Propolis is a natural substance produced by bees from tree resins, and it has been widely studied for its potential health benefits. One area of interest is its effect on wound healing, as it has been shown to have antimicrobial, anti-inflammatory, and antioxidant properties. This study investigated propolis's toxicity and wound-healing properties and optimized the fabrication method for a potential wound-healing patch. The Prostoblue assay was used to evaluate propolis toxicity on Detroit 551 cells, and concentrations ranging from 0 to 12.5 µM were selected for the subsequent wound healing assay. The results showed that propolis at 2.5 µM was the most effective for wound healing in Detroit 551 cells, demonstrating a wound recovery rate of 93.22%. In addition to investigating the wound-healing properties of propolis, we optimized the beeswax and cotton gauze fabrication method for a wound-healing patch. Using olive oil as a plasticizer for beeswax resulted in even distribution throughout the gauze, providing a potential material for a wound-healing patch with antimicrobial, anti-inflammatory, and moisturizing effects. The impact of this study is significant as it provides preliminary evidence for the potential use of propolis in wound healing and identifies a potential fabrication method for a wound-healing patch. Further studies are warranted to investigate the efficacy of this material as a wound-healing patch and to optimize its formulation for maximum effectiveness. This study contributes to the growing body of research on natural products for wound healing and may lead to the development of more effective and sustainable wound-healing treatments. The promising prospects of this research are vast, as the wound healing patches are not only a sustainable alternative to synthetic healing patches but also have the potential to reduce and battle antibiotic resistance and broaden access to healthcare for patients in resource-limited areas.