Gtx130274

Sections were undergone antigen retrieval with sodium citrate buffer and blocked by 3% goat serum (16210064, Gibco, USA) for 1 h. Then primary antibodies (Rabbit anti-CD31, 1:200, GTX130274, Gene Tex; Rabbit anti-Ki-67, 1:300, 12202 T, Cell Signaling Technology; Rabbit anti-CD163, 1:300, ab182422, Abcam) diluted in blocking buffer were used to incubate at 4 °C overnight. After washing with PBS, the Cell and Tissue Staining Kit (CTS005, Anti-Rabbit HRP-DAB System, R&D Systems) was used to detect the positive staining area. Images were taken by a microscope and analyzed by Image J software.

For histological analysis, tissues were fixed in formalin, embedded with paraffin, sliced into 5 μm sections, and stained using hematoxylin (3008-1, Muto Pure Chemicals, Tokyo, Japan) and eosin (32042, Muto Pure Chemicals, Tokyo, Japan) (H&E). Some tissue sections were deparaffinized and rehydrated prior to staining with CD31 antibody (GTX130274, Genetex, Hsinchu, Taiwan) using an automated immunohistochemistry system (BOND-MAX, Leica, Wetzlar, Germany). These were then visualized under a light microscope (IM-3, OPTIKA, Ponteranica, Italy). Wound parameters including wound edge distance, epithelial gap, granulation tissue area, re-epithelialization, epithelial tissue area, and adipose tissue gap were calculated from five individual mice.