Combining analytical techniques to assess the translocation of diesel particles across an alveolar tissue barrier in vitro.

BACKGROUND: During inhalation, airborne particles such as particulate matter ≤ 2.5 µm (PM2.5), can deposit and accumulate on the alveolar epithelial tissue. In vivo studies have shown that fractions of PM2.5 can cross the alveolar epithelium to blood circulation, reaching secondary organs beyond the lungs. However, approaches to quantify the translocation of particles across the alveolar epithelium in vivo and in vitro are still not well established. In this study, methods to assess the translocation of standard diesel exhaust particles (DEPs) across permeable polyethylene terephthalate (PET) inserts at 0.4, 1, and 3 µm pore sizes were first optimized with transmission electron microscopy (TEM), ultraviolet-visible spectroscopy (UV-VIS), and lock-in thermography (LIT), which were then applied to study the translocation of DEPs across human alveolar epithelial type II (A549) cells. A549 cells that grew on the membrane (pore size: 3 µm) in inserts were exposed to DEPs at different concentrations from 0 to 80 µg.mL- 1 ( 0 to 44 µg.cm- 2) for 24 h. After exposure, the basal fraction was collected and then analyzed by combining qualitative (TEM) and quantitative (UV-VIS and LIT) techniques to assess the translocated fraction of the DEPs across the alveolar epithelium in vitro. RESULTS: We could detect the translocated fraction of DEPs across the PET membranes with 3 µm pore sizes and without cells by TEM analysis, and determine the percentage of translocation at approximatively 37% by UV-VIS (LOD: 1.92 µg.mL- 1) and 75% by LIT (LOD: 0.20 µg.cm- 2). In the presence of cells, the percentage of DEPs translocation across the alveolar tissue was determined around 1% at 20 and 40 µg.mL- 1 (11 and 22 µg.cm- 2), and no particles were detected at higher and lower concentrations. Interestingly, simultaneous exposure of A549 cells to DEPs and EDTA can increase the translocation of DEPs in the basal fraction. CONCLUSION: We propose a combination of analytical techniques to assess the translocation of DEPs across lung tissues. Our results reveal a low percentage of translocation of DEPs across alveolar epithelial tissue in vitro and they correspond to in vivo findings. The combination approach can be applied to any traffic-generated particles, thus enabling us to understand their involvement in public health.

Human lung cell models to study aerosol delivery - considerations for model design and development.

Human lung tissue models range from simple monolayer cultures to more advanced three-dimensional co-cultures. Each model system can address the interactions of different types of aerosols and the choice of the model and the mode of aerosol exposure depends on the relevant scenario, such as adverse outcomes and endpoints of interest. This review focuses on the functional, as well as structural, aspects of lung tissue from the upper airway to the distal alveolar compartments as this information is relevant for the design of a model as well as how the aerosol properties determine the interfacial properties with the respiratory wall. The most important aspects on how to design lung models are summarized with a focus on (i) choice of appropriate scaffold, (ii) selection of cell types for healthy and diseased lung models, (iii) use of culture condition and assembly, (iv) aerosol exposure methods, and (v) endpoints and verification process. Finally, remaining challenges and future directions in this field are discussed.

In vitro hazard characterization of simulated aircraft cabin bleed-air contamination in lung models using an air-liquid interface (ALI) exposure system.

Contamination of aircraft cabin air can result from leakage of engine oils and hydraulic fluids into bleed air. This may cause adverse health effects in cabin crews and passengers. To realistically mimic inhalation exposure to aircraft cabin bleed-air contaminants, a mini bleed-air contaminants simulator (Mini-BACS) was constructed and connected to an air-liquid interface (ALI) aerosol exposure system (AES). This unique "Mini-BACS + AES" setup provides steady conditions to perform ALI exposure of the mono- and co-culture lung models to fumes from pyrolysis of aircraft engine oils and hydraulic fluids at respectively 200 °C and 350 °C. Meanwhile, physicochemical characteristics of test atmospheres were continuously monitored during the entire ALI exposure, including chemical composition, particle number concentration (PNC) and particles size distribution (PSD). Additional off-line chemical characterization was also performed for the generated fume. We started with submerged exposure to fumes generated from 4 types of engine oil (Fume A, B, C, and D) and 2 types of hydraulic fluid (Fume E and F). Following submerged exposures, Fume E and F as well as Fume A and B exerted the highest toxicity, which were therefore further tested under ALI exposure conditions. ALI exposures reveal that these selected engine oil (0-100 mg/m3) and hydraulic fluid (0-90 mg/m3) fumes at tested dose-ranges can impair epithelial barrier functions, induce cytotoxicity, produce pro-inflammatory responses, and reduce cell viability. Hydraulic fluid fumes are more toxic than engine oil fumes on the mass concentration basis. This may be related to higher abundance of organophosphates (OPs, ≈2800 µg/m3) and smaller particle size (≈50 nm) of hydraulic fluid fumes. Our results suggest that exposure to engine oil and hydraulic fluid fumes can induce considerable lung toxicity, clearly reflecting the potential health risks of contaminated aircraft cabin air.

