Here, we explored the medicinal uses of the novel biogenic silver nanoparticles of Eugenia uniflora L. (E. uniflora) seed extract as a cost effective, eco-friendly, reducing and stabilizing compounds. This study describes the synthesis of silver nanoparticles from Eugenia uniflora L. (E. uniflora) seed extract and their antioxidant, antibacterial and cytotoxic potential. Biosynthesis of AgNPs was monitored by UV-visible spectroscopy which revealed intense surface plasmon resonance bands at 447 nm and X-ray diffraction were employed to identify various functional groups and crystalline nature of AgNPs. Scanning electron microscopy studies demonstrated that synthesized particles were crystalline in nature with average size of 78-100nm. In vitro antioxidant effects were analyzed by butylated hydroxytoluene (BHT)) and 2,2-diphenyl-1-picrylhydrazyl (DPPH), which exhibited antioxidant activity there in the particles could scavenges the stable free radical DPPH of 75% to that o positive control BHT. The value of 50% inhibition concentration (IC50) of Standard BHT is 65.55 and E. uniflora is 38.63µg/ml. The antibacterial activity of green AgNPs displayed better zone of inhibition against selected human pathogens. The present study also investigated the toxicity effect of biogenic AgNPs against human prostate cancer cells (PC-3) and the inhibitory concentrations (IC50) were found to be 6.25μg/ml, respectively. It could be concluded that E. uniflora seed extract AgNPs can be used efficiently for potential antioxidant, antibacterial and cytotoxic potential AgNPs with potent biomedical applications.