Green synthesis is considered to be one of the most suitable method because it enhances the therapeutic effects of palladium nanoparticles (Pd NPs). In this study, various biological activities such as antimicrobial, anticancer, antioxidant, and DNA cleavage activities of Urtica-mediated green synthesizing Pd NPs were investigated. The synthesized Pd NPs were characterized by using UV-vis, XPS, FT-IR, TEM, and XRD analyses. As a result of the TEM analysis of Pd NPs, the mean particle size was found to be 7.44 +/- 1.94 nm, and this result was supported by XRD analysis. The maximum DPPH scavenging activity was determined as 79.6% at 500 mg/L. The newly green synthesized Pd NPs exhibited high antimicrobial activity to gram-negative bacteria than gram-positive bacteria. Urtica-mediated green synthesized Pd NPs also showed double strain DNA cleavage activity. For the cytotoxic effects of Pd NPs, the MDA-MB-231 breast cancer cell line, HT-29 colon cancer cell line, Mia Paca-2 human pancreatic cancer cell line, and healthy cell line L929-Murine fibroblast cell line were used. IC50 values of Pd NPs against MDA-MB-231, HT-29, and MIA PaCa-2 cancer cell lines were calculated as 31.175, 20.383, and 29.335 mu g/ml, respectively. No significant cytotoxic effect was observed in the healthy lines L929.