The growing problem of fishery waste poses a significant environmental concern and represents a valuable untapped source of biomaterials that can be converted into useful resources. This research aims to achieve the sustainable valorization of Cynoglossus arel waste. We extracted and characterized gelatin and chondroitin sulfate from fish skin and rigorously analyzed them to assess the integrity of their functional groups and their colloidal stability. The findings further verify that the gelatin has appropriate structure characteristics, as supported by tests for moisture content and gelation. Meanwhile, chondroitin sulfate was shown to be sulfated using Toluidine Blue, Safranin O, and Methylene Blue precipitation tests. Following their confirmatory tests, the antioxidant activities of the biopolymers were examined by the DPPH radical scavenging assay. The gelatin and chondroitin sulfate exhibited moderate antioxidant activity, suggesting their potential to combat oxidative stress. Their potential for biomedical applications, especially in wound healing, was evaluated using cell migration assays with normal human dermal fibroblast cells. When applied at specific concentrations, the biopolymer extracts significantly improved cell migration and accelerated wound healing compared with the control group. These results illuminate the high potential for utilizing Cynoglossus arel fish waste to produce bioactive compounds with significant therapeutic uses. The research establishes the dual advantage of valorizing marine waste and producing low-cost, natural biomaterials for regenerative medicine, especially for dermal repair.