomas DP. Experimental studies on venous thrombosis: effect of coagulants, procoagulants and vessel contusion. Thromb Haemost 1985; 54: 866-870
    Article in Thieme ConnectPubMedGoogle Scholar
  • 2 Baker M, Wayland H. On-line volume flow rate and velocity profile measurements for blood in microvessels. Microvasc Res 1974; 07: 131-143
    CrossrefPubMedGoogle Scholar
  • 3 Barker JH, Hammersen F, Bondar I, Uhl E, Galla TJ, Menger MD, Mess-mer K. The hairless mouse ear for in vivo studies of skin microcirculation. Plast Reconstr Surg 1989; 83: 948-959
    CrossrefPubMedGoogle Scholar
  • 4 Belch JJ. The role of the white blood cell in arterial disease. Blood Coagul Fibrinolysis 1990; 1: 183-191
    PubMedGoogle Scholar
  • 5 Bemat A, Lale A, Herbert JM. Aurin tricarboxylic acid inhibits experimental venous thrombosis. Thromb Res 1994; 74: 617-627
    CrossrefPubMedGoogle Scholar
  • 6 Chen C, Hanson SR, Lumsden AB. Boundary layer infusion of heparin prevents thrombosis and reduces neointimal hyperplasia in venous polytetra- fluoroethylene grafts without systemic anticoagulation. J Vase Surg 1995; 22: 237-247
    CrossrefPubMedGoogle Scholar
  • 7 Davidson SF, Brantley SK, Talbot PJ, Das SK. A functional model of mi- crovascular thrombosis. Plast Reconstr Surg 1990; 86: 579-581
    CrossrefPubMedGoogle Scholar
  • 8 Dodds WJ. Animal models for the evolution of thrombotic disease. Ann N Y Acad Sci 1987; 516: 631-635
    CrossrefPubMedGoogle Scholar
  • 9 Fagrell B, Eriksson SE, Malmstmm S, Sjjclund A. Computerized data analysis of capillary blood cell velocity in humans (Abstract). Int J Microcirc: Clin Exp 1988; 07: S40
    PubMedGoogle Scholar
  • 10 Fareed J, Jeske W, Eschenfelder V, Iqbal O, Hoppensteadt D, Ahsan A. Pharmacologic validation of the clinical effects of an optimized low-molec- ular-weight heparin-reviparin. Semin Thromb Hemost 1995; 21: 212-227
    Article in Thieme ConnectPubMedGoogle Scholar
  • 11 Gralnick HR, Williams S, McKeown LP, Hansmann K, Fenton JW, Krutzsch H. High-affinity alpha-thrombin binding to platelet glycoprotein lb alpha: identification of two binding domains. Proc Natl Acad Sci USA 1994; 91: 6334-6338
    CrossrefPubMedGoogle Scholar
  • 12 Haines ST, Bussey HI. Thrombosis and the pharmacology of antithrombotic agents. Ann-Pharmacother 1995; 29: 892-905
    PubMedGoogle Scholar
  • 13 Hanson SR, Kotze HF, Savage B, Harker LA. Platelet interactions with Dacron vascular grafts. A model of acute thrombosis in baboons. Arteriosclerosis 1985; 05: 595-603
    CrossrefPubMedGoogle Scholar
  • 14 Heiniger HJ, Borey JJ. In: Handbook of genetically standardized jax mice. Jackson Laboratories; Bar Harbor, USA: 1980
  • 15 Herrmann KS. Platelet aggregation induced in the hamster cheek pouch by a photochemical process with excited fluorescein isothiocyanate-dextran. Microvasc Res 1983; 26: 238-249
    CrossrefPubMedGoogle Scholar
  • 16 Hirschberg M, Hofferberth B. New model of cerebral thrombosis in dogs. Stroke 1988; 19: 741-746
    CrossrefPubMedGoogle Scholar
  • 17 Jourdan A, Aguejouf O, Imbault P, Doutremepuich F, Inamo J, Doutreme-puich C. Experimental thrombosis model induced by free radicals. Application to aspirin and other different substances. Thromb Res 1995; 79: 109-123
    CrossrefPubMedGoogle Scholar
  • 18 Kamovsky MJ. A formaldehyde-glutaraldehyde fixative of high osmolality for use in electron microscopy. J Cell Biol 1965; 27: 137-138A
    PubMedGoogle Scholar
