The effects of photochemical aging and interactions with secondary organic aerosols on cellular toxicity of combustion particles.

Fine particulate matter (PM2.5) is associated with numerous adverse health effects, including pulmonary and cardiovascular diseases and premature death. Significant contributors to ambient PM2.5 include combustion particles and secondary organic aerosols (SOA). Combustion particles enter the atmosphere and undergo an aging process that changes their shape and composition, but there is limited study on the health effects of combustion particle aging and interactions with SOA. This study aimed to understand how biological responses to combustion particles would be affected by atmospheric aging and interaction with anthropogenic SOA. Fresh combustion particles underwent photochemical aging in a potential aerosol mass (PAM) oxidation flow reactor and interacted with SOA produced by the oxidation of toluene vapor in the PAM reactor. Photochemical aging and SOA interactions lead to significant changes in the PAH content and oxidative potential of the particle. Photochemical aging and SOA interactions also affected the biological responses, such as the inflammatory response and CYP1A1 induction of the particles in monoculture and coculture cells. These findings highlight the significance of photochemical aging and SOA interactions on the composition and cellular responses of combustion particles.

Role of different mechanisms in pro-inflammatory responses triggered by traffic-derived particulate matter in human bronchiolar epithelial cells.

BACKGROUND: Traffic-derived particles are important contributors to the adverse health effects of ambient particulate matter (PM). In Nordic countries, mineral particles from road pavement and diesel exhaust particles (DEP) are important constituents of traffic-derived PM. In the present study we compared the pro-inflammatory responses of mineral particles and DEP to PM from two road tunnels, and examined the mechanisms involved. METHODS: The pro-inflammatory potential of 100 µg/mL coarse (PM10-2.5), fine (PM2.5-0.18) and ultrafine PM (PM0.18) sampled in two road tunnels paved with different stone materials was assessed in human bronchial epithelial cells (HBEC3-KT), and compared to DEP and particles derived from the respective stone materials. Release of pro-inflammatory cytokines (CXCL8, IL-1α, IL-1ß) was measured by ELISA, while the expression of genes related to inflammation (COX2, CXCL8, IL-1α, IL-1ß, TNF-α), redox responses (HO-1) and metabolism (CYP1A1, CYP1B1, PAI-2) was determined by qPCR. The roles of the aryl hydrocarbon receptor (AhR) and reactive oxygen species (ROS) were examined by treatment with the AhR-inhibitor CH223191 and the anti-oxidant N-acetyl cysteine (NAC). RESULTS: Road tunnel PM caused time-dependent increases in expression of CXCL8, COX2, IL-1α, IL-1ß, TNF-α, COX2, PAI-2, CYP1A1, CYP1B1 and HO-1, with fine PM as more potent than coarse PM at early time-points. The stone particle samples and DEP induced lower cytokine release than all size-fractionated PM samples for one tunnel, and versus fine PM for the other tunnel. CH223191 partially reduced release and expression of IL-1α and CXCL8, and expression of COX2, for fine and coarse PM, depending on tunnel, response and time-point. Whereas expression of CYP1A1 was markedly reduced by CH223191, HO-1 expression was not affected. NAC reduced the release and expression of IL-1α and CXCL8, and COX2 expression, but augmented expression of CYP1A1 and HO-1. CONCLUSIONS: The results indicate that the pro-inflammatory responses of road tunnel PM in HBEC3-KT cells are not attributed to the mineral particles or DEP alone. The pro-inflammatory responses seem to involve AhR-dependent mechanisms, suggesting a role for organic constituents. ROS-mediated mechanisms were also involved, probably through AhR-independent pathways. DEP may be a contributor to the AhR-dependent responses, although other sources may be of importance.

The pro-inflammatory effects of combined exposure to diesel exhaust particles and mineral particles in human bronchial epithelial cells.

