Zoletil vet

For the in vivo wound healing model, db/db mice were anesthetized by intraperitoneal injection of a mixture of zolazepam (Zoletil vet, Virbac, Taipei, Taiwan) and xylazine (Rompun, Bayer, Germany; 4:1, v/v). Two identical, circular, full-thickness cutaneous wounds in a diameter of 13 mm were surgically created on each side of the dorsum. Each wound margin was splinted with an O-ring by sutures to prevent the wounds from contraction. In each mouse, the wound on the left side was applied with 100 μL PBS as controls, and the wound on the right side was treated with 5 × 10⁵ ASCs (dbHG or dbLG) suspended in 100 μL PBS. A transparent, semiocclusive adhesive dressing (Tegaderm; 3 M, St. Paul, Minnesota) was applied over the wounds for protection. The wounds were photographed regularly postoperatively, and the wound size was measured through planimetric methods using ImageJ. The db/db mice were euthanized by 50% CO₂ and harvested entire wound tissues, including some of the surrounding skin area, on the postoperative days 5 and 21 for further analysis (n = 3 for each time point in each group).