The rich phytochemical profile of Perilla frutescens leaf extract (PLE) makes it a compelling bioreducing agent for the green synthesis of silver nanoparticles (AgNPs), offering diverse biomedical applications. In this study, the PLE-mediated phytosynthesis of AgNPs was confirmed by UV-vis spectra with maximum absorbance at 440 nm. The spherical PLE-AgNPs of <20 nm in size were further characterized by Dynamic light scanning, Transmission electron microscopy, and Scanning electron microscopy. Moreover, High-resolution transmission electron microscopy, Selected area electron diffraction and X-ray diffraction validated the crystalline structure of PLE-AgNPs. The energy-dispersive X-ray and X-ray photoelectron spectroscopy established the presence of carbon, oxygen, and nitrogen groups, along with silver, in the PLE-AgNPs. FTIR results confirmed the presence of phytochemicals as the capping agents for PLE-AgNPs. The Gas chromatograph-mass spectrometry analysis revealed the presence of terpenoids, furan derivatives, phenolic compounds, hydroxides, imidazole, aldehydes, etc., in PLE. In addition, the presence of phytochemicals was also confirmed in the as-synthesized PLE-AgNPs. The 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity, hydrogen peroxide scavenging activity, and ABTS radical scavenging activity affirmed the strong radical scavenging effect of PLE-AgNPs with an IC50 of 51.58 μg·mL−1. The wound-healing potential of PLE-AgNPs was confirmed by in vitro cell scratch assay in human keratinocyte HaCaT cells. These integrated findings emphasize the significant biomedical advantages and potential applications of PLE-AgNPs in clinical applications.