Nanotechnology is an emerging branch of science, which has potential to solve many problems in different fields. The union of nanotechnology with other fields of sciences including physics, chemistry, and biology has brought the concept of synthesis of nanoparticles from their respective metals. Till date, many types of nanoparticles have been synthesized and being used in different fields for various applications. Moreover, copper nanoparticles attract biologists because of their significant and broad-spectrum bioactivity. Due to the large surface area to volume ratio, copper nanoparticles have been used as potential antimicrobial agent in many biomedical applications. But the excess use of any metal nanoparticles increase the chance of toxicity to humans, other living beings, and environment. In this article, we have critically reviewed the bioactivities and cytotoxicity of copper nanoparticles. We have also focused on possible mechanism involved in its interaction with microbes.