Removal of organic content from diesel exhaust particles alters cellular responses of primary human bronchial epithelial cells cultured at an air-liquid interface

Background Exposure to air pollutants, including diesel particulate matter, has been linked to adverse respiratory health effects. Inhaled diesel particulate matter contains adsorbed organic compounds. It is not clear whether the adsorbed organics or the residual components are more deleterious to airway cells. Using a physiologically relevant model, we investigated the role of diesel organic content on mediating cellular responses of primary human bronchial epithelial cells (HBECs) cultured at an air-liquid interface (ALI). Methods Primary HBECs were cultured and differentiated at ALI for at least 28 days. To determine which component is most harmful, we compared primary HBEC responses elicited by residual (with organics removed) diesel emissions (DE) to those elicited by neat (unmodified) DE for 30 and 60 minutes at ALI, with cigarette smoke condensate (CSC) as the positive control, and filtered air as negative control. Cell viability (WST-1 cell proliferation assay), inflammation (TNF-α, IL-6 and IL-8 ELISA) and changes in gene expression (qRT-PCR for HO-1, CYP1A1, TNF-α and IL-8 mRNA) were measured. Results Immunofluorescence and cytological staining confirmed the mucociliary phenotype of primary HBECs differentiated at ALI. Neat DE caused a comparable reduction in cell viability at 30 or 60 min exposures, whereas residual DE caused a greater reduction at 60 min. When corrected for cell viability, cytokine protein secretion for TNF-α, IL-6 and IL-8 were maximal with residual DE at 60 min. mRNA expression for HO-1, CYP1A1, TNF-α and IL-8 was not significantly different between exposures. Conclusion This study provides new insights into epithelial cell responses to diesel emissions using a physiologically relevant aerosol exposure model. Both the organic content and residual components of diesel emissions play an important role in determining bronchial epithelial cell response in vitro. Future studies should be directed at testing potentially useful interventions against the adverse health effects of air pollution exposure.

Restructuring of carbonaceous particles upon exposure to organic vapours

Recent research has described the restructuring of particles upon exposure to organic vapours; however, as yet hypotheses able to explain this phenomenon are limited. In this study, a range of experiments were performed to explore different hypotheses related to carbonaceous particle restructuring upon exposure to organic and water vapours, such as: the effect of surface tension, the role of organics in flocculating primary particles, as well as the ability of vapours to “wet” the particle surface. The change in mobility diameter (dm) was investigated for a range carbonaceous particle types (diesel exhaust, petrol exhaust, cigarette smoke, candle smoke, particles generated in a heptane/toluene flame, and wood smoke particles) exposed to different organic (heptane, ethanol, and dimethyl sulfoxide/water (1:1 vol%) mixture) and water vapours. Particles were first size-selected and then bubbled through an impinger (bubbler) containing either an organic solvent or water, where particles trapped inside rising bubbles were exposed to saturated vapours of the solvent in the impinger. The size distribution of particles was simultaneously measured upstream and downstream from the impinger. A size-dependent reduction in dm was observed when bubbling diesel exhaust, particles generated in a heptane/toluene flame, and candle smoke particles through heptane, ethanol and a dimethyl sulfoxide/water (1:1 vol %) mixture. In addition, the size distributions of particles bubbled through an impinger were broader. Moreover, an increase of the geometric standard deviation (σ) of the size distributions of particles bubbled through an impinger was also found to be size-dependent. Size-dependent reduction in dm and an increase of σ indicate that particles undergo restructuring to a more compact form, which was confirmed by TEM analysis. However, bubbling of these particles through water did not result in a size-dependent reduction in dm, nor in an increase of σ. Cigarette smoke, petrol exhaust, and wood smoke particles did not result in any substantial change in dm, or σ, when bubbled through organic solvents or water. Therefore, size-dependent reduction in the dm upon bubbling through organic solvents was observed only for particles that had a fractal-like structure, whilst particles that were liquid or were assumed to be spherical did not exhibit any reduction in dm. Compaction of fractal-like particles was attributed to the ability of condensing vapours to efficiently wet the particles. Our results also show that the presence of an organic layer on the surface of fractal-like particles, or the surface tension of the condensed liquid do not influence the extent of compaction.

