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The remarkable antibacterial activity and potential biomedical applications of silver nanoparticles (AgNPs) synthesized through eco-friendly approaches have garnered significant attention in recent years. This research aimed to introduce a green synthesis method for AgNPs using Salvia tebesana (S. tebesana) Bunge extract and to investigate their properties, antibacterial activities, and cytotoxicity effects. The fabricated AgNPs were analyzed with various analyses, including UV–Vis, DLS, XRD, FT-IR, and TEM analysis. The broth microdilution assay was applied to measure the Minimum Inhibitory Concentrations (MIC) of chemical AgNPs, biosynthesized AgNPs, and S. tebesana Bunge leaves extract alone against selected standard bacteria strains. The biosynthesized AgNPs displayed a surface plasmon resonance peak at approximately 415 nm, and the AgNPs synthesized using S. tebesana Bunge extract had a spherical configuration with an average size in the range of 10–15 nm. Biosynthesized nanoparticles showed a noteworthy antibacterial activity compared to chemical nanoparticles, particularly against P. aeruginosa, with the highest antibacterial activity reported at a MIC value of 39.06 μg/mL. AgNPs synthesized using S. tebesana Bunge extract demonstrated a significant decrease in fibroblast cell viability, but only when the concentration reached 2 mg/mL. The findings demonstrate that S. tebesana Bunge leaves extract enhances the antibacterial properties of AgNPs, and also represents an appropriate and biocompatible option for the synthesis of these nanoparticles. Our research highlights the potential of employing bio-safe and eco-friendly AgNPs synthesized in the presence of S. tebesana Bunge extract, which possess remarkable antibacterial properties, for various biomedical applications.