Estefanía Conde

Species-specific toxicity of copper nanoparticles among mammalian and piscine cell lines.

Abstract The four copper nanoparticles (CuNPs) with the size of 25, 50, 78 and 100 nm and one type of micron-sized particles (MPs) (∼500 nm) were exposed to two mammalian (H4IIE and HepG2) and two piscine (PLHC-1 and RTH-149) cell lines to test the species-specific toxicities of CuNPs. The results showed that the morphologies, ion release and size of the particles all played an important role when investigating the toxicity. Furthermore, the authors found that the particle forms of CuNPs in suspensions highly contribute to the toxicity in all exposed cell lines whereas copper ions (Cu2+) only caused significant responses in mammalian cell lines, indicating the species-specific toxicity of CuNPs. This study revealed that the morphologies, ion release rate of NPs as well as the species-specific vulnerabilities of cells should all be considered when explaining and extrapolating toxicity test results among particles and among species.

Comparative cytotoxicity induced by bulk and nanoparticulated ZnO in the fish and human hepatoma cell lines PLHC-1 and Hep G2

Abstract The increasing presence of ZnO nanoparticles (NPs) in consumer products may be having a dramatic impact in aquatic environments. The evaluation of ZnO NP toxicity represents a great challenge. This study aimed at evaluating the cytotoxic effect of micro- and nanosized ZnO in a fish and a mammalian hepatoma cell line. A detailed characterisation of the particles in exposure media showed that ZnO NPs formed large aggregates. ZnO cytotoxicity was evaluated with a battery of in vitro assays including LUCS, a new approach based on DNA alteration measurements. In fish cells, ZnO NP aggregates contributed substantially to the cytotoxic effects whereas toxicity in the human cells appeared to be mainly produced by the dissolved fraction. ROS production did not contribute to the observed cytotoxicity. This work also showed that measuring concentrations of NPs is essential to understand the mechanisms underlying their toxicity.

Effects of cerium oxide nanoparticles to fish and mammalian cell lines: An assessment of cytotoxicity and methodology

Two cerium oxide nanoparticles (CeO(2) NPs) and one micro-sized CeO(2) particle were thoroughly characterized in their pristine form, in water and in cell culture medium. The particles were tested for cytotoxicity to the H4IIE rat hepatoma cell line or the RTG-2 rainbow trout gonadal cell line by means of four standard cytotoxicity assays. Nominal concentrations were verified by inductively coupled plasma mass spectrometry (ICP-MS) and methods were assessed for their suitability to detect reliably adverse effects due to particle exposure. All three particles showed aggregation in water and media. In the H4IIE cell line, the MTT cytotoxicity test revealed that negative effects could be observed for the CeO(2) NPs after 24h and for all particles after 72h of exposure, making the effects size, concentration and time dependent. No negative effect for the concentrations tested was detected for the remaining three assays and the RTG-2 cell line, making the MTT assay and the H4IIE cell line an appropriate system to assess adverse effects of CeO(2) NPs. A verification of the nominal concentration through ICP-MS revealed that there was a discrepancy between nominal and measured concentration depending on concentration and particle tested. Interferences of particles with assays were found to be present and need to be taken into consideration.

The potentiation effect makes the difference: non-toxic concentrations of ZnO nanoparticles enhance Cu nanoparticle toxicity in vitro.

Here we examined whether the addition of a non-toxic concentration (6.25 μg/mL) of zinc oxide nanoparticles (ZnONPs: 19, 35 and 57 nm, respectively) modulates the cytotoxicity of copper nanoparticles (CuNPs, 63 nm in size) in the human hepatoma cell line HepG2. The cytotoxic effect of CuNPs on HepG2 cells was markedly enhanced by the ZnONPs, the largest ZnONPs causing the highest increase in toxicity. However, CuNPs cytotoxicity was not affected by co-incubation with medium containing only zinc ions, indicating the increase in toxicity might be attributed to the particle form of ZnONPs. Transmission electron microscopy (TEM) revealed the presence of CuNPs and ZnONPs inside the cells co-exposed to both types of NP and outflow of cytoplasm through the damaged cell membrane. Inductively coupled plasma mass spectrometry (ICP-MS) determined an increase in the concentration of zinc and a decrease in that of copper in co-exposed cells. On the basis of these results, we propose that accumulation of large numbers of ZnONPs in the cells alters cellular membranes and the cytotoxicity of CuNPs is increased.