Performance evaluation of non-thermal plasma on particulate matter, ozone and CO2 correlation for diesel exhaust emission reduction

This study is seeking to investigate the effect of non-thermal plasma technology in the abatement of particulate matter (PM) from the actual diesel exhaust. Ozone (O3) strongly promotes PM oxidation, the main product of which is carbon dioxide (CO2). PM oxidation into the less harmful product (CO2) is the main objective whiles the correlation between PM, O3 and CO2 is considered. A dielectric barrier discharge reactor has been designed with pulsed power technology to produce plasma inside the diesel exhaust. To characterise the system under varied conditions, a range of applied voltages from 11 kVPP to 21kVPP at repetition rates of 2.5, 5, 7.5 and 10 kHz, have been experimentally investigated. The results show that by increasing the applied voltage and repetition rate, higher discharge power and CO2 dissociation can be achieved. The PM removal efficiency of more than 50% has been achieved during the experiments and high concentrations of ozone on the order of a few hundreds of ppm have been observed at high discharge powers. Furthermore, O3, CO2 and PM concentrations at different plasma states have been analysed for time dependence. Based on this analysis, an inverse relationship between ozone concentration and PM removal has been found and the role of ozone in PM removal in plasma treatment of diesel exhaust has been highlighted.

Effect of pulsed power on particle matter in diesel engine exhaust using a DBD plasma reactor

Nonthermal plasma (NTP) treatment of exhaust gas is a promising technology for both nitrogen oxides (NOX) and particulate matter (PM) reduction by introducing plasma into the exhaust gases. This paper considers the effect of NTP on PM mass reduction, PM size distribution, and PM removal efficiency. The experiments are performed on real exhaust gases from a diesel engine. The NTP is generated by applying high-voltage pulses using a pulsed power supply across a dielectric barrier discharge (DBD) reactor. The effects of the applied high-voltage pulses up to 19.44 kVpp with repetition rate of 10 kHz are investigated. In this paper, it is shown that the PM removal and PM size distribution need to be considered both together, as it is possible to achieve high PM removal efficiency with undesirable increase in the number of small particles. Regarding these two important factors, in this paper, 17 kVpp voltage level is determined to be an optimum point for the given configuration. Moreover, particles deposition on the surface of the DBD reactor is found to be a significant phenomenon, which should be considered in all plasma PM removal tests.

Effect of atmospheric ageing on volatility and ROS of biodiesel exhaust nano-particles

Generally, the magnitude of pollutant emissions from diesel engines is ultimately coupled to the structure of fuel molecules. The presence of oxygen, level of unsaturation and the carbon chain length of respective molecules influence the combustion chemistry. It is speculated that increased oxygen content in the fuel may lead to the increased oxidative potential (Stevanovic, S. 2013). Also, upon the exposure to UV and ozone in the atmosphere, the chemical composition of the exhaust is changed. The presence of an oxidant and UV is triggering the cascade of photochemical reactions as well as the partitioning of semi-volatile compounds between the gas and particle phase. To gain an insight into the relationship between the molecular structures of the esters, their volatile organic content and the potential toxicity of diesel exhaust particulate matter, measurements were conducted on a modern common rail diesel engine. This research also investigates the contribution of atmospheric conditions on the transfer of semi-volatile fraction of diesel exhaust from the gas phase to the particle phase and the extent to which semi-volatile compounds (SVOCs) are related to the oxidative potential, expressed through the concentration of reactive oxygen species (ROS) (Stevanovic, S. 2013)...

Reduction of diesel engine exhaust emissions using non-thermal plasma technology

Non-thermal plasma (NTP) is a promising candidate for controlling engine exhaust emissions. Plasma is known as the fourth state of matter, where both electrons and positive ions co-exist. Both gaseous and particle emissions of diesel exhaust undergo chemical changes when they are exposed to plasma. In this project diesel particulate matter (DPM) mitigation from the actual diesel exhaust by using NTP technology has been studied. The effect of plasma, not only on PM mass but also on PM size distribution, physico-chemical structure of PM and PM removal mechanisms, has been investigated. It was found that NTP technology can significantly reduce both PM mass and number. However, under some circumstances particles can be formed by nucleation. Energy required to create the plasma with the current technology is higher than the benchmark set by the commonly used by the automotive industry. Further research will enable the mechanism of particle creation and energy consumption to be optimised.

