Abstract: Pig hoof nails represent an underutilized byproduct of pork processing. According to traditional Chinese medical theory, pig hoof nails are believed to contain compounds that may exhibit pharmacological properties such as promoting hemostasis, aiding coagulation, and reducing inflammation, which could potentially benefit the healing of skin wounds. In this study, different molecular masses of hoof nail peptides (HNPs) were extracted from porcine hoof nails. Wound healing assays were employed to identify HNPs that significantly promote fibroblast proliferation and collagen synthesis across three molecular weight ranges: less than 10 kDa (HNP Ⅰ), 3-10 kDa (HNP Ⅱ), and more than 10 kDa (HNP Ⅲ). We formulated HNP Ⅱ into a temperature-sensitive gel designed to conform to uneven wound surfaces. This supports the development and application of hoof nail peptide formulations. The animal operation was approved by the Laboratory Animal Ethics Committee of Southwest University (No: IACUC-20231218-01). All experiments were conducted in accordance with relevant guidelines and regulations.