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The green synthesis of B. tinctoria zinc oxide nanoparticles (ZnO NPs) was rendered by using leaves and fruits extract. These extracts acted as a capping and reducing agent in stabilizing the formation of ZnO NPs. The obtained NPs were characterized by using absorption spectroscopy analysis (UV-vis) and Fourier-transform infrared spectroscopy (FT-IR) which showed a distinct peak at 274 nm, 467cm-1 and 456 cm-1. Further, the formation has been confirmed by powder X-ray diffraction (XRD) and the obtained XRD pattern fitted well with the JCPDS card, showing the pure crystalline nature of synthesized NPs Patten. The Scanning Electron Microscopy (SEM) analysis revealed hexagonal particle shape and Energy Dispersive X-ray (EDX) by confirming the strong signals of Zinc and oxygen. The results of Dynamic Light Scattering (DLS) showed that NPs obtained from both NPs of 244nm and 256nm size with the surface zeta potentials of -15.0mV and -18.9mV. Antibacterial and anti-biofilm efficacy of synthesized nanoparticles were evaluated against six human pathogenic bacteria, resulted that antibacterial/ anti-biofilm activity was increased upon increasing the concentration dose range. The obtained NPs exhibited significant antioxidants potential, which was made to study the anticancer activity against Raw 264.7 and Caco-2 cancer cell lines, which showed absolvable activity.