Approved Abstracts

Cytotoxic and Genotoxic Assessment of Zinc Oxide, Copper Oxide and Graphene Nanoparticles on the human lung cell line A549

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

The use of Zinc oxide (ZnO), Copper oxide (CuO) and graphene nanoparticles (NPs) gained a great interest in new emergent industrial applications such as inkjet printing or intelligent labelling. Nevertheless, this new uses posed an unevaluated risk for people involved in its life-cycle that must be evaluated. The aim of this work was to evaluate the cytotoxicity and genotoxicity of this nanoparticles on the A459 cell line. The HRTEM analysis showed that CuO nanoparticles were mainly spherical, with a primary diameter between 10 and 90 nm. ZnO nanoparticle were mainly composed by hexagonal nanocrystals of different sizes between 10-100 nm. In the case of Graphene, SEM analysis showed that nanoparticles were almost spherical and have a primary size range between 10 and 100 nm. The A549 cells are widely used as a type II pulmonary epithelial cell model for drug metabolism but recently it has been used in the evaluation of nanomaterial genotoxicity of NPs by inhalation route. Genotoxicity has been evaluated by Comet assay (CA), also called single cell gel electrophoresis (SCGE), detects ADN damage due to the ability of negatively charged DNA fragments to pass through agarose in response to an electrical flow. The cytotoxicity has been evaluated according the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay was selected to evaluate the NPs cytotoxicity effect. The assay is based on the conversion of MTT into formazan crystals by living cells, which determines mitochondrial activity. Respect de genotoxicity effect, the studied compounds showed a dose-dependent genotoxic effect on A549 cells. The highest effect was produced by CuO followed by ZnO and lowest effect was produced by graphene. The 24h-EC50 are, respectively: 1.3 mg/L, 2.1 mg/L and 3.1 mg/L. The statistical study (ANOVA) showed significant statistically differences (alpha=0.05) respect the untreated control The cytotoxicity assays indicated a higher toxicity for ZnO nanoparticles (24h-EC50= 21.5 mg/L), followed by CuO (24h-EC50= 35.9 mg/L) and finally Graphene (24h-EC50= 44.1 mg/L). The NOEC (No observed Effect Concentration) for each NP was calculated as 24h-EC10 . ZnO nanoparticles (24h-EC10= 3.9 mg/L), followed by CuO (24h-EC50= 7 mg/L) and Graphene (24h-EC50= 13.9 mg/L).

Keywords: Genotox; Citotox; naniparticles




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