Balloon Injury of Rat Left Carotid Artery

All procedures were conducted in compliance with both the Guide for the Animal Care and Use Committee of Xinhua Hospital at SJTUSM and the guidance for the care and use of experimental animals published by NIH (the eighth Edition, NRC 2011). The experiments were approved by the Animal Care and Use Committee of Xinhua Hospital at SJTUSM. Male Sprague-Dawley rats in 10~12 weeks-old (300~350g) were anesthetized and heparinized by intraperitoneal injections of 2% Phenobarbital (40 mg/kg) and heparin (100~200 U/kg). After the bifurcation of LCA was surgically exposed, a 2-F Fogarty balloon catheter (diameter of balloon 2 mm, length 15 mm, Medtronic Company, USA) was inserted into LCA up to the aortic arch bifurcation from the external carotid artery. A pressure about 0.5–0.7 atm was given to inflate balloon and the same segment of common carotid artery was injured three times. After ligation of the external carotid artery, blood flow between the common and the internal carotid artery was confirmed to be recovered. For some of the rats, the injured carotids were surrounded by 300 μL of 30% Pluronic F-127 gel (Sigma) with or without 200ug/mL FGF10 or 10μM siFGF10 before closure of skin. All rats were housed in the Xinhua Hospital animal facilities. Four, seven, and fourteen days later, the injured carotids were excised for analyses.