Tissue distribution of zinc and subtle oxidative stress effects after dietary administration of ZnO nanoparticles to rainbow trout.

The increasing use of ZnO nanoparticles (ZnO NPs) in different fields has raised concerns about the possible environmental risks associated with these NPs entering aquatic systems. In this study, using a dietary exposure route, we have analysed the tissue distribution and depuration pattern of Zn as well as any associated redox balance disturbances in rainbow trout (Oncorhynchus mykiss) following exposure to ZnO NPs (20-30nm). Fish were fed a diet spiked with ZnO NPs prepared from a dispersion in sunflower oil at doses of 300 or 1000mg ZnO NPs/kg feed for 10days. This uptake phase was followed by a 28days depuration phase in which fish from all groups received untreated feed. While no overt signs of toxicity were observed and no important effects in fish growth (weight and length) or in the hepatosomatic index among groups were recorded, we observed high levels of Zn bioaccumulation in the gills and intestine of exposed fish following exposure to both dose levels. Zn levels were not eliminated during the depuration phase and we have evidenced oxidative stress responses in gills associated with such long term ZnO NPs bioaccumulation and lack of elimination. Furthermore, exposures to higher doses of ZnO NPs (1000mg/kg feed) resulted in Zn distribution to the liver of fish following 10days of exposure. Fish from this exposure group experienced biochemical disturbances associated with oxidative stress in the liver and ethoxy-resorufin-O-deethylase (EROD) activity which may point to the ability of ZnO NPs or its ions to interfere with cytochrome P450 metabolic processes.

Mechanisms underlying the enhancement of toxicity caused by the coincubation of zinc oxide and copper nanoparticles in a fish hepatoma cell line

Ecosystems are exposed to a wide variety of individual substances, including at the nano-scale; and the potential adverse effects of their interactions are an increasing concern. The purpose of the present study was to determine whether zinc oxide nanoparticles (ZnONPs) at a no-observed-effect concentration modulate the cytotoxicity of copper nanoparticles (CuNPs) in the fish hepatoma cell line PLHC-1 after 48 h of exposure and the contribution of the released ions to these effects. Cells were exposed to 50-nm CuNPs (0.39-25.0 µg/mL), alone or in combination with ZnONPs (25 nm or 100 nm), at 6.25 µg/mL. Cells were exposed to suspensions of NPs or to their supernatants, as well as to their combinations. The effects on cell viability were assessed through cytotoxicity assays. Changes in cell morphology and metal internalization were also evaluated. The cytotoxicity exerted by CuNPs was enhanced in the presence of nontoxic concentrations of ZnONPs. On the contrary, Zn ions protected the cell line from the CuNP toxicity, this effect being related to an increase in the intracellular levels of Zn. This increase of metal was not observed in cells exposed to both ZnONPs and CuNPs, even when they were visualized inside the cell. The results indicated that the internalization of ZnONPs, but not the Zn ions, was responsible for the enhanced toxicity of the CuNPs. Environ Toxicol Chem 2016;35:2562-2570.

Quantification of the uranium concentration in human urine by inductively coupled plasma-sector field mass spectrometry (ICP-SFMS)

In this study, the validation of a method for analyzing the uranium (U) concentration in human urine samples by inductively coupled plasma-sector field mass spectrometry (ICP-SFMS) was conducted. PROCORAD (the Association for the Promotion of Quality Control in Radiotoxicological Analysis) provided two urine samples spiked with unknown contents of U (Sample A = 33.6 ± 1.0 µg/L and Sample B = 3.3 ± 0.1 µg/L) and one unspiked sample as a blank. The analyses were directly performed on the diluted urine samples (dilution factor = 1:20) in 5% v/v HNO3. The results obtained by ICP-SFMS corresponded well with the reference values, and the limits of detection were 235U = 0.049 × 10-3 µg/L and 238U = 7.37 × 10-3 µg/L. The ICP-SFMS technique has been shown to be successful in the analysis of the U concentration in human urine samples and for the quantification of isotopic ratios.