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The aim of this study was to design a dual-layer wound dressing as a new fibrous biomaterial based on the valorization of animal-derived proteins. The first layer was fabricated by the deposition of poly(ethylene oxide) (PEO) loaded with keratin hydrolysate (KH) via a mono-electrospinning process onto a poly(lactic acid) (PLA) film, which was used as a support. The second layer consisted of encapsulating a bovine collagen hydrolysate (CH) into poly(vinyl pyrrolidone) (PVP) through a coaxial electrospinning process, which was added onto the previous layer. This assemblage was characterized by electronic microscopy for morphology and the controlled release of KH. In vitro biocompatibility was evaluated on the L929 (NCTC) murine fibroblasts using quantitative MTT assay and qualitative cell morphological examination after Giemsa staining. Additionally, in vivo biocompatibility methods were used to assess the impact of the biomaterial on white Swiss mice, including the evaluation of hematological, biochemical, and immunological profiles, as well as its impact on oxidative stress. The results revealed a nanofibrous structure for each layer, and the assembled product demonstrated antioxidant activity, controlled release of KH, a high degree of in vitro biocompatibility, negligible hematological and biochemical changes, and minimal impact of certain specific oxidative stress parameters compared to the use of patches with textile support.