Airborne ultrafine particles in urban areas and workplaces mainly derive from combustion sources (e.g. incomplete diesel fuel combustion, diesel exhaust particles, DEP), abrasion sources (non-exhaust particles, e.g. brake and tire wear particles), or from the unintentional release of engineered nanoparticles (NPs). Metal and metal oxide are among the most relevant NPs emitted by the above mentioned processes, since they are used as fuel additives, are produced by brakes and tires wearing and are among the most globally produced nanomaterials (NMs). The aim of this study was to analyze the combined in vitro effects of DEP and metal oxide NPs (ZnO, CuO) on human lung cells. The human alveolar A549 cells were exposed for 24h to mixtures of standard DEP (NIST 2975) at the subtoxic concentration of 100 μg/ml and increasing concentrations of ZnO and CuO (10, 15, 20, 25 μg/ml). In parallel cells were treated only with the NPs at the same concentrations used in the mixture. After exposure, MTT test and clonogenic assay were used to assess the cytotoxicity. MTT test showed a concentration-dependent decrease in cell viability after exposure to both DEP-ZnO and DEP-CuO mixtures, although ZnO and CuO NPs themselves resulted more cytotoxic than the respective mixtures with DEP. On the contrary, no significant effects were obtained from the clonogenic assay, in term of number of colonies formed after treatment with mixtures, in comparison with the negative control. Additional microscopic analyses are taking place to establish possible NP interference with cells and colonies at morphological level (e.g. cytoskeleton organization, adhesion and migration behavior). These preliminary data suggest that the presence of DEP may introduce new physico-chemical interactions able to reduce the cytotoxicity of ZnO and CuO NPs, likely interfering with their bio-availability and/or bio-reactivity. In conclusion, in the light of providing experimental results reflecting more strictly the environmental conditions, the possible synergistic or antagonistic effects deriving from the mixture of different hazardous airborne particles should be considered, as well as the opportunity to integrate the results with additional toxicity assays and exposure methods. Acknowledgements: EU Horizon 2020 project PROTECT (grant agreement No 720851)
Zerboni, A., Mantecca, P. (2018). Toxicity of DEP and metal oxide nanoparticles mixtures in human lung cells. In ABSTRACTS of the 54th Congress of the European Societies of Toxicology (EUROTOX 2018) (pp.S236-S237) [10.1016/j.toxlet.2018.06.987].
Toxicity of DEP and metal oxide nanoparticles mixtures in human lung cells
Zerboni, A
Primo
;Mantecca, PUltimo
2018
Abstract
Airborne ultrafine particles in urban areas and workplaces mainly derive from combustion sources (e.g. incomplete diesel fuel combustion, diesel exhaust particles, DEP), abrasion sources (non-exhaust particles, e.g. brake and tire wear particles), or from the unintentional release of engineered nanoparticles (NPs). Metal and metal oxide are among the most relevant NPs emitted by the above mentioned processes, since they are used as fuel additives, are produced by brakes and tires wearing and are among the most globally produced nanomaterials (NMs). The aim of this study was to analyze the combined in vitro effects of DEP and metal oxide NPs (ZnO, CuO) on human lung cells. The human alveolar A549 cells were exposed for 24h to mixtures of standard DEP (NIST 2975) at the subtoxic concentration of 100 μg/ml and increasing concentrations of ZnO and CuO (10, 15, 20, 25 μg/ml). In parallel cells were treated only with the NPs at the same concentrations used in the mixture. After exposure, MTT test and clonogenic assay were used to assess the cytotoxicity. MTT test showed a concentration-dependent decrease in cell viability after exposure to both DEP-ZnO and DEP-CuO mixtures, although ZnO and CuO NPs themselves resulted more cytotoxic than the respective mixtures with DEP. On the contrary, no significant effects were obtained from the clonogenic assay, in term of number of colonies formed after treatment with mixtures, in comparison with the negative control. Additional microscopic analyses are taking place to establish possible NP interference with cells and colonies at morphological level (e.g. cytoskeleton organization, adhesion and migration behavior). These preliminary data suggest that the presence of DEP may introduce new physico-chemical interactions able to reduce the cytotoxicity of ZnO and CuO NPs, likely interfering with their bio-availability and/or bio-reactivity. In conclusion, in the light of providing experimental results reflecting more strictly the environmental conditions, the possible synergistic or antagonistic effects deriving from the mixture of different hazardous airborne particles should be considered, as well as the opportunity to integrate the results with additional toxicity assays and exposure methods. Acknowledgements: EU Horizon 2020 project PROTECT (grant agreement No 720851)
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