Influence of biodiesel fuel composition on the morphology and microstructure of particles emitted from diesel engines

This study investigates the morphology, microstructure and surface composition of Diesel engine exhaust particles. The state of agglomeration, the primary particle size and the fractal dimension of exhaust particles from petroleum Diesel (petrodiesel) and biodiesel blends from microalgae, cotton seed and waste cooking oil were investigated by means of high resolution transmission electron microscopy. With primary particle diameters between 12-19 nm, biodiesel blend primary particles are found to be smaller than petrodiesel ones (21±2 nm). Also it was found that soot agglomerates from biodiesels are more compact and spherical, as their fractal dimensions are higher, e.g. 2.2±0.1 for 50% algae biodiesel compared to 1.7±0.1 for petrodiesel. In addition, analysis of the chemical composition by means of x-ray photoelectron spectroscopy revealed an up to a factor of two increased oxygen content on the primary particle surface for biodiesel. The length, curvature and distance of graphene layers were measured showing a greater structural disorder for biodiesel with shorter fringes of higher tortuosity. This change in carbon chemistry may reflect the higher oxygen content of biofuels. Overall, it seems that the oxygen content in the fuels is the underlying reason for the observed morphological change in the resulting soot particles.

A comparative study of the number and mass of fine particles emitted with diesel fuel and marine gas oil (MGO)

The current investigation reports on diesel particulate matter emissions, with special interest in fine particles from the combustion of two base fuels. The base fuels selected were diesel fuel and marine gas oil (MGO). The experiments were conducted with a four-stroke, six-cylinder, direct injection diesel engine. The results showed that the fine particle number emissions measured by both SMPS and ELPI were higher with MGO compared to diesel fuel. It was observed that the fine particle number emissions with the two base fuels were quantitatively different but qualitatively similar. The gravimetric (mass basis) measurement also showed higher total particulate matter (TPM) emissions with the MGO. The smoke emissions, which were part of TPM, were also higher for the MGO. No significant changes in the mass flow rate of fuel and the brake-specific fuel consumption (BSFC) were observed between the two base fuels.

Application of profluorescent nitroxides for measurements of oxidative capacity of combustion generated particles

Oxidative stress caused by generation of free radicals and related reactive oxygen species (ROS) at the sites of deposition has been proposed as a mechanism for many of the adverse health outcomes associated with exposure to particulate matter (PM). Recently, a new profluorescent nitroxide molecular probe (BPEAnit) developed at QUT was applied in an entirely novel, rapid and non-cell based assay for assessing the oxidative potential of particles (i.e. potential of particles to induce oxidative stress). The technique was applied on particles produced by several combustion sources, namely cigarette smoke, diesel exhaust and wood smoke. One of the main findings from the initial studies undertaken at QUT was that the oxidative potential per PM mass significantly varies for different combustion sources as well as the type of fuel used and combustion conditions. However, possibly the most important finding from our studies was that there was a strong correlation between the organic fraction of particles and the oxidative potential measured by the PFN assay, which clearly highlights the importance of organic species in particle-induced toxicity.

Biodiesel with controlled physicochemical properties, a means to further reduce diesel engine particle emissions

The issue of particle emissions from diesel engines is still a matter of concern due its deleterious effects both on human health and environment(Ristovski et al., 2012). Recently, International Agency for Research on Cancer (IARC) inclusion of diesel engine exhaust particles as carcinogenic to human health added a new margin on it. Apart from the use of after treatment technology, biodiesel is also considered as potential way to reduce particle emission alongside with other emissions(Xue, Grift, & Hansen, 2011). Global biodiesel production is still reasonably small compared to its counterpart fossil diesel, but even this small amount comes from a wide variety of feed stocks. Contrary to fossil diesel, the important physicochemical properties of biodiesel vary among different feed stocks(Hoekman, Broch, Robbins, Ceniceros, & Natarajan, 2012).

