Mechanisms underlying FeCl3-induced arterial thrombosis

UMR-S949 Inserm-Université de Strasbourg, Strasbourg, France.
Journal of Thrombosis and Haemostasis (Impact Factor: 5.55). 04/2011; 9(4):779-89. DOI: 10.1111/j.1538-7836.2011.04218.x
Source: PubMed

ABSTRACT The FeCl(3)-induced vascular injury model is widely used to study thrombogenesis in vivo, but the processes leading to vascular injury and thrombosis are poorly defined.
The aim of our study was to better characterize the mechanisms of FeCl(3)-induced vascular injury and thrombus formation, in order to evaluate the pathophysiological relevance of this model.
FeCl(3) was applied at different concentrations (from 7.5% to 20%) and for different time periods (up to 5 min) to mouse carotid or mesenteric arteries.
Under all the conditions tested, ultrastructural analysis revealed that FeCl(3) diffused through the vessel wall, resulting in endothelial cell denudation without exposure of the inner layers. Hence, only the basement membrane components were exposed to circulating blood cells and might have contributed to thrombus formation. Shortly after FeCl(3) application, numerous ferric ion-filled spherical bodies appeared on the endothelial cells. Interestingly, platelets could adhere to these spheres and form aggregates. Immunogold labeling revealed important amounts of tissue factor at their surface, suggesting that these spheres may play a role in thrombin generation. In vitro experiments indicated that FeCl(3) altered the ability of adhesive proteins, including collagen, fibrinogen and von Willebrand factor, to support platelet adhesion. Finally, real-time intravital microscopy showed no protection against thrombosis in GPVI-immunodepleted and β(1)(-/-) mice, suggesting that GPVI and β(1) integrins, known to be involved in initial platelet adhesion and activation, do not play a critical role in FeCl(3)-induced thrombus formation.
This model should be used cautiously, in particular to study the earliest stage of thrombus formation.

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    • "In this model FeCl 3 is applied to the topical surface of an intact vessel, triggering vascular wall injury and denudation of the endothelium via a mechanism involving the generation of reactive oxygen species [4] [14]. A recent study suggested that FeCl 3 -induced vascular injury was erythrocyte-dependent , requiring hemolysis and hemoglobin oxidation for endothelial denudation [20] [21]. One measurable parameter in this model is the elapsed time from injury to complete vessel occlusion, measured as blood flow cessation by Doppler flowmeter or under direct observation with intravital microscopy [13] [14] [15]. "
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    ABSTRACT: Ferric chloride (FeCl3) induced vascular injury is a widely used model of occlusive thrombosis that reports platelet activation in the context of an aseptic closed vascular system. This model is based on redox-induced endothelial cell injury, which is simple and sensitive to both anticoagulant and anti-platelets drugs. The time required for platelet aggregation to occlude blood flow gives a quantitative measure of vascular damage that is pathologically relevant to thrombotic disease. We have refined the traditional FeCl3-induced carotid artery model making the data highly reproducible with lower variation. This paper will describe our artifices and report the role of varying the oxidative damage by varying FeCl3 concentrations and exposure. To explore a maximum difference between experimental groups, adjustment of the selected FeCl3 dose and exposure duration may be necessary.
    01/2013; 1(1):50-5. DOI:10.1016/j.redox.2012.11.001
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    • "destruction of endothelial cells and results in occlusive thrombus formation (Eckly et al., 2011; Kurz, Main, & Sandusky, 1990; Tseng, Dozier, Haribabu, & Graham, 2006). Shortly after FeCl 3 application, numerous ferric ion-filled spherical bodies appeared to be budding off the endothelial cells into the lumen, on which platelets could adhere and form aggregates to generate a thrombus (Eckly et al., 2011). The FeCl 3 -induced arterial model of thrombosis is a wellestablished model particularly in mice and rats (Couture, Richer, Cadieux, Thomson, & Hossain, 2011; Heran et al., 2000; Kurz et al., 1990; Leadley, Chi, Rebello, & Gagnon, 2000; Surin, Prakash, Barthwal, & Dikshit, 2010; Wang & Xu, 2005). "
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    ABSTRACT: INTRODUCTION: The FeCl(3)-induced arterial model of thrombosis is one of the most widely used animal models to assess arterial efficacy of new antithrombotic drug candidates. This model is well-established in rodents but in a less extent in the rabbit. In this work, we present a methodology for a rabbit FeCl(3)-induced arterial model of thrombosis derived from our troubleshooting which allows the generation of reliable efficacy data for new antithrombotic drug candidates METHODS: Rabbits were administered with heparin 4.5 U/kg/min, argatroban 10μg/kg/min or saline by intravenous infusion. The blood flow was monitored using a Doppler flow probe. The time from the application of FeCl(3) to the recorded zero blood flow was defined as the time to occlusion, with a maximum recording time of 60min post-FeCl(3) application. After 30min of infusion, thrombosis was induced by wrapping a FeCl(3)-saturated filter paper around the carotid artery caudal to the flow probe. Animals were subject to exclusion criteria based on the visual aspect of the artery FeCl(3)-induced injury and based on changes in blood flow upon FeCl(3) application RESULTS: Following the application of FeCl(3), a mean time to occlusion for saline, heparin and argatroban of 24.3±1.8, 52.5±4.8 and 53.5±4.5min was obtained, respectively. Mean time to occlusion for heparin and argatroban administered groups were significantly different when compared to the saline-treated group (p<0.05). These results for the test compounds represent approximately 80% of the maximum possible prolongation DISCUSSION: The rabbit FeCl(3)-induced arterial model of thrombosis presented in this paper derived from our troubleshooting is sensitive and reproducible for the generation of accurate and reliable efficacy data in the assessment of new antithrombotic agents in preclinical drug development.
    Journal of pharmacological and toxicological methods 12/2012; DOI:10.1016/j.vascn.2012.11.003 · 2.15 Impact Factor
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    ABSTRACT: See also Eckly A, Hechler B, Freund M, Zerr M, Cazenave JP, Lanza F, Mangin PH, Gachet C. Mechanisms underlying FeCl3-induced arterial thrombosis. This issue, pp 779-89.
    Journal of Thrombosis and Haemostasis 02/2011; 9(4):776-8. DOI:10.1111/j.1538-7836.2011.04238.x · 5.55 Impact Factor
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