The increasing prevalence of chronic wounds, associated with aging, obesity, and diabetes, is compounded by bacterial resistance and adverse effects associated with commercial wound care products. Therefore, treatment modalities to accelerate the healing process are constantly being sought. This study aimed to investigate the phytochemical composition, antioxidant, and wound healing potential of the aqueous extract derived from the stems and leaves of Polygonum minus. The extract was subjected to phytochemical evaluation to assess the diversity of secondary metabolites. The total phenolic content (TPC) and total flavonoid content (TFC) were measured using the Folin-Ciocalteu colorimetric and aluminium chloride methods, respectively. The antioxidant activity was determined using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging and ferric-reducing antioxidant power (FRAP) assays. The wound healing effects were evaluated using proliferation and migration assays on human epidermal keratinocyte (HaCaT) cells. The phytochemical evaluation of the aqueous extract revealed the presence of flavonoids, terpenoids, alkaloids, saponins, tannins, steroids, and cardiac glycosides. Furthermore, the extract exhibited high TPC (137.74 ± 0.75 µg/mL GAE) and TFC (177.08 ± 3.16 µg/mL QE) values, as well as radical scavenging activity at 79.50% in the DPPH assay and a FRAP value of 1485.67 ± 0.05 µM/g Fe2+. At the lowest concentration of 7.81 μg/mL, the extract significantly stimulated cell proliferation and migration within 24 hours of treatment. The stimulation of cell migration was comparable with that of allantoin, which was used as a positive control. This study indicated that the P. minus aqueous extract contains a high concentration of phenolic compounds, which could contribute significantly to its antioxidant activity and promote the proliferation and migration of keratinocytes. The findings suggest that the extract may merit further investigation for potential applications in topical therapy related to wound healing. © 2018-2024, Rangsit University.