All experiments have been approved by the McGill University Animal Care Committee, and we're in accordance with the guidelines of the Canadian Council on Animal Care. The goal of this experiment is to deliver progenitor or stem cells into the mouse of mandibular salivary gland through the cannulation of the Wharton's duct. For this video, trippen blue solution is used as a dye to visualize the cell infusion.
Hypofunction of the salivary glands is one of the most common side effects of Sjogren's syndrome and therapeutic irradiation for cancer treatment in the head and neck regions. Malfunction of the salivary gland can often lead to dental diseases, such as infection of the oral mucosa and tooth tooth decay. One innovative approach of regenerative medicine for the treatment of damaged salivary glands is the following.
Stem cells can be directly deposited into the gland by cannulation via the ducks of the salivary gland as a potent method in reviving the functions of the impaired gland, presumably the migrated foreign stem cells will differentiate into glandular cells to function as part of the salivary gland. Here we illustrate the steps involved in performing the cannulation procedure on the mouse sub mandibular salivary gland via the Wharton duct. For this experiment, the trip and blue solution is injected into the gland through the opening of the Wharton's duct with an ultra comfort 29 gauze needle and syringe.
Subsequently, the mouse is dissected to show that the infusions migrated into the gland successfully.Fully. Step one, one microliter per gram body weight of 0.5 milligrams per milliliter, atropine sulfate monohydrate is injected into each mouse subcutaneously to prevent the salivary secretions from disrupting the movement of the infusions. Step two, one microliter per gram body weight of a 100 milligram per milliliter, ketamine and 20 milligram per milliliter xylazine solution is injected into each mouse intraperitoneal to sedate the mouse.
Step three. After the sedation process, the mouse is placed on a platform. In the ventral position, the maxillary incisor is locked on a metal wire and the mandibular incisor is hooked on an elastic string in order to hold the mouth open.
Step four, using an ultra comfort 29 Gau needle and syringe filtered 0.4%Trian blue solution is infused into the submandibular salivary gland through the Wharton ducted, which is located right beneath the tongue. The needle is now pointing towards the opening of the Wharton duct, and now has entered the duct to deliver the infusions. Step five.
Immediately after the cannulation procedure, the salivary gland is surgically exposed in order to determine how much of the infusions have actually remained in the gland. After the infusions were injected, the mouse was dissected to surgically expose the post cannulated salivary gland. As you can see in this image, the dark coloration of the Wharton duct gives strong evidence that the trippin blue solution was infused successfully in the case of the salivary gland hypofunction.
The local application, namely the injection of drugs and cells through this cannulation procedure, may be perceived as a more direct and effective way in reviving the functions of the parotid, submandibular, and sublingual salivary glands. As evident in the representative results the infusions, which in this case is the Trian blue solution, was successfully deposited into the mouse submandibular salivary gland via the Wharton's duct. Hence, this cannulation technique can be used as a reliable model for the regenerative medicine of mouse salivary gland.
And as a future prospect, this procedure may be an expedient method for clinical gene transfer application. It would be a great pleasure for this visual article to be used as an effective tool for students and researchers who are interested in further studies in the regenerative medicine of the salivary gland.