BACKGROUND: People are exposed to ambient particulate matter (PM) from multiple sources simultaneously in both environmental and occupational settings. However, combinatory effects of particles from different sources have received little attention in experimental studies. In the present study, the pro-inflammatory effects of combined exposure to diesel exhaust particles (DEP) and mineral particles, two common PM constituents, were explored in human lung epithelial cells. METHODS: Particle-induced secretion of pro-inflammatory cytokines (CXCL8 and IL-1ß) and changes in expression of genes related to inflammation (CXCL8, IL-1α, IL-1ß and COX-2), redox responses (HO-1) and xenobiotic metabolism (CYP1A1 and CYP1B1) were assessed in human bronchial epithelial cells (HBEC3-KT) after combined exposure to different samples of DEP and mineral particles. Combined exposure was also conducted using lipophilic organic extracts of DEP to assess the contribution of soluble organic chemicals. Moreover, the role of the aryl hydrocarbon receptor (AhR) pathway was assessed using an AhR-specific inhibitor (CH223191). RESULTS: Combined exposure to DEP and mineral particles induced increases in pro-inflammatory cytokines and expression of genes related to inflammation and redox responses in HBEC3-KT cells that were greater than either particle sample alone. Moreover, robust increases in the expression of CYP1A1 and CYP1B1 were observed. The effects were most pronounced after combined exposure to α-quartz and DEP from an older fossil diesel, but enhanced responses were also observed using DEP generated from a modern biodiesel blend and several stone particle samples of mixed mineral composition. Moreover, the effect of combined exposure on cytokine secretion could also be induced by lipophilic organic extracts of DEP. Pre-incubation with an AhR-specific inhibitor reduced the particle-induced cytokine responses, suggesting that the effects were at least partially dependent on AhR. CONCLUSIONS: Exposure to DEP and mineral particles in combination induces enhanced pro-inflammatory responses in human bronchial epithelial cells compared with exposure to the individual particle samples. The effects are partly mediated through an AhR-dependent pathway and lipophilic organic chemicals in DEP appear to play a central role. These possible combinatory effects between different sources and components of PM warrant further attention and should also be considered when assessing measures to reduce PM-induced health effects.

Biological effects of combustion-derived particles from different biomass sources on human bronchial epithelial cells.

Combustion-derived particles (CDPs), in particular from traffic, are regarded as a central contributor for adverse health effects linked to air pollution. Recently, also biomass burning has been recognized as an important source for CDPs. Here, the effects of CDPs (PM10) originating from burning of pellet, charcoal and wood on key processes associated to lung carcinogenesis were explored. Human bronchial epithelial cells (HBEC3-KT) were exposed to 2.5 µg/cm2 of CDPs for 24 h and biological effects were examined in terms of cytotoxicity, inflammation, epithelial to mesenchymal transition (EMT)-related effects, DNA damage and genotoxicity. Reduced cell migration, inflammation and modulation of various PM-associated genes were observed mainly after exposure to wood and pellet. In contrast, only particles from pellet burning induced alteration in cell proliferation and DNA damage, which resulted in cell cycle alterations. Charcoal instead, appeared in general less effective in inducing pro-carcinogenic effects. These results illustrate differences in the toxicological profile due to the CDPs source. The different chemical compounds adsorbed on CDPs seemed to be central for particle properties, leading to an activation of various cellular signaling pathways involved in early steps of cancer progression.

Effect of combustion-derived particles on genotoxicity and telomere length: A study on human cells and exposed populations.

Particulate matter (PM) from combustion processes has been associated with oxidative stress to DNA, whereas effects related to telomere dysfunction are less investigated. We collected air-borne PM from a passenger cabin of a diesel-propelled train and at a training facility for smoke diving exercises. Effects on oxidative stress biomarkers, genotoxicity measured by the comet assay and telomere length in PM-exposed A549 cells were compared with the genotoxicity and telomere length in peripheral blood mononuclear cells (PBMCs) from human volunteers exposed to the same aerosol source. Although elevated levels of DNA strand breaks and oxidatively damaged DNA in terms of Fpg-sensitive sites were observed in PBMCs from exposed humans, the PM collected at same locations did not cause genotoxicity in the comet assay in A549 cells. Nevertheless, A549 cells displayed telomere length shortening after four weeks exposure to PM. This is in line with slightly shorter telomere length in PBMCs from exposed humans, although it was not statistically significant. In conclusion, the results indicate that genotoxic potency measured by the comet assay of PM in A549 cells may not predict genotoxicity in exposed humans, whereas telomere length measurements may be a novel indicator of genotoxic stress in cell cultures and humans.

