Approved Abstracts

Cytotoxicity and mutagenicity effect of polystyrene microplastics and flame retardants compounds



Author(s): Andreu-Sánchez O; Andreu-Moliner E;
Presenter: Oscar Andreu-Sánchez

Emerging contaminants (ECs) are known substances whose presence in the environment is not necessarily new, but have not been included in the priority lists due to the absence of regulation or harmonized information on the levels and effects detected in ecosystems. Among them the brominated flame retardants (BFRs) stand out due to their release from a variety of point sources mainly from urban and industrial origin. The Tetrabromobisphenol A (TBBPA) is considered the dominant BFR present in industrial and consumer products. Plastics, which exist in a variety of chemical compositions, forms and particle sizes, are relatively novel and emerging actors in terrestrial and aquatic habitats worldwide accounting for 50-80% of marine debris. These substances are synthetic organic polymer particles derived from petroleum sources, including polystyrene (PS).
The particles tested were 3 polystyrenes (PS) with different particle diameter (PS1: 0.03 m, PS2: 0.6m and PS3: 3m (Sigma-Aldrich). The range of concentrations was between 100 - 1 mg/L for each of the PS. The tests were carried out using the compounds individually. Regarding the TBBPA (CAS number 79-94-7) the nominal concentrations assayed were in the range of 100-1 mg/L. The cytotoxicity effect has been evaluated according the MTT assay (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide). The assay is based on the conversion of MTT into formazan crystals by living cells, which determines mitochondrial activity. The human lung A549 and the human intestinal CaCo2 cell lines were used in the study. For Mutagenicity effect evaluation, the Ames test with both TA100 and TA98 bacterial strains of Salmonella thyphymurium have been carried out. The tests have been carried out by means of the “fluctuation test” in accordance with the OECD TG 471 by using the Mutachromotest kit (EBPI Inc, Canada). The assays were performed in sterile 96-well plates, the exposure time was 6 days. After that the reading of reverted wells (yellow) and unreverted wells (purple) was made. The results for Ames test indicated that there is some mutagenicity activity in the TA100 strain at concentrations of 1 and 10 ppm only for the PS1. The mutagenic activity for the TA98 strain is not clear at the range of indicated concentrations. It seems there is an interference between the PS and the culturing media that must be solved.
The results for MTT assays showed that only the PS1 had cytotoxicity effect on the A549 cell line. No effects on the A549 or CaCo2 cells were found for the rest of studied PS.

Acknowledgement: This study was supported by the Ministerio de Ciencia e Innovación. Project number: RTI2018-096046-B-C21-INTERTOX


Keywords: flame retardant; Citotox; Mutagenicity

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