Comparison of the DNA damage response in BEAS-2B and A549 cells exposed to titanium dioxide nanoparticles.

For some decades production of titanium dioxide nanoparticle (TiO2-NP) has been increasing at a considerable rate; concerns as to the toxicity of these particles upon inhalation have been raised. Indeed, TiO2-NPs have been shown to induce significant genotoxicity and to adversely affect both major DNA repair mechanisms: base excision repair (BER) and nucleotide excision repair (NER). The aims of the present study were to (i) compare the genotoxicity of TiO2-NPs and their impact on DNA repair processes on A549 alveolar carcinoma and BEAS-2B normal bronchial lung cell lines and (ii) delve deeper into the mechanisms leading to these effects. To achieve these goals, TiO2-NPs effects on cytotoxicity, genotoxicity, DNA repair activity and DNA repair gene expression were investigated in both cell lines upon exposure to 1-100 µg/mL of anatase/rutile, 21 nm TiO2-NPs. Our results show that TiO2-NPs induce comparable cytotoxic and genotoxic responses in BEAS-2B and A549 cells. Functional response to DNA damage is observed in both cell lines and consists of an overall downregulation in DNA repair processes. When evaluating the relative importance of the two DNA repair pathways, we observed a lower impact on BER compared with NER activities, suggesting that repair of oxidatively generated DNA damage is still triggered in these cells. This response becomes measureable at 4 h of exposure in BEAS-2B but only after 48 h of exposure in A549 cells. The delayed response in A549 cells is due to an initial overall and intense downregulation of the genes encoding DNA repair proteins. This overall downregulation correlates with increased methylation of DNA repair gene promoters and downregulation of NRF2 and BRCA1, which may thus be considered as upstream regulators. These results strengthen the evidence that TiO2-NP induces indirect genotoxicity in lung cells, via modulation of DNA repair processes, and shed some light on the mechanisms behind this effect.

Exposure-dependent Ag+ release from silver nanoparticles and its complexation in AgS2 sites in primary murine macrophages.

Silver nanoparticle (AgNP) toxicity is related to their dissolution in biological environments and to the binding of the released Ag(+) ions in cellulo; the chemical environment of recombined Ag(+) ions is responsible for their toxicological outcome, moreover it is indicative of the cellular response to AgNP exposure, and can therefore shed light on the mechanisms governing AgNP toxicity. This study probes the chemistry of Ag species in primary murine macrophages exposed to AgNPs by making use of X-ray Absorption Fine Structure spectroscopy under cryogenic conditions: the linear combination analysis of the near-edge region of the spectra provides the fraction of Ag(+) ions released from the AgNPs under a given exposure condition and highlights their complexation with thiolate groups; the ab initio modelling of the extended spectra allows measuring the Ag-S bond length in cellulo. Dissolution rates depend on the exposure scenario, chronicity leading to higher Ag(+) release than acute exposure; Ag-S bond lengths are 2.41 ± 0.03 Å and 2.38 ± 0.01 Å in acute and chronic exposure respectively, compatible with digonal AgS2 coordination. Glutathione is identified as the most likely putative ligand for Ag(+). The proposed method offers a scope for the investigation of metallic nanoparticle dissolution and recombination in cellular models.

Impact of anatase and rutile titanium dioxide nanoparticles on uptake carriers and efflux pumps in Caco-2 gut epithelial cells.

TiO2 microparticles are widely used in food products, where they are added as a white food colouring agent. This food additive contains a significant amount of nanoscale particles; still the impact of TiO2 nanoparticles (TiO2-NPs) on gut cells is poorly documented. Our study aimed at evaluating the impact of rutile and anatase TiO2-NPs on the main functions of enterocytes, i.e. nutrient absorption driven by solute-liquid carriers (SLC transporters) and protection against other xenobiotics driven by efflux pumps from the ATP-binding cassette (ABC) family. We show that acute exposure of Caco-2 cells to both anatase (12 nm) and rutile (20 nm) TiO2-NPs induce early upregulation of a battery of efflux pumps and nutrient transporters. In addition they cause overproduction of reactive oxygen species and misbalance redox repair systems, without inducing cell mortality or DNA damage. Taken together, these data suggest that TiO2-NPs may increase the functionality of gut epithelial cells, particularly their property to form a protective barrier against exogenous toxicants and to absorb nutrients.

In vitro evaluation of SiC nanoparticles impact on A549 pulmonary cells: cyto-, genotoxicity and oxidative stress.

