The overall goal of this procedure is to implant a stent into the mouse carotid artery that allows for the study of vessel remodeling or the effects of different drug coatings. First, the stent is prepared for implantation by transferring it into a silicone tube. Next, the common artery is isolated and sutures are put in place to interrupt the blood flow.
The stent is then introduced through a small incision on the external carotid artery. Once in place, the silicone tube and guide wire are removed, leaving the stent in the common carotid artery. Ultimately, samples are stained with gemsa for the analysis of plaque formation.
Existing model for instant restenosis in misa difficult, they require precise surgical skills and common complication and splitting or paralyzes. The risk is complication is minimizing our model by keeping the stent in view throughout the implantation procedure demonstrated. Parts of this procedure will be Dr.Simsek Imas postdoc from our laboratory father.
I will demonstrate the implantation of the stent into the cartridge artery. Begin by using forceps to transfer the stent to a two centimeter silicone tube. Stopping two millimeters short of the front end, the exposed end of the tube should be cut diagonally for easier implantation moisten the stent before implantation to make insertion easier.
Next, confirm the proper depth of anesthesia in an adult male mouse. After applying protective ointment to the eyes, shave the ventral neck area. Next, clean the site with repeated rubbings of Betadine and alcohol working under a stereo microscope.
Use scissors to make a small median incision over the area. Separate through the fat and tissue until the left common carotid artery is isolated. Once visualized, tie a suture around the common artery, approximately one centimeter from the bifurcation.
Tie two sutures around the external carotid artery, leaving the distal knot loose to age in stent implantation. Next, tie a suture around the internal carotid artery. After the sutures are secured, make a small incision in the external artery near the proximal knot.
Using a guide wire, introduce the silicone tube containing the stent. Once in place, remove the silicone tube to allow the stent to expand. Next, tighten the distal knot on the external carotid artery and remove the sutures on the internal and common carotid artery to restore blood flow.
Close the incision with suture clips and place the animal under a warming lamp. Until fully recovered. Word plaque formation is examined at different time points after stent implantation.
First after euthanasia and trans cardial perfusion, isolate the left carotid artery and remove the portion containing the stent. Place the sample into 4%PFA solution. At least 16 hours later, embed the sample in plastic as shown here.
After cutting 50 micrometer thick sections with a diamond band saw performed gem sustaining to evaluate plaque formation. Microcomputer tomography imaging performed one week after stent implantation showed that the stent was not dislocated by blood flow. No vessel or endothelial damage was found in untented areas of the vessel, as shown by Odin blue and endothelial specific CD 31 staining histological staining is used to study the extent of plaque formation.
Here the stent is visible as the black stained fibrin inside the neointima. In some cases, a complete occlusion of the vessel is observed. The reendothelialization of the stent struts is shown on the left by the double arrow, while the incomplete luminal reendothelialization is shown on the right by a single arrow.
While attempting this procedure, it is important to remember that the operating area should be constantly watered to prevent drying and to make manipulating the stent easier.