Investigation of particulate matter by FTIR, TEM and elemental analyses in a diesel engine operating on diesel and waste cooking oil-biodiesel.

This study investigated the use of a blend of waste vegetable oil and diesel fuel in a diesel tractor engine by comparing the resulting particulate matter (PM) collected on a filter with the PM of diesel fuel. To this purpose, Fourier transform infrared (FTIR) and elemental analyses were carried out and the PM collected on the filters was examined via scanning electron microscopy (SEM). The study also investigated the overall morphology of soot particles from the blend of 75% waste cooking oil (WCO) + 25% diesel (B75) in a diesel engine at three different loads (75%, 50% and 10%) compared with the morphology of the soot particles from diesel fuel (B0). The FTIR spectra of the B75 fuel exhibited the characteristic regions of oxygen bonds. Compared to the B0 fuel, the increased oxidation from the excess oxygen in the B75 fuel reduced the size and number of soot particles. It was also verified that the degree of unsaturation was related to the oxygen content of the B75 fuel. The FTIR spectra of the B75 fuel showed a C = O tensile band methyl ester in 1734 cm-1 and C-O bands at 1214-1362 cm-1. Furthermore, similar FTIR spectra were seen for the diesel fuel (B0) and the B75 fuel. The results showed a significant reduction in soot particle emissions with the WCO-diesel blend. The results of this research have enabled a better evaluation of the effects of different fuel usage on diesel engine combustion performance and emission characteristics.

Potential role of polycyclic aromatic hydrocarbons as mediators of cardiovascular effects from combustion particles.

Air pollution is the most important environmental risk factor for disease and premature death, and exposure to combustion particles from vehicles is a major contributor. Human epidemiological studies combined with experimental studies strongly suggest that exposure to combustion particles may enhance the risk of cardiovascular disease (CVD), including atherosclerosis, hypertension, thrombosis and myocardial infarction.In this review we hypothesize that adhered organic chemicals like polycyclic aromatic hydrocarbons (PAHs), contribute to development or exacerbation of CVD from combustion particles exposure. We summarize present knowledge from existing human epidemiological and clinical studies as well as experimental studies in animals and relevant in vitro studies. The available evidence suggests that organic compounds attached to these particles are significant triggers of CVD. Furthermore, their effects seem to be mediated at least in part by the aryl hydrocarbon receptor (AhR). The mechanisms include AhR-induced changes in gene expression as well as formation of reactive oxygen species (ROS) and/or reactive electrophilic metabolites. This is in accordance with a role of PAHs, as they seem to be the major chemical group on combustion particles, which bind AhR and/or is metabolically activated by CYP-enzymes. In some experimental models however, it seems as PAHs may induce an inflammatory atherosclerotic plaque phenotype irrespective of DNA- and/or AhR-ligand binding properties. Thus, various components and several signalling mechanisms/pathways are likely involved in CVD induced by combustion particles.We still need to expand our knowledge about the role of PAHs in CVD and in particular the relative importance of the different PAH species. This warrants further studies as enhanced knowledge on this issue may amend risk assessment of CVD caused by combustion particles and selection of efficient measures to reduce the health effects of particular matters (PM).

Effect of collection methods on combustion particle physicochemical properties and their biological response in a human macrophage-like cell line.

In vitro studies are a first step toward understanding the biological effects of combustion-derived particulate matter (cdPM). A vast majority of studies expose cells to cdPM suspensions, which requires a method to collect cdPM and suspend it in an aqueous media. The consequences of different particle collection methods on particle physiochemical properties and resulting biological responses are not fully understood. This study investigated the effect of two common approaches (collection on a filter and a cold plate) and one relatively new (direct bubbling in DI water) approach to particle collection. The three approaches yielded cdPM with differences in particle size distribution, surface area, composition, and oxidative potential. The directly bubbled sample retained the smallest sized particles and the bimodal distribution observed in the gas-phase. The bubbled sample contained ∼50% of its mass as dissolved species and lower molecular weight compounds, not found in the other two samples. These differences in the cdPM properties affected the biological responses in THP-1 cells. The bubbled sample showed greater oxidative potential and cellular reactive oxygen species. The scraped sample induced the greatest TNFα secretion. These findings have implications for in vitro studies of air pollution and for efforts to better understand the underlying mechanisms.

