Recently, nontoxic origin-mediated synthesis of copper oxide nanoparticles acquires further recognition because of the key role of bioapplications. The plant Cissus quadrangularis is one most prominent herbs used in the treatment of diabetes, asthma, tissue regeneration, etc. In this study, we tested the process of copper oxide nanoparticle synthesis and their role in many functions from Cissus quadrangularis. The synthesis of copper oxide nanoparticles uses plant extract and characterization by X-ray diffraction, thermogravimetric analysis (TGA), Fourier-transform infrared spectroscopy (FT-IR), atomic force microscope (AFM), and scanning electron microscope (SEM). The synthesized nanoparticles were analyzed for their biomedical applications such as antibacterial, antifungal, antioxidant, antidiabetic, and anti-inflammatory activity and antiproteinase action. The results show that the C. quadrangularis plant-mediated nanoparticles may be used in many biomedical applications related to arthritis, diabetes, and the production of various antimicrobial products in the future.