Optimization of an air-liquid interface in vitro cell co-culture model to estimate the hazard of aerosol exposures.

Inhalation exposure to environmental and occupational aerosol contaminants is associated with many respiratory health problems. To realistically mimic long-term inhalation exposure for toxicity testing, lung epithelial cells need to maintained and exposed under air-liquid interface (ALI) conditions for a prolonged period of time. In addition, to study cellular responses to aerosol particles, lung epithelial cells have to be co-cultured with macrophages. To that aim, we evaluated human bronchial epithelial Calu-3, 16HBE14o- (16HBE), H292, and BEAS-2B cell lines with respect to epithelial morphology, barrier function and cell viability under prolonged ALI culture conditions. Only the Calu-3 cells can retain the monolayer structure and maintain a strong tight junction under long-term ALI culture at least up to 2 weeks. As such, Calu-3 cells were applied as the structural barrier to create co-culture models with human monocyte-derived macrophages (MDMs) and THP-1 derived macrophages (TDMs). Adhesion of macrophages onto the epithelial monolayer was allowed for 4 h with a density of 5 × 104 macrophages/cm2. In comparison to the Calu-3 mono-culture model, Calu-3 + TDM and Calu-3 + MDM co-culture models showed an increased sensitivity in inflammatory responses to lipopolysaccharide (LPS) aerosol at Day 1 of co-culture, with the Calu-3 + MDM model giving a stronger response than Calu-3 + TDM. Therefore, the epithelial monolayer integrity and increased sensitivity make the Calu-3 + MDM co-culture model a preferred option for ALI exposure to inhaled aerosols for toxicity testing.

Comparative toxicity of ultrafine particles around a major airport in human bronchial epithelial (Calu-3) cell model at the air-liquid interface.

Relatively high concentrations of ultrafine particles (UFPs) have been observed around airports, in which aviation and road traffic emissions are the major sources. This raises concerns about the potential health impacts of airport UFPs, particularly in comparison to those emitted by road traffic. UFPs mainly derived from aviation or road traffic emissions were collected from a location near a major international airport, Amsterdam-Schiphol airport (AMS), depending on the wind direction, along with UFPs from an aircraft turbine engine at low and full thrust. Human bronchial epithelial cells (Calu-3) model in combination with an air-liquid interface (ALI) cloud system was used for the in vitro exposure to UFPs at low doses ranging from 0.09 to 2.07 µg/cm2. Particle size distribution was measured. Cell viability, cytotoxicity and inflammatory potential (interleukin (IL) 6 and 8 secretion) on Calu-3 cells were assessed after exposure for 24 h. The biological measurements on Calu-3 cells confirm that pro-inflammatory responses still can be activated at the high cell viability (> 80%) and low cytotoxicity. By the Benchmark Dose (BMD) analysis, Airport and Non-Airport (road traffic) UFPs as well as UFPs samples from a turbine engine have similar toxic properties. Our results suggest that UFPs from aviation and road traffic in airport surroundings may have similar adverse effects on public health.

Organic extract of indoor dust induces estrogen-like effects in human breast cancer cells.

Indoor dust often contains organic contaminants, which adversely impacts human health. In this study, the organic contaminants in the indoor dust from commercial offices and residential houses in Nanjing, China were extracted and their effects on human breast cancer cells (MCF-7) were investigated. Both dust extracts promoted proliferation of MCF-7 cells at ≤24 µg/100 µL, with cell viability being decreased with increasing dust concentrations. Based on LC50, house dust was less toxic than office dust. At 8 µg/100 µL, both extracts caused more MCF-7 cells into active cycling (G2/M + S) and increased intracellular Ca2+ influx, with house dust inducing stronger effects than office dust. Further, the expression of estrogen-responsive genes for TFF1 and EGR3 was enhanced by 3-9 and 4-9 folds, while the expression of cell cycle regulatory genes for cyclin D was enhanced by 2-5 folds. The results suggested that organic dust extract influenced cell viability, altered cell cycle, increased intracellular Ca2+ levels, and activated cell cycle regulatory and estrogen-responsive gene expressions, with house dust showing lower cytotoxicity but higher estrogenic potential on MCF-7 cells. The results indicate the importance of reducing organic contaminants in indoor dust to mitigate their adverse impacts on human health.