Application of a Quartz Crystal Microbalance to Measure the Mass Concentration of Combustion Particle Suspensions.

Researchers studying the biological effects of combustion particles typically rely on suspending particles in de-ionized (DI) water, buffer, and/or media prior to in vitro or in vivo experiments. However, the hydrophobic nature of combustion particles makes it difficult to obtain well-suspended, evenly dispersed mixtures, which also makes it difficult to obtain equivalent dosing and endpoint comparisons. This study explored the use of a quartz crystal microbalance (QCM) to measure the mass concentration of combustion particle suspensions. It compared the QCM mass concentration to that estimated by placing a known mass of combustion particles in DI water. It also evaluated the effect of drop volume and combustion particle type on QCM measurements. The results showed that QCM is a promising direct method for measuring suspended combustion particle mass concentrations, and it is particularly effective for quantifying concentrations of difficult-to-suspend particles and for combustion particles placed in polystyrene containers, which can lead to substantial particle losses.

Repeated Iron-Soot Exposure and Nose-to-brain Transport of Inhaled Ultrafine Particles.

Particulate exposure has been implicated in the development of a number of neurological maladies such as multiple sclerosis, amyotrophic lateral sclerosis, Alzheimer's disease, and idiopathic Parkinson's disease. Only a few studies have focused on the olfactory pathway as a portal through which combustion-generated particles may enter the brain. The primary objective of this study was to define the deposition, uptake, and transport of inhaled ultrafine iron-soot particles in the nasal cavities of mice to determine whether combustion-generated nanoparticles reach the olfactory bulb via the olfactory epithelium and nerve fascicles. Adult female C57B6 mice were exposed to iron-soot combustion particles at a concentration of 200 µg/m3, which included 40 µg/m3 of iron oxide nanoparticles. Mice were exposed for 6 hr/day, 5 days/week for 5 consecutive weeks (25 total exposure days). Our findings visually demonstrate that inhaled ultrafine iron-soot reached the brain via the olfactory nerves and was associated with indicators of neural inflammation.

Single particle aerosol mass spectrometry of coal combustion particles associated with high lung cancer rates in Xuanwei and Fuyuan, China.

Coal combustion particles (CCPs) are linked to the high incidence of lung cancer in Xuanwei and in Fuyuan, China, but studies on the chemical composition of the CCPs are still limited. Single particle aerosol mass spectrometry (SPAMS) was recently developed to measure the chemical composition and size of single particles in real-time. In this study, SPAMS was used to measure individual combustion particles emitted from Xuanwei and Fuyuan coal samples and the results were compared with those by ICP-MS and transmission electron microscopy (TEM). The total of 38,372 particles mass-analyzed by SPAMS can be divided into 9 groups based on their chemical composition and their number percentages: carbonaceous, Na-rich, K-rich, Al-rich, Fe-rich, Si-rich, Ca-rich, heavy metal-bearing, and PAH-bearing particles. The carbonaceous and PAH-bearing particles are enriched in the size range below 0.56 µm, Fe-bearing particles range from 0.56 to 1.0 µm in size, and heavy metals such as Ti, V, Cr, Cu, Zn, and Pb have diameters below 1 µm. The TEM results show that the particles from Xuanwei and Fuyuan coal combustion can be classified into soot aggregates, Fe-rich particles, heavy metal containing particles, and mineral particles. Non-volatile particles detected by SPAMS could also be observed with TEM. The number percentages by SPAMS also correlate with the mass concentrations measured by ICP-MS. Our results could provide valuable insight for understanding high lung cancer incidence in the area.

Lung inflammation and genotoxicity in mice lungs after pulmonary exposure to candle light combustion particles.

