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Development of a high-throughput in vivo screening platform for particulate matter exposures.

ABSTRACT: Particulate matter (PM) exposure is a public health burden with poorly understood health effect mechanisms and lacking an efficient model to compare the vast diversity of PM exposures. Zebrafish (Danio rerio) are amenable to high-throughput screening (HTS), but few studies have investigated PM toxicity in zebrafish, despite the multitude of advantages. To develop standardized exposure procedures, the urban PM standard reference material (SRM) 1649b was used to systematically determine sample preparation methods, design experimental controls, determine concentration ranges and evaluation procedures. Embryos (n?=?32/treatment) were dechorionated and placed into 96-well plates containing SRM1649b (0-200??g/mL) at 6?h post fertilization (hpf). Developmental toxicity was assessed at 24 and 120 hpf by evaluating morphological changes, embryonic/larval photomotor behavior, and mortality. Differences from blank medium and particle controls were observed for all biological responses measured. Differences due to SRM1649b concentration and preparation method were also observed. Exposure to SRM1649b from DMSO extraction was associated with changes in morphology and mortality and hypoactivity in photomotor responses compared to the DMSO control for the whole particle suspension (76, 68%) and soluble fraction (59, 54%) during the embryonic and larval stages, respectively. Changes in behavioral responses were not observed following exposure to the insoluble fraction of SRM1649b from DMSO extraction. The toxicity bias from PM preparation provided further impetus to select a single HTS exposure method. Based on the biological activity results, the soluble fraction of SRM1649b from DMSO extraction was selected and shown to have concentration dependent cyp1a/GFP expression. This rapid, sensitive and consistently scalable model is a potentially cost-effective vertebrate approach to study the toxicology of PM from diverse locations, and provides a path to identifying the toxic material(s) in these samples, and discover the mechanisms of toxicity.

SUBMITTER: Roper C

PROVIDER: S-EPMC5951187 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature

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Development of a high-throughput in vivo screening platform for particulate matter exposures.

Roper Courtney C Simonich Staci L Massey SLM Tanguay Robert L RL

Environmental pollution (Barking, Essex : 1987) 20180221

Particulate matter (PM) exposure is a public health burden with poorly understood health effect mechanisms and lacking an efficient model to compare the vast diversity of PM exposures. Zebrafish (Danio rerio) are amenable to high-throughput screening (HTS), but few studies have investigated PM toxicity in zebrafish, despite the multitude of advantages. To develop standardized exposure procedures, the urban PM standard reference material (SRM) 1649b was used to systematically determine sample pre ...[more]

PMID: 29751403

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Project description:In this study, we developed a novel method for generating aerosols that are representative of real-world ambient particulate matter (PM) in terms of both physical and chemical characteristics, with the ultimate objective of using them for inhalation exposure studies. The protocol included collection of ambient PM on filters using a high-volume sampler, which were then extracted with ultrapure Milli-Q water using vortexing and sonication. As an alternative approach for collection, ambient particles were directly captured into aqueous slurry samples using the versatile aerosol concentration enrichment system (VACES)/aerosol-into-liquid collector tandem technology. The aqueous samples from both collection protocols were then re-aerosolized using commercially available nebulizers. The physical characteristics (i.e., particle size distribution) of the generated aerosols were examined by the means of a scanning mobility particle sizer (SMPS) connected to a condensation particle counter (CPC) at different compressed air pressures of the nebulizer, and dilution air flow rates. In addition, the collected PM samples (both ambient and re-aerosolized) were chemically analyzed for water-soluble organic carbon (WSOC), elemental and organic carbon (EC/OC), inorganic ions, polycyclic aromatic hydrocarbons (PAHs), and metals and trace elements. Using the aqueous filter extracts, we were able to effectively recover the water-soluble components of ambient PM (e.g., water-soluble organic matter, and water-soluble inorganic ions); however, this method was deficient in recovering some of the important insoluble components such as EC, PAHs, and many of the redox-active trace elements and metals. In contrast, using the VACES/aerosol-into-liquid collector tandem technology for collecting ambient PM directly into water slurry, we were able to preserve the water-soluble and water-insoluble components very effectively. These results illustrate the superiority of the VACES/aerosol-into liquid collector tandem technology to be used in conjunction with the re-aerosolization setup to create aerosols that fully represent ambient PM, making it an attractive choice for application in inhalation exposure studies.

Dataset Information

Development of a high-throughput in vivo screening platform for particulate matter exposures.

Publications

Development of a high-throughput in vivo screening platform for particulate matter exposures.

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