An Air-liquid Interface Bronchial Epithelial Model for Realistic, Repeated Inhalation Exposure to Airborne Particles for Toxicity Testing.

For toxicity testing of airborne particles, air-liquid interface (ALI) exposure systems have been developed for in vitro tests in order to mimic realistic exposure conditions. This puts specific demands on the cell culture models. Many cell types are negatively affected by exposure to air (e.g., drying out) and only remain viable for a few days. This limits the exposure conditions that can be used in these models: usually relatively high concentrations are applied as a cloud (i.e., droplets containing particles, which settle down rapidly) within a short period of time. Such experimental conditions do not reflect realistic long-term exposure to low concentrations of particles. To overcome these limitations the use of a human bronchial epithelial cell line, Calu-3 was investigated. These cells can be cultured at ALI conditions for several weeks while retaining a healthy morphology and a stable monolayer with tight junctions. In addition, this bronchial model is suitable for testing the effects of repeated exposures to low, realistic concentrations of airborne particles using an ALI exposure system. This system uses a continuous airflow in contrast to other ALI exposure systems that use a single nebulization producing a cloud. Therefore, the continuous flow system is suitable for repeated and prolonged exposure to airborne particles while continuously monitoring the particle characteristics, exposure concentration, and delivered dose. Taken together, this bronchial model, in combination with the continuous flow exposure system, is able to mimic realistic, repeated inhalation exposure conditions that can be used for toxicity testing.

Differences in cytotoxicity of lung epithelial cells exposed to titanium dioxide nanofibers and nanoparticles: Comparison of air-liquid interface and submerged cell cultures.

Air Liquid Interface (ALI) system has emerged as a useful tool for toxicity evaluation of nanomaterials related to inhalation since the system mimics the aerosol exposure. We compared the biological responses of lung epithelial cells exposed to titanium dioxide (TiO2) nanofibers and nanoparticles in ALI and submerged cell cultures systems. Cells were exposed to 2 and 10 µg/cm2 for 24 h, 48 h and 72 h and LDH release, TiO2 internalization, DNA-double strand breaks (DSBs) and ROS production were assessed. LDH release was similar in both systems and particles had higher cytoplasmic uptake in submerged systems. Both TiO2 types were located in the cytoplasm but nanofibers had nuclear uptake regardless to the system tested. Cells exposed to TiO2 nanofibers had higher DSBs in the ALI system than in submerged cell cultures but cells exposed to TiO2 nanoparticles had similar DSBs in both systems. ROS production was higher in cells exposed to TiO2 nanofibers compared to cells exposed to TiO2 nanoparticles. In conclusion, cytotoxicity of lung epithelial cells was similar in ALI or submerged cell cultures, however cells exposed to TiO2 nanofibers displayed higher toxicity than cells exposed to TiO2 nanoparticles.

The complete chloroplast genome sequence of Actinidia eriantha.

The complete chloroplast (cp) genome sequence of Actinidia eriantha was sequenced and assembled using Illumina paired-end data. The cp genome from A. eriantha is 156,964 bp in length, composed of a pair of 23,892 bp inverted repeat regions (IR) separated by a large single copy region (LSC) of 88,639 bp and a small single copy region (SSC) of 20,541 bp. The cp genome contained 113 unique genes, including 79 protein-coding genes, 30 tRNA genes, and four ribosomal RNA genes. The phylogenetic position of A. eriantha based the cp genome data was sister to the group A. rufa, A. deliciosa, and A. chinensis.

Water extract of indoor dust induces tight junction disruption in normal human corneal epithelial cells.