Candle burning produces a large amount of particles that contribute substantially to the exposure to indoor particulate matter. The exposures to various types of combustion particles, such as diesel exhaust particles, have been associated with increased risk of lung cancer by mechanisms that involve oxidative stress, inflammation and genotoxicity. The aim of this study was to compare pulmonary effects of candle light combustion particles (CP) with two benchmark diesel exhaust particles (A-DEP and SRM2975). Intratracheal (i.t.) instillation of CP (5mg/kg bodyweight) in C57BL/6n mice produced a significant influx of alveolar macrophages and polymorphonuclear leukocytes and increased concentrations of proteins and lactate dehydrogenase activity in bronchoalveolar fluid. Lower levels of these markers of inflammation and cytotoxicity were observed after i.t. instillation of the same dose of A-DEP or SRM2975. The i.t. instillation of CP did not generate oxidative damage to DNA in lung tissue, measured as DNA strand breaks and human 8-oxoguanine glycosylase-sensitive sites by the comet assay. The lack of genotoxic response was confirmed in lung epithelial (A549) cells, although the exposure to CP increased intracellular levels of reactive oxygen species. In conclusion, pulmonary exposure to particles from burning candles is associated with inflammation and cytotoxicity in the lungs.

Performance of N95 FFRs Against Combustion and NaCl Aerosols in Dry and Moderately Humid Air: Manikin-based Study.

OBJECTIVES: The first objective of this study was to evaluate the penetration of particles generated from combustion of plastic through National Institute for Occupational Safety and Health (NIOSH)-certified N95 filtering facepiece respirators (FFRs) using a manikin-based protocol and compare the data to the penetration of NaCl particles. The second objective was to investigate the effect of relative humidity (RH) on the filtration performance of N95 FFRs. METHODS: Two NIOSH-certified N95 FFRs (A and B) were fully sealed on a manikin headform and challenged with particles generated by combustion of plastic and NaCl particles. The tests were performed using two cyclic flows [with mean inspiratory flow (MIF) rates = 30 and 85 l min(-1), representing human breathing under low and moderate workload conditions] and two RH levels (≈20 and ≈80%, representing dry and moderately humid air). The total and size-specific particle concentrations inside (C in) and outside (C out) of the respirators were measured with a condensation particle counter and an aerosol size spectrometer. The penetration values (C in/C out) were calculated after each test. RESULTS: The challenge aerosol, RH, MIF rate, and respirator type had significant (P < 0.05) effects on the performance of the manikin-sealed FFR. Its efficiency significantly decreased when the FFR was tested with plastic combustion particles compared to NaCl aerosols. For example, at RH ≈80% and MIF = 85 l min(-1), as much as 7.03 and 8.61% of combustion particles penetrated N95 respirators A and B, respectively. The plastic combustion particles and gaseous compounds generated by combustion likely degraded the electric charges on fibers, which increased the particle penetration. Increasing breathing flow rate or humidity increased the penetration (reduced the respirator efficiency) for all tested aerosols. The effect of particle size on the penetration varied depending on the challenge aerosol and respirator type. It was observed that the peak of the size distribution of combustion particles almost coincided with their most penetrating particle size, which was not the case for NaCl particles. This finding was utilized for the data interpretation. CONCLUSIONS: N95 FFRs have lower filter efficiency when challenged with contaminant particles generated by combustion, particularly when used under high humidity conditions compared to NaCl particles.

Short-term dynamics of indoor and outdoor endotoxin exposure: Case of Santiago, Chile, 2012.

