Silica nanoparticles (SNPs) are produced on an industrial scale and are an addition to a growing number of commercial products. SNPs also have great potential for a variety of diagnostic and therapeutic applications in medicine. Contrary to the well-studied crystalline micron-sized silica, relatively little information exists on the toxicity of its amorphous and nano-size forms. Because nanoparticles possess novel properties, kinetics and unusual bioactivity, their potential biological effects may differ greatly from those of micron-size bulk materials. In this review, we summarize the physico-chemical properties of the different nano-sized silica materials that can affect their interaction with biological systems, with a specific emphasis on inhalation exposure. We discuss recentin vitroandin vivoinvestigations into the toxicity of nanosilica, both crystalline and amorphous. Most of thein vitrostudies of SNPs report results of cellular uptake, size- and dose-dependent cytotoxicity, increased reactive oxygen species levels and pro-inflammatory stimulation. Evidence from a limited number ofin vivostudies demonstrates largely reversible lung inflammation, granuloma formation and focal emphysema, with no progressive lung fibrosis. Clearly, more research with standardized materials is needed to enable comparison of experimental data for the different forms of nanosilicas and to establish which physico-chemical properties are responsible for the observed toxicity of SNPs.

Information on the toxicity of carbon nanotubes is still fragmentary but indicates that these particles can induce adverse effects. We previously demonstrated in rats that, when purified multi-wall carbon nanotubes (MWCNT) reach the lung, they are biopersistent and induce lung inflammation as well as fibrosis. The present study was designed to address the genotoxic potential of this material in the same species. In vivo, micronuclei (MN) were assessed in type II pneumocytes 3 days after a single intra-tracheal administration of MWCNT (0.5 or 2 mg). We also used the cytokinesis-block micronucleus assay in rat lung epithelial cells exposed in vitro to MWCNT (10, 25, 50 μg/ml). Finally, we applied a human pancentromeric fluorescent probe (fluorescent in situ hybridization assay) to differentiate clastogenic and/or aneugenic mechanisms in a human epithelial cell line (MCF-7). In vivo, we found a significant and dose-dependent increase in micronucleated pneumocytes after a single administration of MWCNT (∼a 2-fold increase at the highest dose). In vitro, we observed a significant increase of MN in epithelial cells after exposure to MWCNT (up to a 2-fold increase at the cytotoxic dose of 50 μg/ml). Finally, we found that MWCNT induced both centromere-positive and -negative MN in MCF-7 cells. Overall, this study provides the first evidence of the potential of MWCNT to induce clastogenic as well as aneugenic events.

Background: Increasing environmental and occupational exposures to nanoparticles (NPs) warrant deeper insight into the toxicological mechanisms induced by these materials. The present study was designed to characterize the cell death induced by carbon black (CB) and titanium dioxide (TiO₂) NPs in bronchial epithelial cells (16HBE14o- cell line and primary cells) and to investigate the implicated molecular pathways. Results: Detailed time course studies revealed that both CB (13 nm) and TiO₂(15 nm) NP exposed cells exhibit typical morphological (decreased cell size, membrane blebbing, peripheral chromatin condensation, apoptotic body formation) and biochemical (caspase activation and DNA fragmentation) features of apoptotic cell death. A decrease in mitochondrial membrane potential, activation of Bax and release of cytochrome c from mitochondria were only observed in case of CB NPs whereas lipid peroxidation, lysosomal membrane destabilization and cathepsin B release were observed during the apoptotic process induced by TiO₂ NPs. Furthermore, ROS production was observed after exposure to CB and TiO₂ but hydrogen peroxide (H₂O₂) production was only involved in apoptosis induction by CB NPs. Conclusions: Both CB and TiO₂ NPs induce apoptotic cell death in bronchial epithelial cells. CB NPs induce apoptosis by a ROS dependent mitochondrial pathway whereas TiO₂ NPs induce cell death through lysosomal membrane destabilization and lipid peroxidation. Although the final outcome is similar (apoptosis), the molecular pathways activated by NPs differ depending upon the chemical nature of the NPs.

Because of their small size and large specific surface area (SA), insoluble nanoparticles are almost not affected by the gravitational force and are generally formulated in stable suspensions or sols. This raises, however, a potential difficulty in in vitro assay systems in which cells adhering to the bottom of a culture vessel may not be exposed to the majority of nanoparticles in suspension. J. G. Teeguarden et al., 2007, Toxicol. Sci. 95, 300–312 have recently addressed this issue theoretically, emphasizing the need to characterize the effective dose (mass or number or SA dose of particles that affect the cells) which, according to their model based on sedimentation and gravitation forces, might only represent a very small fraction of the nominal dose. We hypothesized, in contrast, that because of convection forces that usually develop in sols, the majority of the particles may reach the target cells and exert their potential toxicity. To address this issue, we exposed three different cell lines (A549 epithelial cells, EAHY926 endothelial cells, and J774 monocyte-macrophages) to a monodisperse suspension of Stöber silica nanoparticles (SNP) in three different laboratories. Four different end points (lacticodehydrogenase [LDH] release, LDH cell content, tetrazolium salt (MTT), and crystal violet staining) were used to assess the cell response to nanoparticles. We found, in all cell lines and for all end points, that the cellular response was determined by the total mass/number/SA of particles as well as their concentration. Practically, for a given volume of dispersion, both parameters are of course intimately interdependent. We conclude that the nominal dose remains the most appropriate metric for in vitro toxicity testing of insoluble SNP dispersed in aqueous medium. This observation has important bearings on the experimental design and the interpretation of in vitro toxicological studies with nanoparticles.