Silicon carbide (SiC) is considered a highly biocompatible material, consequently SiC nanoparticles (NPs) have been proposed for potential applications in diverse areas of technology. Since no toxicological data are available for these NPs, the aim of this study was to draw their global toxicological profile on A549 lung epithelial cells, using a battery of classical in vitro assays. Five SiC-NPs, with varying diameters and Si/C ratios were used, and we show that these SiC-NPs are internalized in cells where they cause a significant, though limited, cytotoxic effect. Cell redox status is deeply disturbed: SiC-NP exposure cause reactive oxygen species production, glutathione depletion and inactivation of some antioxidant enzymes: glutathione reductase, superoxide dismutase, but not catalase. Finally, the alkaline comet assay shows that SiC-NPs are genotoxic. Taken together, these data prove that SiC-NPs biocompatibility should be revisited.

In vitro investigation of oxide nanoparticle and carbon nanotube toxicity and intracellular accumulation in A549 human pneumocytes.

If released in the environment, nanomaterials might be inhaled by populations and cause damage to the deepest regions of the respiratory tract, i.e., the alveolar compartment. To model this situation, we studied the response of A549 human pneumocytes after exposure to aluminium oxide or titanium oxide nanoparticles, and to multi-walled carbon nanotubes. The influence of size, crystalline structure and chemical composition was investigated. After a detailed identification of nanomaterial physico-chemical characteristics, cells were exposed in vitro and viability and intracellular accumulation were assessed. In our conditions, carbon nanotubes were more toxic than metal oxide nanoparticles. Our results confirmed that both nanotubes and nanoparticles are able to rapidly enter into cells, and distribute in the cytoplasm and intracellular vesicles. Among nanoparticles, we demonstrate significant difference in biological response as a function of size, crystalline phase and chemical composition. Their toxicity was globally lower than nanotubes toxicity. Among nanotubes, the length did not influence cytotoxicity, neither the presence of metal catalyst impurities.

Continuous production of water dispersible carbon-iron nanocomposites by laser pyrolysis: application as MRI contrasts.

Carbon encapsulated iron/iron-oxide nanoparticles were obtained using laser pyrolysis method. The powders were processed to produce stable and biocompatible colloidal aqueous dispersions. The synthesis method consisted in the laser decomposition of an aerosol of ferrocene solution in toluene. This process generated, in a continuous way and in a single step, a nanocomposite formed by amorphous carbon nanoparticles of 50-100 nm size in which isolated iron based nanoparticles of 3-10 nm size are located. The effect of using different carriers and additives was explored in order to improve the efficiency of the process. The samples after purification by solid-liquid extraction with toluene, were oxidised in concentrated nitric acid solution of sodium chlorate, washed and finally ultrasonically dispersed in 1 mM tri-sodium citrate solutions. The dispersions obtained have hydrodynamic particle size less than 150 nm and are stable in the pH range of 2-11. Finally the shortening of the transversal relaxation time of water protons produced by the dispersed particles was studied in order to test the feasibility of these systems to be traced by magnetic resonance imaging techniques.

TiC nanocrystal formation from carburization of laser-grown Ti/O/C nanopowders for nanostructured ceramics.

Refractory carbide ceramics (TiC and ZrC) raise interest as promising materials for high-temperature applications such as structural materials for the future generation of nuclear reactors. In this context, nanostructured ceramics are expected to exhibit improved thermomechanical properties as well as better behavior under irradiation when compared to conventional materials. It is therefore necessary to synthesize carbide nanocrystals of such materials to elaborate the ceramics. We report here the formation study of TiC nanocrystals through the direct carburization of Ti/O/C nanopowders grown by laser pyrolysis. A spray of titanium tetraisopropoxide was laser pyrolyzed with ethylene as the sensitizer, leading to Ti/O/C nanopowders with various C contents controlled by the synthesis conditions. Annealing treatments performed on these nanopowders under an inert atmosphere without any C addition enabled the formation of TiC grains through the carburization of the oxide phase by free C incorporated during the synthesis. The powders were characterized by X-ray diffraction, scanning electron microscopy, and transmission electron microscopy. The final TiC grain size was about 80 nm, and the grains were monocrystalline. The influence of the free C content on the grain growth during the annealing step, together with its effects on the densification of the ceramics after sintering by high-pressure flash sintering, was examined. A 93% densification was finally achieved.

Optical properties of synthetic carbon nanoparticles as model of cosmic dust.

Carbon nanoparticles synthesised by laser pyrolysis of small hydrocarbons are deposited at low energy on a silicon substrate. Infrared spectroscopy of the as-formed films are studied as a function of the synthesis parameters and post-treatments, such as annealing and heavy ion irradiation. Correlation between infrared spectroscopy and multiscale organisation of the samples is made through transmission electron microscopy, including image analysis. Changes in infrared spectra are analysed in terms of the carbon network building. The relevance of the results to model the structure and spectroscopy of carbon dust in the carbon-rich circumstellar media is discussed.