Although the use of thermoplastic polyurethane (Tpu) nanofiber mats as wound dressings is of great interest due to their mechanical properties, they are hindered by their poor wettability and bioavailability. In this study, we aimed to improve the cellular affinity of Tpu nanofiber mats for skin disorders by incorporating extracted collagen (Col) from tendons and physically mixed with a layer of phytoceramides (Phyto) to produce TpuCol@X-Phyto mats in which the weight % of Phyto relatively to the weight of the solution was X = 0.5, 1, or 1.5 wt% via facile electrospinning approach. The collective observations strongly indicate the successful incorporation and retention of Phyto within the TpuCol architecture. An increase in the Phyto concentration decreased the water contact angle from 69.4° ± 3.47° to 57.9° ± 2.89°, demonstrating improvement in the hydrophilicity of Tpu and binary blend TpuCol nanofiber mats. The mechanical property of 1.0 wt% Phyto aligns with practical requirements owing to the presence of two hydroxyl groups and the amide linkage likely contributing to various hydrogen bonds, providing mechanical strength to the channel structure and a degree of rigidity essential for transmitting mechanical stress. The proliferation of human skin fibroblast (HSF) peaked significantly 100 % with TpuCol@X-Phyto mats coated for X =1.0 and 1.5 wt% of Phyto. Electrospun scaffolds with the highest Phyto content have shown the lowest degree of hemolysis, demonstrating the high level of compatibility between them and blood. The TpuCol@1.5Phyto mat also demonstrated higher efficacy in antibacterial and antioxidant activities, achieving a rate of DPPH radical scavenging of 83.3 % for this latter property. The most notable wound closure among all tested formulations was attributed to higher Phyto. Thus, the developed TpuCol@1.5Phyto nanofiber formula exhibited enhanced healing in an in vitro epidermal model.