Particulate matter (PM10) induces in vitro activation of human neutrophils, and lung histopathological alterations in a mouse model

The epidemiological association between exposure to particulate matter (PM₁₀) and various respiratory and cardiovascular problems is well known, but the mechanisms driving these effects remain unclear. Neutrophils play an essential role in immune defense against foreign agents and also participate in the development of inflammatory responses. However, the role of these cells in the PM₁₀ induced inflammatory response is not yet fully established. Thus, this study aims to evaluate the effect of PM₁₀ on the neutrophil-mediated inflammatory response. For this, neutrophils from healthy adult human donors were in vitro exposed to different concentrations of PM₁₀. The cell viability and cytotoxic activity were evaluated by MTT. LDH, propidium iodide and reactive oxygen species (ROS) were quantified by flow cytometry. Interleukin 8 (IL-8) expression, peptidyl arginine deiminase 4 (PAD₄), myeloperoxidase (MPO), and neutrophil elastase (NE) expression were measured by RT-PCR. IL-8 was also quantified by ELISA. Fluorescence microscopy was used to evaluate neutrophil extracellular traps (NETs) release. The in vivo inflammatory responses were assessed in BALB/c mice exposed to PM₁₀ by histopathology and RT-PCR. The analysis shows that PM₁₀ exposure induced a cytotoxic effect on neutrophils, evidenced by necrosis and LDH release at high PM₁₀ concentrations. ROS production, IL-8, MPO, NE expression, and NETs release were increased at all PM₁₀ concentrations assessed. Neutrophil infiltration in bronchoalveolar lavage fluid (BALF), histopathological changes with inflammatory cell infiltration, and CXCL1 expression were observed in PM₁₀-treated mice. The results suggest that lung inflammation in response to PM₁₀ could be mediated by neutrophils activation. In this case, these cells migrate to the lungs and release pro-inflamatory mediators, including ROS, IL-8, and NETs. Thus, contributing to the exacerbation of respiratory pathologies, such as allergies, infectious and obstructive diseases.