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Background: Silver nanoparticles synthesized by the bio-green method have been applied to various biomedical applications. These procedures are simple, eco-friendly and serve as an alternative to complex chemical methods for the preparation of nanomaterials. Objective: In the present study, phytosynthesis of silver nanoparticles, to examine their antioxidant potential, toxic effects towards bacterial-, fungal-strains, brine shrimp nauplii and cancer cells was focused. Methods: Methanolic extract of Euphorbia wallichii roots was used for the synthesis of silver nanoparticles. The synthesis was monitored and confirmed by UV-visible spectroscopy, Fourier Transform Infra-Red (FTIR) spectrometric analysis, Field Emission Scanning Electron Microscope (FESEM), Energy Dispersive X-ray (EDX) and X-Ray Powder Diffraction (XRD). Results: The synthesized particles were average 63±8 nm in size. Involvement of phenolic (46.7±2.4 μg GAE/mg) and flavonoid (11.7±1.2 μg QE/mg) compounds as capping agents was also measured. Nanoparticles showed antioxidant properties in terms of free radical scavenging potential (59.63±1.0 %), reducing power (44.52±1.34 μg AAE/mg) and total antioxidant capacity (60.48±2.2 μg AAE/mg). The nanoparticles showed potent cytotoxic effects against brine shrimp nauplii (LD50 66.83 μg/ml), proliferation and cell death of HeLa cells as determined by MTT (LD50 0.3923 μg/ml) and TUNEL assays, respectively. Antimicrobial results revealed that silver nanoparticles were found to be more potent against pathogenic fungal (maximum active against A. fumigatus, MIC 15 μg/disc) and bacterial strains (maximum active against S. aureus, MIC 3.33 μg/disc) than the E. wallichii extract alone. Conclusion: These results support the advantages of using an eco-friendly and cost-effective method for synthesis of nanoparticles with antioxidant, cytotoxic and antimicrobial potential.