Novel nanoparticles (NPs) such as zinc oxide (ZnO) NPs are widely produced and applied in our daily lives at a rapid pace. Thus, the toxicity of ZnO NPs should be monitored as an important standard for environmental risk assessment. Here we assessed the in vitro cytotoxicity of ZnO NPs on human pulmonary adenocarcinoma cells LTEP-a-2 by tetrazolium salt colorimetric assay of cell proliferation in the presence or absence of ZnO NPs. ZnO NPs-induced morphological changes in LTEP-a-2 cells were examined by light and scanning electron microscopy. The mechanism by which ZnO NPs impose the cytotoxic effect was investigated by a combination of active oxygen test, lactose dehydrogenase-release assay, and apoptosis detection. Results showed that ZnO NPs significantly inhibited the proliferation and induced evident morphological changes (cell shrinkage and chromosome condensation) in LTEP-a-2 cells. Additionally, ZnO NPs increased the level of intracellular reactive oxygen species and induced the formation of apoptotic vesicles as well as the lysis of cell nuclei. Zn2+ ions released from ZnO NPs into aqueous solution are important components that exert cytotoxic effects on LTEP-a-2 cells. This study provides new insights to the cytotoxicity of ZnO NPs against human health. (C) 2014 The Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.