eng Oriental Scientific Publishing Company Biosciences Biotechnology Research Asia 0973-1245 2025-12-30 22 4 1541 1563 10.13005/bbra/3459 56777 article Pooja Malik 1 Swapnil Sharma 1 Department of Pharmaceutics, School of Pharmaceutical Sciences, Faculty of Pharmacy, IFTM University, Moradabad, India. The current work aimed to develop and optimize a hydrogel comprising Morus alba (mulberry) leaf extract for effective topical administration with enhanced antibacterial properties. Morus alba is well-known for its antioxidant, anti-inflammatory, wound-healing, and antibacterial qualities, making it an attractive natural alternative for dermatological applications. Carbopol 934 served as the major gelling agent, with appropriate co-polymers, neutralisers, and humectants added to improve texture, stability, and release qualities. The ethanolic extract was shown to possess bioactive phytoconstituents after preliminary phytochemical screening. A 3² factorial design was used to optimise the concentration of polymer and penetration enhancer, resulting in nine hydrogel formulations (F1-F9). These were tested for key physicochemical properties such as pH, viscosity, spreadability, drug content, grittiness, and homogeneity. In vitro release tests using Franz diffusion cells revealed sustained release behaviour, whilst ex vivo permeation through rat skin using Franz diffusion cell of the optimised formulation (F9) showed a total drug release of 89.72% over 48 minutes. The antimicrobial activity of the optimised formulation against Escherichia coli, Staphylococcus aureus, and Candida albicans was determined using the agar well diffusion method. The hydrogel inhibited all pathogens tested significantly, with Staphylococcus aureus, E. coli, and C. albicans showing the highest efficiency. This investigation verified the formulation's ability to cure bacterial and fungal skin infections. The formulation was also discovered to be non-irritating and pH suitable for human skin. To summarise, the Morus alba extract-loaded hydrogel provides a stable, safe, and effective natural option for topical antimicrobial therapy by integrating current formulation technology with the therapeutic benefits of herbal medicine. https://www.biotech-asia.org/vol22no4/formulation-optimization-and-antimicrobial-evaluation-of-morus-alba-leaf-extract-based-hydrogel/ Antimicrobial activity; Factorial design; Hydrogel; Herbal formulation; Morus alba; Topical drug delivery