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Am J Physiol Heart Circ Physiol 292: H1876-H1882, 2007. First published December 15, 2006; doi:10.1152/ajpheart.00708.2006
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Real-time imaging of mechanically injured femoral artery in mice reveals a biphasic pattern of leukocyte accumulation

¹Life Science and Bioethics Research Center, Tokyo Medical and Dental University, and ²Department of Biochemistry and Integrative Medical Biology, School of Medicine, Keio University, Tokyo, Japan

Submitted 4 July 2006 ; accepted in final form 7 December 2006

Wire injury of an artery has been recognized as a standard model of vascular inflammation and atherosclerosis; however, the mechanism of leukocyte recruitment has not been studied in this model. In this study, we documented the recruitment of leukocytes to the murine femoral artery after a wire injury. A transluminal mechanical injury was generated by insertion of a wire into the femoral artery of male C57BL/6J mice. The mice were anesthetized and ventilated after tracheotomy and protected from hypothermia by a warming lamp. Body temperature and blood pH did not significantly change during the experiment. The interaction between rhodamine 6G-labeled leukocytes and the injured femoral artery was monitored using an epifluorescent microscope, and the images were evaluated using a computer-assisted image analysis program. In the absence of injury, virtually no leukocyte adhesion was observed. In contrast, the number of adherent leukocytes increased 4 and 24 h after injury and declined 72 h after injury. The rolling flux of leukocytes increased 4 h after injury and remained high up to 7 days, but it was faster 72 h after injury. We identified another peak of leukocyte adhesion 7 days after injury. Injection of anti-P-selectin antibody significantly reduced leukocyte adhesion at the early and later phases. In conclusion, we have established a novel experimental system for direct observation of leukocyte recruitment to the injured femoral artery. Our system revealed a previously undetected, unique profile of leukocyte recruitment during vascular injury.

vascular injury; leukocyte adhesion; intravital microscopy; inflammation

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