Exposure to particle debris generated from passenger and truck tires induces different genotoxicity and inflammatory responses in the RAW 264.7 cell line
Autoři:
Anna Poma aff001; Giulia Vecchiotti aff001; Sabrina Colafarina aff001; Osvaldo Zarivi aff001; Lorenzo Arrizza aff002; Piero Di Carlo aff003; Alessandra Di Cola aff005
Působiště autorů:
Department of Life, Health and Environmental Sciences, University of L'Aquila, L'Aquila, Italy
aff001; Center for Microscopy, University of L'Aquila, L'Aquila, Italy
aff002; Department of Psychological, Health and Territorial Sciences, University "G. d'Annunzio" of Chieti-Pescara, Chieti, Italy
aff003; Center of Excellence on Aging and Translational Medicine—Ce.S.I.—Me.T., University "G. d'Annunzio" of Chieti-Pescara, Chieti, Italy
aff004; Tun Abdul Razak Research Centre, Brickendonbury, Hertford, United Kingdom
aff005
Vyšlo v časopise:
PLoS ONE 14(9)
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pone.0222044
Souhrn
A number of studies have shown variable grades of cytotoxicity and genotoxicity in in vitro cell cultures, laboratory animals and humans when directly exposed to particle debris generated from tires. However, no study has compared the effects of particles generated from passenger tires with the effects of particles from truck tires. The aim of this study was to investigate and relate the cyto- and genotoxic effects of different types of particles (PP, passenger tire particles vs. TP, truck tire particles) in vitro using the phagocytic cell line RAW 264.7 (mouse leukaemic monocyte macrophage cell line). The viability of RAW 264.7 cells was determined by the 3- (4,5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H-tetrazolium (MTS) assay following exposure for 4, 24 and 48 hours to different particle concentrations (10 μg / ml, 25 μg / ml, 50 μg / ml, 100 μg / ml). The effects of particles of passenger and truck tires on cell proliferation and genotoxicity were evaluated by means of the cytokinesis-block micronucleus (CBMN) assay following exposure for 24 hours to different particle concentrations (10 μg / ml, 25 μg / ml, 50 μg / ml, 100 μg / ml). In MTS assay, after 24 hours, it was found that PP induced a 30% decrease in metabolic activity at a concentration of 10 μg/ml, while TP caused reductions of 20% and 10% at concentrations of 10 μg/ml and 50 μg/ml, respectively. At 48 hours after the treatments, we observed increased metabolic activity at 50 μg/ml and 100 μg/ml for the PP while only at 50 μg/ml for the TP. The CBMN assay showed a significant increase in the number of micronuclei in the cells incubated with PP in all experimental conditions, while the cells treated with TP showed a meaningful increase only at 10 μg /ml. We utilized the TNF-α ELISA mouse test to detect the production of tumour necrosis factor-alpha (TNF-α) in RAW 264.7 cells. The effect of passenger and truck particles on TNF-α release was evaluated following exposure for 4 and 24 hours.
After 4 hours of incubation, the cells treated with PP and TP at 100 μg / ml showed a slight but significant increase in TNF-α release, while there was a significant increase in the release of TNF-α after 24 hours of incubation with both tire samples in the cells treated with 50 and 100 μg / ml PP. The data obtained show a higher cytotoxic, clastogenic/genotoxic and inflammatory effects of passenger compared to the truck tire particles.
Klíčová slova:
Biology and life sciences – Physical sciences – Chemistry – Research and analysis methods – Developmental biology – Medicine and health sciences – Microbiology – Pathology and laboratory medicine – Polymer chemistry – Macromolecules – Polymers – Materials science – Materials – Physiology – Diagnostic medicine – Signs and symptoms – Immunology – Immune system – Innate immune system – Cytokines – Immune response – Inflammation – Immune physiology – Molecular development – Chemical elements – Ecology and environmental sciences – Pollution – Biological cultures – Cell cultures – Bioassays and physiological analysis – Biochemical analysis – Colorimetric assays – Enzyme assays – Elastomers – Rubber – Zinc – Protozoology – Micronuclei – MTS assay – Air pollution
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