Nile tilapia (Oreochromis niloticus L.) ranks among Indonesia’s most heavily farmed freshwater fish, and the scales discarded during its processing are a rich, underused source of type I collagen, a structural protein of the extracellular matrix that supports skin repair and could serve as a natural ingredient in wound-care ointments. This study evaluated the physical properties of an ointment formulated with tilapia scale collagen extract and examined its effect on incision wound healing in mice (Mus musculus L.). Six groups were compared: K- (untreated), K+1 (bioplacenton), K+2 (ointment base only), and three collagen-ointment concentrations, P1 (5%), P2 (10%), and P3 (15%). Collagen was isolated by the acid-soluble collagen (ASC) method and characterized by Fourier-transform infrared (FTIR) spectroscopy; wound closure, tissue protein content (Lowry method), and hydroxyproline content were assessed over 14 days. All three ointment formulations met the general pharmaceutical standards for topical preparations. Wounds in every group closed gradually, reaching a mean closure of 87.4-98.2% by day 14 without a statistically significant difference among treatments, whereas protein and hydroxyproline assays showed that the 15% collagen ointment produced the lowest tissue protein (397.0 ± 8.55 μg/mL) and hydroxyproline (82.05 ± 4.45 μg/mL) levels, consistent with an accelerated transition into the tissue remodeling phase.