Indoor and outdoor endotoxin in PM2.5 was measured for the very first time in Santiago, Chile, in spring 2012. Average endotoxin concentrations were 0.099 and 0.094 [EU/m(3)] for indoor (N=44) and outdoor (N=41) samples, respectively; the indoor-outdoor correlation (log-transformed concentrations) was low: R=-0.06, 95% CI: (-0.35 to 0.24), likely owing to outdoor spatial variability. A linear regression model explained 68% of variability in outdoor endotoxins, using as predictors elemental carbon (a proxy of traffic emissions), chlorine (a tracer of marine air masses reaching the city) and relative humidity (a modulator of surface emissions of dust, vegetation and garbage debris). In this study, for the first time a potential source contribution function (PSCF) was applied to outdoor endotoxin measurements. Wind trajectory analysis identified upwind agricultural sources as contributors to the short-term, outdoor endotoxin variability. Our results confirm an association between combustion particles from traffic and outdoor endotoxin concentrations. For indoor endotoxins, a predictive model was developed but it only explained 44% of endotoxin variability; the significant predictors were tracers of indoor PM2.5 dust (Si, Ca), number of external windows and number of hours with internal doors open. Results suggest that short-term indoor endotoxin variability may be driven by household dust/garbage production and handling. This would explain the modest predictive performance of published models that use answers to household surveys as predictors. One feasible alternative is to increase the sampling period so that household features would arise as significant predictors of long-term airborne endotoxin levels.

Soot and house dust mite allergen cause eosinophilic laryngitis in an animal model.

OBJECTIVES/HYPOTHESIS: Chronic laryngitis (CL) is common and costly. One of the most common causes of CL is thought to be laryngopharyngeal reflux, although a significant percentage of individuals fail to get better with acid suppressive therapy. The role of other potential causes of CL such as allergy and environmental pollution has not been thoroughly investigated. PURPOSE: To evaluate the association between iron soot, house dust mite allergen (HDMA), and CL in an established animal model. METHODS: Twenty-four guinea pigs were separated into four 6-week exposure groups: 1) saline (allergen control) + filtered air (pollution control); 2) HDMA (Dermatophygoides farinae) + filtered air; 3) saline + combustion particulates; or 4) HDMA + combustion particulates. The primary outcome measure was mean eosinophil profile (MEP) in glottic, subglottic, and trachea epithelium and submucosa. RESULTS: The combination of iron soot and HDMA caused eosinophilia (elevated MEP) in the glottic (P < 0.06), subglottic (P < 0.05), and trachea (P < 0.05) submucosa and epithelium (P < 0.05). CONCLUSION: The combination of HDMA and iron soot resulted in laryngeal eosinophilia in an established guinea pig model of CL. The data support the notion that factors other than reflux may cause CL. Further investigation into eosinophilic laryngitis as a distinct clinical entity caused by exposure to environmental allergen and pollution is warranted.

Penetration of Combustion Aerosol Particles Through Filters of NIOSH-Certified Filtering Facepiece Respirators (FFRs).

Filtering facepiece respirators (FFRs) are commonly worn by first responders, first receivers, and other exposed groups to protect against exposure to airborne particles, including those originated by combustion. Most of these FFRs are NIOSH-certified (e.g., N95-type) based on the performance testing of their filters against charge-equilibrated aerosol challenges, e.g., NaCl. However, it has not been examined if the filtration data obtained with the NaCl-challenged FFR filters adequately represent the protection against real aerosol hazards such as combustion particles. A filter sample of N95 FFR mounted on a specially designed holder was challenged with NaCl particles and three combustion aerosols generated in a test chamber by burning wood, paper, and plastic. The concentrations upstream (Cup) and downstream (Cdown) of the filter were measured with a TSI P-Trak condensation particle counter and a Grimm Nanocheck particle spectrometer. Penetration was determined as (Cdown/Cup) ×100%. Four test conditions were chosen to represent inhalation flows of 15, 30, 55, and 85 L/min. Results showed that the penetration values of combustion particles were significantly higher than those of the "model" NaCl particles (p < 0.05), raising a concern about applicability of the N95 filters performance obtained with the NaCl aerosol challenge to protection against combustion particles. Aerosol type, inhalation flow rate and particle size were significant (p < 0.05) factors affecting the performance of the N95 FFR filter. In contrast to N95 filters, the penetration of combustion particles through R95 and P95 FFR filters (were tested in addition to N95) were not significantly higher than that obtained with NaCl particles. The findings were attributed to several effects, including the degradation of an N95 filter due to hydrophobic organic components generated into the air by combustion. Their interaction with fibers is anticipated to be similar to those involving "oily" particles. The findings of this study suggest that the efficiency of N95 respirator filters obtained with the NaCl aerosol challenge may not accurately predict (and rather overestimate) the filter efficiency against combustion particles.