Silver nanoparticles (AgNPs) were green synthesized using ethanolic extract of fenugreek leaves and characterized using UV-Vis spectroscopy, Fourier transform infra-red spectroscopy (FTIR), X-ray diffraction (XRD), high resolution transmission electron microscopy HRTEM and energy dispersive X-ray analysis (EDX) techniques. The HRTEM results revealed the formation of highly stable, mono dispersed, spherical shaped AgNPs with the size ranging from 20 to 30nm. The presence of flavonoids and their interaction with the AgNPs were confirmed using FTIR. Antibacterial activities of the AgNPs were studied against pathogenic gram-positive Staphylococcus aureus (S. aureus) and gram negative Escherichia coli (E. coli) bacteria. The synthesized AgNPs displayed the enhancement of antibacterial activity against E. coli. The morphological changes in the bacterial cell membrane was observed using SEM analysis. Leakage of protein from the bacterial cells increased at every time intervals (2 and 4h). MTT assay was carried out for the AgNPs against human skin cell line (HaCaT). Interestingly, cytotoxicity of the synthesized AgNPs was less toxic to HaCaT cells as compared to bacteria cells, which suggests that the synthesized AgNPs by this method is eco-friendly in nature.