Nanocellulose is possessing the unique physical/structural properties, low product cost, renewable, biodegradable, and biocompatible material. For these characteristics, nanocellulose is applicable the many industrial fields such as energy, medical, packing and so on. Nanocellulose from wood and other higher plants is typically isolated as cellulose nanofibers (CNFs) and cellulose nanocrystals (CNCs). Specially, CNFs have strong water absorption capacity and thickening effect, which enhances the viscosity and apply to cosmetic applications. However, CNFs raise the safety concern because they have the high aspect ratio and fibrous morphology. For these reason, this study carried to cytotoxicity, skin irritation test and eye irritation test to find the cosmetic application of CNFs. As a result of this study, CNFs significantly induced cytotoxicity to HaCaT cells (≥ 156 µg/ml) and HDF-α cells (≥ 313 µg/ml). But, CNFs did not induce the skin and eye irritation on 3D models. Taken together, this study suggested that appropriate concentration setting is needed to usage of CNFs as the cosmetic material.