Cytotoxic evaluation of a mixture of eight pollutants at environmental relevant concentrations / Evaluación de la citotoxicidad de una mezcla de ocho contaminantes a concentraciones de relevancia ambiental

The ubiquitous presence of pollutants and the accurate evaluation of their potential risks for environmental and human health is an area of major concern. We have simulated an in vitro scenario of long-term exposure to a mixture of eight pollutants at real environmental concentrations using mammalian Vero cells. Our results demonstrate that cellular proliferation rates were significantly altered, either by inhibition or stimulation, depending on the mixture composition and the exposure time. We encourage the urgency of reviewing safety levels for emerging contaminants accepted by regulatory agencies, considering that mixtures of pollutants represent a threat for environmental and human health (AU)

La presencia ubicua de contaminantes ambientales y la adecuada evaluación de su riesgo potencial para la salud humana y ambiental es un área de gran preocupación. En este trabajo se ha simulado un escenario in vitro de exposición a largo plazo de una mezcla de ocho contaminantes a concentraciones reales presentes en el medio ambiente, utilizando la línea celular de mamífero Vero. Nuestros resultados demuestran que se alteran significativamente las tasas de proliferación celular, ya sea por estimulación o inhibición, dependiendo de la composición de la mezcla y del tiempo de exposición. En vista de estos resultados, recalcamos la necesidad de revisar los niveles de seguridad aceptados por las agencias reguladoras para contaminantes emergentes, teniendo en cuenta que las mezclas de contaminantes representan una amenaza para la salud humana y medioambiental (AU)

Toxicological evaluation of three contaminants of emerging concern by use of the Allium cepa test.

Di(2-ethylhexyl)phthalate, triclosan and propylparaben are contaminants of emerging concern that have been subjected to extensive toxicological studies, but for which limited information is currently available concerning adverse effects on terrestrial plant systems. The Allium cepa test, which is considered one of the most efficient approaches to assess toxic effects of environmental chemicals, was selected to evaluate the potential risks of these ubiquitous pollutants. Our data demonstrate that all three compounds studied may in some way be considered toxic, but different effects were noted depending on the chemical and the end point analysed. Results derived from the analysis of macroscopic parameters used in testing for general toxicity, revealed that while di(2-ethylhexyl)phthalate had no apparent effects, the other two chemicals inhibited A. cepa root growth in a dose-dependent manner. On the other hand, although all three compounds caused alterations in the mitotic index of root-tip cells, propylparaben was the only one that did not show evidence of genotoxicity in assays for chromosome aberrations and micronuclei. The results of the present study clearly indicate that sensitive plant bioassays are useful and complementary tools to determine environmental impact of contaminants of emerging concern.

An integrated cellular model to evaluate cytotoxic effects in mammalian cell lines.

The ever growing anthropogenic pressure to the environment has lead in 2007 to the revision of the existing legislation and the approval of the new European law regarding the production and importation of chemicals, known as REACH. This new legal framework supports the development of alternative methods to animal experimentation encouraging the improvement and/or design of new methodological strategies for the toxicological evaluation of chemical compounds. Even though cytotoxicity studies are a reductionist approach to acute toxicity in vivo, they offer the best agreement between obtaining relevant information about the mechanism of toxic action and the use of alternative methods. Following this trend, this work presents an integrated cellular strategy in order to know the toxicity and mechanism of action of chemical compounds, using simple and reproducible in vitro systems. The experimental procedures are performed in two steps. The first one involves the systematic analysis of the main cellular targets using proliferation, viability and morphological probes. The second step relies upon the results obtained in the first step, including specific assays that focus on the mechanism of toxic action and the cellular response. The benefits of this strategy are exemplified with two real cases: pentachlorophenol and rotenone.

In vitro assessment of the cytotoxic and mutagenic potential of perfluorooctanoic acid.

Perfluorooctanoic acid (PFOA) is a perfluorinated compound ubiquitously detected in the environment, including wildlife and humans. Despite the available information, research on the cytotoxicity of PFOA in non-tumoral mammalian cells is relatively limited. In this work, two in vitro toxicity systems were employed to provide further insight into the cytotoxic and mutagenic potential of PFOA. The cytotoxicity of the chemical towards Vero cells was assessed using biochemical and morphological parameters, while mutagenicity was evaluated according to Ames test. High doses of PFOA cause oxidative stress in Vero cells, that was closely linked to cell cycle arrest at the G1 phase and induction of apoptosis. Our results corroborate previous findings in human tumoral cells and suggest that the mode of action of this perfluorinated compound is not a peculiarity among mammalian cell types. On the other hand, the compound was not mutagenic in the Ames test, using four strains of Salmonella typhimurium in the presence or absence of rat S9 metabolic activation system.