Development of environmentally friendly and natural-based products with healing properties has become a popular trend for human health. Here, for the first time as far as known, we reported an efficient antibacterial composite nanofiber which was functionalized with the herbal extract of Satureja hortensis (SH) via electrospinning technique for wound care dressing applications. The plant which was used for this purpose is cultivated on a commercial scale in our country, Turkey, and it is relatively inexpensive and widely used. Investigation of the herbal extract demonstrated 227% higher antibacterial activity against Staphylococcus aureus and 141% higher for Pseudomonas aeruginosa when compared with a commonly used strong antibiotic of amoxicillin. Thermoplastic polyurethane (TPU)-based composite nanofibers including SH with a diameter of about 734 nm for the wound dressings also showed efficient antibacterial activity against these two pathogens which can lead significant health problems and deaths. FTIR characterization confirmed that there is no chemical change of SH when it was incorporated to the composite polymeric nanofibers by the electrospinning technique. We strongly believe that the obtained results suggest a potential natural-based eco-friendly approach for wound care dressing in contrast with synthetic and metal based-antibacterial agents, which have many negative effects on human body and the environment.