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Culturing of Human Nasal Epithelial Cells at the Air Liquid Interface
Published on: October 8, 2013
Winter fine particulate matter from Milan induces morphological and functional alterations in human pulmonary
Maurizio Gualtieri1, Paride Mantecca, Viviana Corvaja
1POLARIS Research Center, Department of Environmental Sciences, University of Milano Bicocca, Milan, Italy.
Abstract:
Samples of PM(2.5) were gravimetrically collected during the winter 2005/2006 in the urban area of Milan (North Italy). Samples were chemically characterized and the particles were detached from filters to determine their cytotoxic effects on the A549 cell line. Based on the potential toxicological relevance of its components, Milan winter PM(2.5) contained high concentrations of pro-oxidant transition metals and PAHs, while re-suspended particles showed a relatively high frequency of dimensional classes ranging from 40 nm to 300 nm. A549 cells exposed to particle suspensions showed a concentration-dependent decrease in viability, starting from 10 microg/cm(2). Phagocytosis of particles by A549 cells and particle aggregates were morphologically characterized and seemed to depend on both particle concentration and exposure time, with the majority of particles being engulfed in membrane-bound vacuoles after 24h of exposure. The ability of ultrafine particles to penetrate and spread throughout the cells was also verified. Cell membrane lysis and mitochondrial ultrastructural disruption appeared to be the main modifications induced by PM(2.5) on A549 cells. Concomitantly to the adverse effects observed in terms of cell mortality and ultrastructural lesions, a significant intracellular production of reactive oxygen species (ROS) was observed, suggesting that the cytotoxicity, exerted by the winter PM(2.5) in Milan, derived also from its oxidative potential, probably associated with particle-adsorbed metals and PAHs.
Insights
Milan
Area of Science:
- Environmental Science
- Toxicology
- Cell Biology
Background:
- Particulate Matter (PM2.5) in urban environments poses significant health risks.
- Winter PM2.5 in Milan is characterized by high levels of transition metals and polycyclic aromatic hydrocarbons (PAHs).
- Ultrafine particles exhibit unique toxicological properties due to their size and composition.
Purpose of the Study:
- To investigate the cytotoxic effects of Milan winter PM2.5 on A549 lung cells.
- To characterize the physical and chemical properties of PM2.5 relevant to toxicity.
- To elucidate the mechanisms underlying PM2.5-induced cellular damage.
Main Methods:
- Gravimetric collection and chemical characterization of PM2.5 samples.
- Detachment of particles from filters for cell exposure studies.
- Assessment of cell viability, phagocytosis, particle penetration, and ultrastructural changes in A549 cells.
- Measurement of intracellular reactive oxygen species (ROS) production.
Main Results:
- PM2.5 exposure caused a concentration-dependent decrease in A549 cell viability.
- Particles were phagocytosed by A549 cells, with uptake dependent on concentration and time.
- Cell membrane lysis and mitochondrial damage were observed, indicating significant cellular injury.
- Increased intracellular ROS production was linked to PM2.5 cytotoxicity, suggesting oxidative stress.
Conclusions:
- Milan winter PM2.5 exhibits significant cytotoxicity towards A549 cells.
- The observed toxicity is mediated by oxidative stress, likely driven by transition metals and PAHs.
- Ultrafine particle characteristics, including size and composition, are critical factors in PM2.5-induced lung cell damage.
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