In corneal epithelium, tight junctions play a vital role in its barrier function. Human cornea is highly susceptible to damage by dust. Continued daily exposure to dust has been associated with increased risks of corneal injury. Studies demonstrated that water extract of dust induced cytotoxicity in human corneal epithelial cells (HCECs); however, its effects on corneal epithelial barrier function are unknown. In this study, we determined the concentrations of heavy metals in water extracts of dust, with office dust having higher concentrations of heavy metals than housedust, and Cu and Zn being highest among metals for both dust. Changes in barrier function and its associated mechanism after exposing HCECs to water extracts of dust at 48 µg/100 µ L for 7 d were evaluated. Water extracts of both dust caused decrease of TEER value (39-73%), down-regulation of gene expression related to tight junction and mucin (0.2-0.8 fold), and loss of ZO-1 immunoreactivity from cellular borders, with office dust having greater potential than housedust to disrupt corneal epithelial barrier function. Our data implied the importance to reduce heavy metals in dust to reduce their adverse impacts on human eyes.

Pro-inflammatory responses to PM0.25 from airport and urban traffic emissions.

Air traffic is rapidly growing, raising concerns about the air pollution in the surroundings of airports and its impact on public health. However, little is known about the impact of air pollution sources on air quality and health in the vicinity of airports. In this study, the sources and adverse health effects of airport-related particulate matter (PM) were investigated and compared to those of urban traffic emissions. Ambient PM0.25 were collected at the Los Angeles International Airport (LAX) and at a central Los Angeles site (USC campus), along with PM2.5 collected directly from turbine and diesel engines. The particle chemical composition, oxidative potential (OP) (ascorbic acid (AA), and electron spin resonance (ESR) assay) as well as their reactive oxygen species (ROS) activity, inflammatory potential (interleukin (IL) 6 and 8 and tumor necrosis factor (TNF)-α) and cytotoxicity on human bronchial epithelial (16HBE) cells were assessed. Chemical composition measurements confirmed that aircraft emissions were the major source to LAX PM0.25, while the sources of the USC samples were more complex, including traffic emissions, suspended road and soil dust, and secondary aerosols. The traffic-related transition metals (Fe and Cu) in LAX and USC samples mainly affected OP values of particles, while multiple factors such as composition, size distribution and internalized amount of particles contributed to the promotion of ROS generation in 16HBE cells during 4 h exposure. Internalized particles in cells might also play an important role in activating inflammatory responses during cell recovery period, with LAX particles being more potent. Our results demonstrated considerable toxicity of airport-related particles, even at low exposure concentrations, suggesting that airport emission as source of PM0.25 may also contribute to the adverse effects on public health attributable to PM. The potency of such particles is in the same range as those collected at a site in urban area impacted heavily by traffic emissions.

Impact of particle size on distribution, bioaccessibility, and cytotoxicity of polycyclic aromatic hydrocarbons in indoor dust.

Contaminants, such as polycyclic aromatic hydrocarbons (PAHs), can be absorbed on the particles of indoor dust, which may pose potential health risks. In this study, indoor dust samples were collected and sieved into 6 size fractions (i.e., <43 µm, 43-63 µm, 63-100 µm, 100-150 µm, 150-200 µm, and 200-2000 µm). Ingestion bioaccessibility of PAHs was measured by physiologically based extraction test. Bioaccessibility in fractions of 200-2000 µm was generally higher than those in other particle sizes. Daily uptake doses based on benzo(a)pyrene toxic equivalency quantity were 1.09-15.0 ng/d/kg, and peaked at fractions of <43 µm, while doses considering bioaccessibility ranged from 0.02 to 0.21 ng/d/kg, and peaked at fractions with relatively larger particle size. Cell toxicity was also investigated by human normal liver cell line viability through exposure to organic extracts of indoor dust fractions with various particle sizes. Our results indicated that it is crucial to consider dust particle size and bioaccessibility during risk assessment.

Cellular responses of normal (HL-7702) and cancerous (HepG2) hepatic cells to dust extract exposure.

Cancerous human liver cell line has been used to test the hepatic toxicity of indoor dust, showing its organic extract decreases cell viability. However, little is known about its impact on normal human liver cell line. In the present study, we compared the cellular responses between carcinoma cell line (HepG2) and normal cell line (HL-7702) after exposing to 10-640 µg/100 µL organic dust extract for 24 h. The dust extract caused cytotoxicity, oxidative damage, inflammatory response and loss of mitochondrial transmembrane potential (MMP) in both cells. The inhibition of cell viability in HL-7702 cells was stronger than that in HepG2 cells, with HL-7702 cells having lower LC50. Higher production of oxidative stress, more loss of MMP and stronger suppression of antioxidant enzymes mRNA level occurred in HepG2 cells, while mRNA expression and hepcidin secretion were enhanced in HL-7702 cells at 40/100 µL, indicating the dust extract probably perturbed their liver Fe homeostasis. Our data showed considerable differences in cellular responses between normal and cancerous cell lines. To obtain accurate data, normal hepatocytes should be employed as they better match with the in vivo tissue than cancerous cell lines.

