| Summary: | This study explored the in vivo wound healing potential of <i>Vitis vinifera</i> seed extract using an excision wound model with focus on wound healing molecular targets including TGFBR1, VEGF, TNF-<i>α</i>, and IL-1<i>β</i>. The wound healing results revealed that <i>V. vinifera</i> seed extract enhanced wound closure rates (<i>p</i> < 0.001), elevated TGF-<i>β</i> and VEGF levels, and significantly downregulated TNF-<i>α</i> and IL-1<i>β</i> levels in comparison to the Mebo<sup>®</sup>-treated group. The phenotypical results were supported by biochemical and histopathological findings. Phytochemical investigation yielded a total of 36 compounds including twenty-seven compounds (<b>1</b>–<b>27</b>) identified from seed oil using GC-MS analysis, along with nine isolated compounds. Among the isolated compounds, one new benzofuran dimer (<b>28</b>) along with eight known ones (<b>29</b>–<b>36</b>) were identified. The structure of new compound was elucidated utilizing 1D/2D NMR, with HRESIMS analyses. Moreover, molecular docking experiments were performed to elucidate the molecular targets (TNF-<i>α</i>, TGFBR1, and IL-1<i>β</i>) of the observed wound healing activity. Additionally, the in vitro antioxidant activity of <i>V. vinifera</i> seed extract along with two isolated compounds (ursolic acid <b>34</b>, and <i>β</i>-sitosterol-3-<i>O</i>-glucopyranoside <b>36</b>) were explored. Our study highlights the potential of <i>V. vinifera</i> seed extract in wound repair uncovering the most probable mechanisms of action using in silico analysis.
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