The application of a profluorescent nitroxide probe to detect reactive oxygen species derived from combustion-generated particulate matter

Particulate pollution has been widely recognised as an important risk factor to human health. In addition to increases in respiratory and cardiovascular morbidity associated with exposure to particulate matter (PM), WHO estimates that urban PM causes 0.8 million premature deaths globally and that 1.5 million people die prematurely from exposure to indoor smoke generated from the combustion of solid fuels. Despite the availability of a huge body of research, the underlying toxicological mechanisms by which particles induce adverse health effects are not yet entirely understood. Oxidative stress caused by generation of free radicals and related reactive oxygen species (ROS) at the sites of deposition has been proposed as a mechanism for many of the adverse health outcomes associated with exposure to PM. In addition to particle-induced generation of ROS in lung tissue cells, several recent studies have shown that particles may also contain ROS. As such, they present a direct cause of oxidative stress and related adverse health effects. Cellular responses to oxidative stress have been widely investigated using various cell exposure assays. However, for a rapid screening of the oxidative potential of PM, less time-consuming and less expensive, cell-free assays are needed. The main aim of this research project was to investigate the application of a novel profluorescent nitroxide probe, synthesised at QUT, as a rapid screening assay in assessing the oxidative potential of PM. Considering that this was the first time that a profluorescent nitroxide probe was applied in investigating the oxidative stress potential of PM, the proof of concept regarding the detection of PM–derived ROS by using such probes needed to be demonstrated and a sampling methodology needed to be developed. Sampling through an impinger containing profluorescent nitroxide solution was chosen as a means of particle collection as it allowed particles to react with the profluorescent nitroxide probe during sampling, avoiding in that way any possible chemical changes resulting from delays between the sampling and the analysis of the PM. Among several profluorescent nitroxide probes available at QUT, bis(phenylethynyl)anthracene-nitroxide (BPEAnit) was found to be the most suitable probe, mainly due to relatively long excitation and emission wavelengths (λex= 430 nm; λem= 485 and 513 nm). These wavelengths are long enough to avoid overlap with the background fluorescence coming from light absorbing compounds which may be present in PM (e.g. polycyclic aromatic hydrocarbons and their derivatives). Given that combustion, in general, is one of the major sources of ambient PM, this project aimed at getting an insight into the oxidative stress potential of combustion-generated PM, namely cigarette smoke, diesel exhaust and wood smoke PM. During the course of this research project, it was demonstrated that the BPEAnit probe based assay is sufficiently sensitive and robust enough to be applied as a rapid screening test for PM-derived ROS detection. Considering that for all three aerosol sources (i.e. cigarette smoke, diesel exhaust and wood smoke) the same assay was applied, the results presented in this thesis allow direct comparison of the oxidative potential measured for all three sources of PM. In summary, it was found that there was a substantial difference between the amounts of ROS per unit of PM mass (ROS concentration) for particles emitted by different combustion sources. For example, particles from cigarette smoke were found to have up to 80 times less ROS per unit of mass than particles produced during logwood combustion. For both diesel and wood combustion it has been demonstrated that the type of fuel significantly affects the oxidative potential of the particles emitted. Similarly, the operating conditions of the combustion source were also found to affect the oxidative potential of particulate emissions. Moreover, this project has demonstrated a strong link between semivolatile (i.e. organic) species and ROS and therefore, clearly highlights the importance of semivolatile species in particle-induced toxicity.

The impact of sulphur content of diesel fuel on ultrafine particle formation

Early this year the Australian Department of Environment and Heritage commissioned a desktop literature review with a focus on ultrafine particles including analysis of health impacts of the particles as well as the impact of sulphur content of diesel fuel on ultrafine particle emission. This paper summarizes the findings of the report on the link between the sulphur content of diesel fuels and the number of ultrafine particles in diesel emissions. The literature search on this topic resulted in over 150 publications. The majority of these publications, although investigating different aspects of the influence of fuel sulphur level on diesel vehicle emissions, were not directly concerned with ultrafine particle emissions. A specific focus of the paper is on: ----- ----- summary of state of knowledge established by the review, and ----- ----- summary of recommendations on the research priorities for Australia to address the information gaps for this issue, and on the appropriate management responses.

