Burn wound healing is an essential challenge in global healthcare systems, and there is enormous demand for advanced wound dressings to treat burn injuries. Bioactive electrospun mats based on polymers with antimicrobial performance are promising options to prevent wound infections and accelerate the wound healing process. In this study, a nanofibrous mat composed of polyvinyl alcohol/Alginate (ALG)/Arnebia Euchroma (AE) extract was fabricated by electrospinning technique. Then the prepared mat was characterized for morphology and chemical composition. The analysis demonstrated the presence of AE extract in nanofibrous mats with a uniform and unbranched appearance and a fiber diameter of around 100–200 nm. Also, the mechanical properties investigation shows promising results and in range of human skin. In addition, weight loss and water uptake abilities, water contact angle, and antibacterial activity were measured to verify mat's potential for wound healing. The in vivo study on burn wounds revealed excellent wound healing ability of the mats containing both ALG and AE extract with 92.5% wound closure after 21 days via highest collagen synthesis, which re-epithelization and remodeling the burned skin. Consequently, PVA/ALG/AE nanofibrous mats can be used as a promising wound dressing treat burn injuries.