Uptake of TiO₂ Nanoparticles into <i>C. elegans</i> Neurons Negatively Affects Axonal Growth and Worm Locomotion Behavior

Abstract

We employ model organism Caenorhabditis elegans to effectively study the toxicology of anatase and rutile phase titanium dioxide (TiO₂) nanoparticles (NPs). The experimental results show that nematode C. elegans can take up fluorescein isothiocyanate-labeled TiO₂ NPs and that both anatase and rutile TiO₂ NPs can be detected in the cytoplasm of cultured primary neurons imaged by transmission electron microscopy. After TiO₂ NP exposure, these neurons also grow shorter axons, which may be related to the detected impeded worm locomotion behavior. Furthermore, anatase TiO₂ NPs did not affect the worm's body length; however, we determined that a concentration of 500 μg/mL of anatase TiO₂ NPs reduced the worm population by 50% within 72 h. Notably, rutile TiO₂ NPs negatively affect both the body size and worm population. Worms unable to enter the L4 larval stage explain a severe reduction in the worm population at TiO₂ NPs LC₅₀/3d. To obtain a better understanding of the cellular mechanisms involved in TiO₂ NP intoxication, DNA microarray assays were employed to determine changes in gene expression in the presence or absence of TiO₂ NP exposure. Our data reveal that three genes (with significant changes in expression levels) were related to metal binding or metal detoxification (mtl-2, C45B2.2, and nhr-247), six genes were involved in fertility and reproduction (mtl-2, F26F2.3, ZK970.7, clec-70, K08C9.7, and C38C3.7), four genes were involved in worm growth and body morphogenesis (mtl-2, F26F2.3, C38C3.7, and nhr-247), and five genes were involved in neuronal function (C41G6.13, C45B2.2, srr-6, K08C9.7, and C38C3.7).