Characterisation and source identification of selected pollutants in house dust

House dust is a heterogeneous matrix, which contains a number of biological materials and particulate matter gathered from several sources. It is the accumulation of a number of semi-volatile and non-volatile contaminants. The contaminants are trapped and preserved. Therefore, house dust can be viewed as an archive of both the indoor and outdoor air pollution. There is evidence to show that on average, people tend to stay indoors most of the time and this increases exposure to house dust. The aims of this investigation were to: " assess the levels of Polycyclic Aromatic Hydrocarbons (PAHs), elements and pesticides in the indoor environment of the Brisbane area; " identify and characterise the possible sources of elemental constituents (inorganic elements), PAHs and pesticides by means of Positive Matrix Factorisation (PMF); and " establish the correlations between the levels of indoor air pollutants (PAHs, elements and pesticides) with the external and internal characteristics or attributes of the buildings and indoor activities by means of multivariate data analysis techniques. The dust samples were collected during the period of 2005-2007 from homes located in different suburbs of Brisbane, Ipswich and Toowoomba, in South East Queensland, Australia. A vacuum cleaner fitted with a paper bag was used as a sampler for collecting the house dust. A survey questionnaire was filled by the house residents which contained information about the indoor and outdoor characteristics of their residences. House dust samples were analysed for three different pollutants: Pesticides, Elements and PAHs. The analyses were carried-out for samples of particle size less than 250 µm. The chemical analyses for both pesticides and PAHs were performed using a Gas Chromatography Mass Spectrometry (GC-MS), while elemental analysis was carried-out by using Inductively-Coupled Plasma-Mass Spectroscopy (ICP-MS). The data was subjected to multivariate data analysis techniques such as multi-criteria decision-making procedures, Preference Ranking Organisation Method for Enrichment Evaluations (PROMETHEE), coupled with Geometrical Analysis for Interactive Aid (GAIA) in order to rank the samples and to examine data display. This study showed that compared to the results from previous works, which were carried-out in Australia and overseas, the concentrations of pollutants in house dusts in Brisbane and the surrounding areas were relatively very high. The results of this work also showed significant correlations between some of the physical parameters (types of building material, floor level, distance from industrial areas and major road, and smoking) and the concentrations of pollutants. Types of building materials and the age of houses were found to be two of the primary factors that affect the concentrations of pesticides and elements in house dust. The concentrations of these two types of pollutant appear to be higher in old houses (timber houses) than in the brick ones. In contrast, the concentrations of PAHs were noticed to be higher in brick houses than in the timber ones. Other factors such as floor level, and distance from the main street and industrial area, also affected the concentrations of pollutants in the house dust samples. To apportion the sources and to understand mechanisms of pollutants, Positive Matrix Factorisation (PMF) receptor model was applied. The results showed that there were significant correlations between the degree of concentration of contaminants in house dust and the physical characteristics of houses, such as the age and the type of the house, the distance from the main road and industrial areas, and smoking. Sources of pollutants were identified. For PAHs, the sources were cooking activities, vehicle emissions, smoking, oil fumes, natural gas combustion and traces of diesel exhaust emissions; for pesticides the sources were application of pesticides for controlling termites in buildings and fences, treating indoor furniture and in gardens for controlling pests attacking horticultural and ornamental plants; for elements the sources were soil, cooking, smoking, paints, pesticides, combustion of motor fuels, residual fuel oil, motor vehicle emissions, wearing down of brake linings and industrial activities.

Investigation of the chemical and physical basis of oxidative stress generated by particulate matter using the profluorescent probe technique

Following the growing need for adoption of alternative fuels, this project aimed at getting more information on the oxidative potential of biodiesel particulate matter. Within this scope, the physical and chemical characteristics of biodiesel PM were analysed which lead to identification of reactive organic fractions. An in-house developed proflurescent nitroxide probe was used. This project further developed in-depth understanding of the chemical mechanisms following the detection of the oxidative potential of PM. This knowledge made a significant contribution to our understanding of processes behind negative health effects of pollution and enabled us to further develop new techniques to monitor it.