Ozone and particulate matter co-exposure at the air-liquid interface: Establishing an approach to assess pollutant interactions in vitro
Bibliographic record
Abstract
Air pollution is a complex mixture of gases and particulates that varies spatially and temporally, making attribution of adverse effects to specific individual air pollutants a challenge. To disentangle effects of mixtures in a controlled setting, reproducible and realistic co-exposures of human-relevant models to gaseous and particulate pollutants are needed. Although air-liquid interface (ALI) exposures offer considerable promise as non-animal models for inhalation toxicity testing, a lack of studies comparing individual and co-exposures to gaseous and particulate pollutants has thus far prevented assessment of their strengths and limitations for disentangling effects of pollutant mixtures. Using an integrated ALI exposure system, we characterized the interaction between ozone and particles (25 nm fluorescent polystyrene beads) to assess effects on and reproducibility of critical physical endpoints including temperature, humidity, ozone concentration and particle deposition. Particle deposition and concentration were assessed via three independent methods: fluorescence, quartz crystal microbalance (QCM), and airborne particle count. To evaluate the acute biological effects of an air pollutant mixture in vitro , human lung type 2 epithelial-like cells (A549) were exposed at the ALI to air, ozone (O 3 ), particles, and O 3 + particles (co-exposure) for 1 h ( n = 4 independent repeats/exposure type). Cell injury and inflammation were quantified by extracellular lactate dehydrogenase (LDH) activity and release of proinflammatory cytokines (interleukin (IL)-8 and IL-6) respectively 0 and 24 h post-exposure. Exposures were effective at delivering targeted O 3 exposures under controlled temperature and relative humidity. In-well particle deposition and airborne concentration exiting the exposure system, quantified through parallel methods, were consistent, and increased in relation to aerosolized particle concentration. Levels of each pollutant were effectively maintained in the presence of the other. O 3 alone, and co-exposure to O 3 and particles, increased LDH release from A549 cells, indicating pollutant-specific cytotoxicity. In contrast, IL-8 and IL-6 release (24 h > 0 h) were not changed by exposure to the individual pollutants, but tended to increase following co-exposure. The present work establishes the utility of ALI exposure systems to disentangle individual effects of pollutants from a mixture, and highlights the importance of direct experimental characterization of dosimetry and exposure conditions. • We established air-liquid interface co-exposures to gases and particulates • Key variables such as temperature, humidity and ozone concentration were controlled across exposure scenarios • Determination of particle dose by complementary approaches showed proportionality between aerosolised concentration and deposition • Approach enabled assessment of individual and combined effects of ozone and particles on pulmonary epithelial cells
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".