Zinc oxide nanoparticles (ZnO-NPs) are odorless, white, and insoluble in water and alcohol. There are various applications for ZnO due to its unique semiconducting, piezoelectric, and optical features. Biodegradability and low toxicity are important properties of ZnO nonmaterial. The OH groups on the ZnO surface allow different surface decorating molecules to easily functionalize it. Mineralized in muscle, brain, skin, bone, and other tissues, zinc is a necessary mineral. It is a major component of most enzyme systems and is involved in protein and macromolecule production, neurogenesis, and hematopoiesis, among other functions. Due to its small particle size, Nano-ZnO is easily absorbed by the body. So-called nano-ZnO is commonly utilized in food additives. Chemical surface properties, particle shape, size distribution, and particle reactivity in solution all influence the biological activity of nanoparticles. Determining the morphology, size, and functioning of nanoparticles with regulated structures is critical for biological applications. In this review article, biological and non-conventional synthesis and applications of ZnO-NPs are highlighted in terms of their biological activity. ZnO-NPs have shown therapeutic activity against cancer, diabetes, microbial infection, inflammation, and potential antioxidant activity, which can be used as drug carriers, imaging tools, and biosensors.