Impact of particle size on distribution and human exposure of flame retardants in indoor dust.

The effect of dust particle size on the distribution and bioaccessibility of flame retardants (FRs) in indoor dust remains unclear. In this study, we analyzed 20 FRs (including 6 organophosphate flame retardants (OPFRs), 8 polybrominated diphenyl ethers (PBDEs), 4 novel brominated flame retardants (NBFRs), and 2 dechlorane plus (DPs)) in composite dust samples from offices, public microenvironments (PME), and cars in Nanjing, China. Each composite sample (one per microenvironment) was separated into 6 size fractions (F1-F6: 200-2000µm, 150-200µm, 100-150µm, 63-100µm, 43-63µm, and <43µm). FRs concentrations were the highest in car dust, being 16 and 6 times higher than those in offices and PME. The distribution of FRs in different size fractions was Kow-dependent and affected by surface area (Log Kow=1-4), total organic carbon (Log Kow=4-9), and FR migration pathways into dust (Log Kow>9). Bioaccessibility of FRs was measured by the physiologically-based extraction test, with OPFR bioaccessibility being 1.8-82% while bioaccessible PBDEs, NBFRs, and DPs were under detection limits due to their high hydrophobicity. The OPFR bioaccessibility in 200-2000µm fraction was significantly higher than that of <43µm fraction, but with no difference among the other four fractions. Risk assessment was performed for the most abundant OPFR-tris(2-chloroethyl) phosphate. The average daily dose (ADD) values were the highest for the <43µm fraction for all three types of dust using total concentrations, but no consistent trend was found among the three types of dust if based on bioaccessible concentrations. Our results indicated that dust size impacted human exposure estimation of FRs due to their variability in distribution and bioaccessibility among different fractions. For future risk assessment, size selection for dust sampling should be standardized and bioaccessibility of FRs should not be overlooked.

Versatile Cascade Esterification Route to MQ Resins.

We describe a versatile cascade route for manufacturing MQ resins using alkoxysilanes (e.g., tetraethoxysilane (TEOS)) or equivalent oligomers (e.g., ethyl polysilicate (polyTEOS)), a carboxylic acid (typically acetic acid), and hexamethyldisiloxane (MM) as starting materials; a strong acid catalyst is also employed in the one-pot reaction. The siloxane resin synthesis is accompanied by esterification of the carboxylic acid to give ethyl acetate, which acts as an important solvent, making the process more controllable. Contrary to traditional sol-gel methods, no water is introduced in the experiments, but is generated in situ. The strategy offers several advantages, including reproducibility, high yields of siloxane resins with excellent batch-to-batch consistency and without gel formation, narrow dispersity, low Si-hydroxyl residues in the final products, and the ability of increasing the molecular weight by thermal treatment. The process utilizes the green chemistry concepts of lower pollutant formation and higher atom efficiency.

Molecular mechanisms of dust-induced toxicity in human corneal epithelial cells: Water and organic extract of office and house dust.

Human corneal epithelial (HCE) cells are continually exposed to dust in the air, which may cause corneal epithelium damage. Both water and organic soluble contaminants in dust may contribute to cytotoxicity in HCE cells, however, the associated toxicity mechanisms are not fully elucidated. In this study, indoor dust from residential houses and commercial offices in Nanjing, China was collected and the effects of organic and water soluble fraction of dust on primary HCE cells were examined. The concentrations of heavy metals in the dust and dust extracts were determined by ICP-MS and PAHs by GC-MS, with office dust having greater concentrations of heavy metals and PAHs than house dust. Based on LC50, organic extract was more toxic than water extract, and office dust was more toxic than house dust. Accordingly, the organic extracts induced more ROS, malondialdehyde, and 8-Hydroxydeoxyguanosine and higher expression of inflammatory mediators (IL-1ß, IL-6, and IL-8), and AhR inducible genes (CYP1A1, and CYP1B1) than water extracts (p<0.05). Extracts of office dust presented greater suppression of superoxide dismutase and catalase activity than those of house dust. In addition, exposure to dust extracts activated NF-κB signal pathway except water extract of house dust. The results suggested that both water and organic soluble fractions of dust caused cytotoxicity, oxidative damage, inflammatory response, and activation of AhR inducible genes, with organic extracts having higher potential to induce adverse effects on primary HCE cells. The results based on primary HCE cells demonstrated the importance of reducing contaminants in indoor dust to reduce their adverse impacts on human eyes.

