Background: Vernonia amygdalina is generally known as sour leaf or bitter leaf which is one among the medically significant plants in African and Mediterranean regions. Hence, this study aids the biosynthesis of copper nanoparticles using V.amygdalina for the first time and further it aims to investigate the cytotoxicity, antioxidant and anti-inflammatory properties of the V.amygdalina mediated copper nanoparticles. Methods: Cytotoxic effect of Copper sulphate nanoparticles reinforced with Vernonia amygdalina extract were assessed using Brine Shrimp Assay at 5 µL, 10 µL, 20 µL, 40µL and 80µL, anti-inflammatory activity using Bovine Serum Albumin (BSA) and antioxidant activity using DPPH Assay at 5 µL, 10 µL, 20 µL, 30 µL, 50 µL. Results: As the concentration increased, the cytotoxicity of the nanoparticles increased. It was found that the values for anti-inflammatory properties of nanoparticles was higher than the standard values at 30µL, 40µL, 50 µL concentrations. Percentage of inhibition was highest at 40 µL (85.7%) and 50 µL (92.6%). The values for antioxidant properties of nanoparticles were found to be higher than the standard values at 10 µL and 20 µL and almost equal in 30 µL concentrations. Conclusion: Vernonia amygdalina mediated copper nanoparticles have very less cytotoxic effects. It also has a potential to serve as a good anti-inflammatory and antioxidant agent.