Wound Healing Model in Immunocompromised Mice

Luciferase/GFP-labeled GMSCs were implanted in wound healing model as described previously [22 (link)–24 (link)]. In brief, 5-week-old immunocompromised mice were individually anesthetized using an intraperitoneal injection of ketamine (75 mg/kg) and rinsed with an alcohol swab and sterilely prepped with betadine and draped. A sterile 8 mm diameter full-thickness wound was created on the dorsum of the nude. A donut-shaped splint with a 10 mm inner diameter and 20 mm outer diameter was fashioned from a 0.5 mm-thick silicone sheet (Grace Bio-Laboratories, Bend, OR). An immediate-bonding adhesive (Tegaderm, 3 M) was used to fix the splint to the skin followed by interrupted 5–0 nylon (Ethicon, Inc,Somerville, NJ) sutures to ensure position. Mastisol (Fernadale, MI) was applied to the perimeter of the wound to improve adherence of the occlusive dressing (Tegaderm, 3 M) placed to cover the wounds. The animals were placed in individual cages under a warming lamp and allowed to recover fully from anesthesia. The wound dressings in each group were changed every 3 days according the above methods. 15 mice were randomly divided into three groups: Group A, hydrogel/control GMSCs; Group B, hydrogel/center-BRONJ GMSCs; Group C, hydrogel/peri-BRONJ GMSCs, n = 5. Figure 4 showed the experimental design and schematic representation of wound healing model in nude mice.