Infectious diseases are becoming a worldwide threat due to antibiotic resistance. Therefore, it is necessary to prepare innovative antibiotics against bactericidal activities. The development of nanomedicine has been greatly aided by nanotechnology. A focal point of exploration involves the production of noble metal nanoparticles, owing to their diverse applications. The study's subjects are the synthesis, characterization, and assessment of silver nanoparticles (AgNPs) derived from Berberis lyceum Royle root bark (BLR) extract. The study revealed the potent antibacterial properties of BLR‐AgNPs, with heightened efficacy against S. pyogenes (16.7±0.3 mm) and comparatively lower activity against E. coli (1.1±0.1 mm). Minimum inhibitory concentration assessment indicated maximum activity at 10 μg/mL, whereas 2.5 μg/mL demonstrated the least inhibitory effect. Furthermore, the wound healing potential of BLR‐AgNPs was explored, demonstrating superior activity (0.19±0.02 cm) equated to BLR‐extract (0.46±0.02 cm) and untreated wounds (0.77±0.02 cm). These findings underscore the considerable antibacterial and wound‐healing capabilities of BLR‐AgNPs.