  • 19 Laux V, Seiffge D. Platelet function in the dorsal skin fold chamber of the rat. In Vivo 1993; 7: 45-51
    PubMedGoogle Scholar
  • 20 Perry DD, Reddick RL, Risco JM. Choroidal microvascular repair after argon laser photocoagulation. Ultrastructural observations. Invest Ophthalmol Vis Sci 1984; 25: 1019-1026
    PubMedGoogle Scholar
  • 21 Piotrowicz RS, Orchekowski RP, Nugent DJ, Yamada KY, Kunicki TJ. Glycoprotein Ic-IIa functions as an activation-independent fibronectin receptor on human platelets. J Cell Biol 1988; 106: 1359-1364
    CrossrefPubMedGoogle Scholar
  • 22 Rosenblum WI. Biology of disease: Aspects of endothelial malfunction and function in cerebral microvessels. Lab Invest 1986; 55: 252-268
    PubMedGoogle Scholar
  • 23 Rote WE, Wems SW, Davis JH, Feigen LP, Kilgore KS, Lucchesi BR. Platelet GPIIb/IIIa receptor inhibition by SC-49992 prevents thrombosis and rethrombosis in the canine carotid artery. Cardiovasc Res 1993; 27: 500-507
    CrossrefPubMedGoogle Scholar
  • 24 Saniabadi AR, Umemura K, Matsumoto N, Sakuma S, Nakashima M. Vessel wall injury and arterial thrombosis induced by a photochemical reaction. Thromb Haemost 1995; 73: 868-872
    Article in Thieme ConnectPubMedGoogle Scholar
  • 25 Sato M, Ohshima N. Effect of wall shear rate on thrombogenesis in microvessels of the rat mesentery. Circ Res 1990; 66: 941-949
    CrossrefPubMedGoogle Scholar
  • 26 Seiffge D, Kremer E. Influence of ADP, blood flow velocity, and vessel diameter on the laser-induced thrombus formation. Thromb Res 1986; 42: 331-341
    CrossrefPubMedGoogle Scholar
  • 27 Siegel JM, Markou CP, Ku DN, Hanson SR. A scaling law for wall shear rate through an arterial stenosis. J Biomech Eng 1994; 116: 446-451
    CrossrefPubMedGoogle Scholar
  • 28 Stepnick DW, Hayden RE, Hogikyan ND. Development of thrombosis models in the rabbit. Arch Otolaryngol Head Neck Surg 1994; 120: 997-1002
    CrossrefPubMedGoogle Scholar
  • 29 Sugidachi A, Asai F, Tani Y, Hosokawa T, Koike H. Occlusive thrombosis in the femoral artery of the rabbit: a pharmacological model for evaluating antiplatelet and anticoagulant agents. Blood Coagul Fibrinolysis 1996; 7: 57-64
    CrossrefPubMedGoogle Scholar
  • 30 Umemura K, Kohno Y, Matsuno H, Uematsu T, Nakashima M. A new model for photochemically induced thrombosis in the inner ear microcirculation and the use of hearing loss as a measure for microcirculatory disorders. Eur Arch Otorhinolaryngol 1990; 248: 105-108
    PubMedGoogle Scholar
  • 31 van derGiessenWJ, Mooi W, Rutteman AM, van VlietHH, Slager CJ, Verdouw PD. A new model for coronary thrombosis in the pig: preliminary results with thrombolysis. Eur Heart J 1983; 04 Suppl C 69-76
    CrossrefPubMedGoogle Scholar
  • 32 Weichert W, Pauliks V, Breddin HK. Laser-induced thrombi in rat mesenteric vessels and antithrombotic drugs. Haemostasis 1983; 13: 61-71
    PubMedGoogle Scholar
  • 33 Yao SK, Akhtar S, Scott BurdenT, Ober JC, Golino P, Buja LM, Casscells W, Willerson JT. Endogenous and exogenous nitric oxide protect against intracoronary thrombosis and reocclusion after thrombolysis. Circulation 1995; 92: 1005-1010
    CrossrefPubMedGoogle Scholar
  • 34 Yokota K, Yamashita A, Oda M. Anti thrombotic activity of KBT-3022 in experimental models of thrombosis. Jpn J Pharmacol 1995; 68: 201-206
    CrossrefPubMedGoogle Scholar
  • 35 Zeintl H, Tompkins WR, Messmer K, Intaglietta M. Static and dynamic video analysis applied to clinical investigations. Prog Appl Microcir 1986; 11: 01-10
    PubMedGoogle Scholar