Abstract Amorphous silica nanoparticles (SiO2-NPs) have been studied for their toxic and genotoxic potential. Although contradictory data have been reported and the possible modes of action are not fully elucidated, aneugenic events have been reported, indicating the microtubule (MT) network as a potential target. To investigate this, we examined the effects of 59 nm (10 µg/ml) and 174 nm (7.5 µg/ml) SiO2-NPs on MTs in mitotic and interphase A549 human lung carcinoma cells. No gross morphological changes of the mitotic spindle or induction of multipolar spindles were observed upon SiO2-NPs treatment. The influence of SiO2-NPs on the interphase MTs network dynamics was investigated by in situ depolymerisation/repolymerisation experiments. Results showed a clear increase in MT dynamics after SiO2-NP treatment. Consistent with this, reduced levels of MT acetylation were observed. In addition, live cell microscopy demonstrated that SiO2-NP treatment reduced A549 cell motility. The SiO2-NP doses and conditions (serum-free) used in this study did not induce significant cell toxicity or MN frequencies. Therefore, the effects on MT dynamics, MT acetylation and migration observed, are direct effects of the SiO2-NPs and not a consequence of NP overload or toxic or genotoxic effects.