Nanomaterials had been used because of the properties they exert in such scale (<100 nm), and they have been used in a wide variety of products like paints, electronics, fabrics, and also personal care products. Recent manuscripts available in the literature demonstrate the potential benefits of nanotechnology with these products. However, when released in the environment or when in contact with the human body, it is hard to track their final destination and their influence over the living beings. So, since nanomaterials were considered an important technology, a concern about its risks also started. Due to the variety of sizes, physicochemical properties, and uses, many researchers are aiming to assess the possible toxicity of this class of particles. Because of that, the chapter objective is to gather which assay, performed in vivo and in vitro, is the most frequently used and recommended to measure nanomaterial toxicity. Also, it is important to know which is the most suitable test to evaluate the toxicity over the environment, through direct effect and after biodegradation, and also related to human health. This chapter presents a concise review about the accepted methods to assess nanomaterial toxicity and also discuss about the need for regulamentation.