Advances in in vitro methods to evaluate oral bioaccessibility of PAHs and PBDEs in environmental matrices.

Cleanup goals for sites contaminated with persistent organic pollutants (POPs) are often established based on total contaminant concentrations. However, mounting evidence suggests that understanding contaminant bioavailability in soils is necessary for accurate assessment of contaminant exposure to humans via oral ingestion pathway. Animal-based in vivo tests have been used to assess contaminant bioavailability in soils; however, due to ethical issues and cost, it is desirable to use in vitro assays as alternatives. Various in vitro methods have been developed, which simulate human gastrointestinal (GI) tract using different digestion fluids. These methods can be used to predict POP bioavailability in soils, foods, and indoor dust after showing good correlation with in vivo animal data. Here, five common in vitro methods are evaluated and compared using PAHs and PBDEs as an example of traditional and emerging POPs. Their applications and limitations are discussed while focusing on method improvements and future challenges to predict POP bioavailability in different matrices. The discussions should shed light for future research to accurately assess human exposure to POPs via oral ingestion pathway.

Mechanisms of housedust-induced toxicity in primary human corneal epithelial cells: Oxidative stress, proinflammatory response and mitochondrial dysfunction.

Human cornea is highly susceptible to damage by dust. Continued daily exposure to housedust has been associated with increasing risks of corneal injury, however, the underlying mechanism has not been elucidated. In this study, a composite housedust sample was tested for its cytotoxicity on primary human corneal epithelial (PHCE) cells, which were exposed to dust at 5-320µg/100µL for 24h. PHCE cell viability showed a concentration-dependent toxic effect, attributing to elevated intracellular ROS. Moreover, when exposed at >20-80µg/100µL, dust-induced oxidative damage was evidenced by increased malondialdehyde and 8-hydroxy-2-deoxyguanosine (1.3-2.3-fold) and decreased antioxidative capacity (1.6-3.5-fold). Alteration of mRNA expression of antioxidant enzymes (SOD1, CAT, HO-1, TRXR1, GSTM1, GSTP1, and GPX1) and pro-inflammatory mediators (IL-1ß, IL-6, IL-8, TNF-α, and MCP-1) were also observed. Furthermore, the mitochondrial transmembrane potential was dissipated from 9.2 to 82%. Our results suggested that dust-induced oxidative stress probably played a vital role in the cytotoxicity in PHCE cells, which may have contributed to dust-induced impairment of human cornea.

Organophosphorus flame retardants and phthalate esters in indoor dust from different microenvironments: Bioaccessibility and risk assessment.

Incidental ingestion of indoor dust is an important pathway for human exposure to organophosphorus flame retardants (OPFRs) and phthalate esters (PAEs). However, little is known about their bioaccessibility in indoor dust. In this study, indoor dust samples were collected from houses, offices, public microenvironments (PMEs), and university dorms, and physiologically based extraction test (PBET) was used to measure the bioaccessibility of OPFRs and PAEs in these dust samples. Total concentrations of OPFRs in dust samples ranged from 0.01 to 63.2 µg g(-1), with significantly lower concentrations in dorm dust (median = 0.30 µg g(-1)) than those in houses (3.12), offices (5.94), and PMEs (11.6). Total PAEs ranged from 5.49 to 2161 µg g(-1) with significantly lower concentrations in dorm dust (379 µg g(-1)) than those in the other three types of dust (767, 515, and 731 µg g(-1)). When subject to PBET, the bioaccessibility of OPFRs ranged from 8.18% (triphenyl phosphate) to 54.5% (Tris(2-chloroisopropyl) phosphate) for OPFRs, and from 1.21% (di-2-ethylhexyl phthalate, DEHP) to 81.1% (dimethyl phthalate) for PAEs. Estimated exposure doses for adults and infants to OPFRs via dust ingestion were much lower than the reference doses (RfD), but intake dose of DEHP for infants was higher than the RfD of 20 µg kg(-1) d(-1). However, the DEHP intake dose did not exceed the RfD after incorporating bioaccessibility into risk assessment. Our data indicated the importance of considering contaminant bioaccessibility during risk assessment of indoor dust.