In Vitro Effects of Particulate Matter Associated with a Wildland Fire in the North-West of Italy

Abstract

Wildland fires, increasing in recent decades in the Mediterranean region due to climate change, can contribute to PM levels and composition. This study aimed to investigate biological effects of PM2.5 (Ø < 2.5 µm) and PM10 (Ø < 10 µm) collected near a fire occurred in the North-West of Italy in 2017 and in three other areas (urban and rural areas). Organic extracts were assessed for mutagenicity using Ames test (TA98 and TA100 strains), cell viability (WST-1 and LDH assays) and genotoxicity (Comet assay) with human bronchial cells (BEAS-2B) and estrogenic activity using a gene reporter assay (MELN cells). In all sites, high levels of PM10 and PM2.5 were measured during the fire suggesting that near and distant sites were influenced by fire pollutants. The PM10 and PM2.5 extracts induced a significant mutagenicity in all sites and the mutagenic effect was increased with respect to historical data. All extracts induced a slight increase of the estrogenic activity but a possible antagonistic activity of PM samples collected near fire was observed. No cytotoxicity or DNA damage was detected. Results confirm that fires could be relevant for human health, since they can worsen the air quality increasing PM concentrations, mutagenic and estrogenic effects.

Biological effects of particulate matter samples during the COVID-19 pandemic: a comparison with the pre-lockdown period in Northwest Italy

“…They were cultured in RPMI 1640 medium supplemented with phenol red, fetal bovine serum (FBS) (10% v/v), l -glutamine (4 mM), and penicillin–streptomycin (100 units/mL–100 µg/mL) (supplemented RPMI medium), at 37 °C and 5% CO 2 . The WST-1 assay was performed as previously described (Gea et al 2021 ). Briefly, cells were seeded in 96-well plates in supplemented RPMI medium without phenol red (5000 cells/well, 100 µL/well) and exposed to PM organic extracts (equivalent to 10, 25, 50 m 3 /mL).…”

“…They were cultured in complete Dulbecco’s Modified Eagle’s Medium Nutrient Mixture F12-Ham supplemented with phenol red, FBS (5% v/v), l -glutamine (4 mM), and penicillin–streptomycin (100 units/mL–100 µg/mL), G418 (1 mg/mL) at 37 °C and 5% CO 2 . The luciferase gene reporter assay was performed as previously described using the One-Glo Luciferase Assay System (Promega) (Balaguer et al 1999 ; Gea et al 2021 ; Schilirò et al 2012 ). Briefly, for 3 days, cells were adapted to the test medium (Dulbecco’s Modified Eagle’s Medium Nutrient Mixture F12-Ham without phenol red, supplemented with l -glutamine (4 mM), penicillin–streptomycin (100 units/mL–100 µg/mL), and 5% v/v of dextran-coated charcoal-treated FBS).…”

“…Moreover, it is hard to define the biological effect of this complex mixture, since antagonistic and synergistic interactions can occur among the chemicals. Therefore, in addition to the quantification of toxic chemicals, the assessment of air quality in terms of toxicity can be performed using effect-based tools (Gea et al 2021 ; Marangon et al 2021 ). These tools are able to quantify the overall biological effect induced by chemical mixtures and are particularly suitable for complex mixtures with low concentrations of multiple chemicals (e.g., PM) (Marangon et al 2021 ).…”

“…Among the different effect-based tools that can be applied to assess the biological effects of air pollutants, the in vitro assays are widely applied also because they provide results in a short period. They are able to quantify different biological effects of PM such as cytotoxicity, genotoxicity, mutagenicity, and hormonal activity, giving insights into the potential health effects induced by different PM samples (Gea et al 2021 ).…”