Cytotoxic and Transcriptomic Effects in Avian Hepatocytes Exposed to a Complex Mixture from Air Samples, and Their Relation to the Organic Flame Retardant Signature
Assessing complex environmental mixtures and their effects is challenging. In this study, we evaluate the utility of an avian in vitro screening approach to determine the effects of passive air sampler extracts collected from different global megacities on cytotoxicity and gene expression. Concentra...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-11-01
|
| Series: | Toxics |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2305-6304/9/12/324 |
| _version_ | 1797500247710105600 |
|---|---|
| author | Kelsey Ha Pu Xia Doug Crump Amandeep Saini Tom Harner Jason O’Brien |
| author_facet | Kelsey Ha Pu Xia Doug Crump Amandeep Saini Tom Harner Jason O’Brien |
| author_sort | Kelsey Ha |
| collection | DOAJ |
| description | Assessing complex environmental mixtures and their effects is challenging. In this study, we evaluate the utility of an avian in vitro screening approach to determine the effects of passive air sampler extracts collected from different global megacities on cytotoxicity and gene expression. Concentrations of a suite of organic flame retardants (OFRs) were quantified in extracts from a total of 19 megacities/major cities in an earlier study, and levels were highly variable across sites. Chicken embryonic hepatocytes were exposed to serial dilutions of extracts from the 19 cities for 24 h. Cell viability results indicate a high level of variability in cytotoxicity, with extracts from Toronto, Canada, having the lowest LC50 value. Partial least squares (PLS) regression analysis was used to estimate LC50 values from OFR concentrations. PLS modeling of OFRs was moderately predictive of LC50 (<i>p</i>-value = 0.0003, r<sup>2</sup> = 0.66, slope = 0.76, when comparing predicted LC50 to actual values), although only after one outlier city was removed from the analysis. A chicken ToxChip PCR array, comprising 43 target genes, was used to determine effects on gene expression, and similar to results for cell viability, gene expression profiles were highly variable among the megacities. PLS modeling was used to determine if gene expression was related to the OFR profiles of the extracts. Weak relationships to the ToxChip expression profiles could be detected for only three of the 35 OFRs (indicated by regression slopes between 0.6 and 0.5 when comparing predicted to actual OFR concentrations). While this in vitro approach shows promise in terms of evaluating effects of complex mixtures, we also identified several limitations that, if addressed in future studies, might improve its performance. |
| first_indexed | 2024-03-10T03:59:11Z |
| format | Article |
| id | doaj.art-9a36a876c88a419f99d5efa0c654dc7d |
| institution | Directory Open Access Journal |
| issn | 2305-6304 |
| language | English |
| last_indexed | 2024-03-10T03:59:11Z |
| publishDate | 2021-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxics |
| spelling | doaj.art-9a36a876c88a419f99d5efa0c654dc7d2023-11-23T10:49:38ZengMDPI AGToxics2305-63042021-11-0191232410.3390/toxics9120324Cytotoxic and Transcriptomic Effects in Avian Hepatocytes Exposed to a Complex Mixture from Air Samples, and Their Relation to the Organic Flame Retardant SignatureKelsey Ha0Pu Xia1Doug Crump2Amandeep Saini3Tom Harner4Jason O’Brien5Department of Chemistry and Biomolecular Sciences, University of Ottawa, Ottawa, ON K1N 6N5, CanadaNational Wildlife Research Centre, Environment and Climate Change Canada, Ottawa, ON K1A 0H3, CanadaNational Wildlife Research Centre, Environment and Climate Change Canada, Ottawa, ON K1A 0H3, CanadaAir Quality Processes Research Section, Environment and Climate Change Canada, Toronto, ON M3H 5T4, CanadaAir Quality Processes Research Section, Environment and Climate Change Canada, Toronto, ON M3H 5T4, CanadaNational Wildlife Research Centre, Environment and Climate Change Canada, Ottawa, ON K1A 0H3, CanadaAssessing complex environmental mixtures and their effects is challenging. In this study, we evaluate the utility of an avian in vitro screening approach to determine the effects of passive air sampler extracts collected from different global megacities on cytotoxicity and gene expression. Concentrations of a suite of organic flame retardants (OFRs) were quantified in extracts from a total of 19 megacities/major cities in an earlier study, and levels were highly variable across sites. Chicken embryonic hepatocytes were exposed to serial dilutions of extracts from the 19 cities for 24 h. Cell viability results indicate a high level of variability in cytotoxicity, with extracts from Toronto, Canada, having the lowest LC50 value. Partial least squares (PLS) regression analysis was used to estimate LC50 values from OFR concentrations. PLS modeling of OFRs was moderately predictive of LC50 (<i>p</i>-value = 0.0003, r<sup>2</sup> = 0.66, slope = 0.76, when comparing predicted LC50 to actual values), although only after one outlier city was removed from the analysis. A chicken ToxChip PCR array, comprising 43 target genes, was used to determine effects on gene expression, and similar to results for cell viability, gene expression profiles were highly variable among the megacities. PLS modeling was used to determine if gene expression was related to the OFR profiles of the extracts. Weak relationships to the ToxChip expression profiles could be detected for only three of the 35 OFRs (indicated by regression slopes between 0.6 and 0.5 when comparing predicted to actual OFR concentrations). While this in vitro approach shows promise in terms of evaluating effects of complex mixtures, we also identified several limitations that, if addressed in future studies, might improve its performance.https://www.mdpi.com/2305-6304/9/12/324organic flame retardants (OFRs)complex mixturesin vitro screeningPCR arraypassive air samplingchicken embryonic hepatocytes (CEH) |
| spellingShingle | Kelsey Ha Pu Xia Doug Crump Amandeep Saini Tom Harner Jason O’Brien Cytotoxic and Transcriptomic Effects in Avian Hepatocytes Exposed to a Complex Mixture from Air Samples, and Their Relation to the Organic Flame Retardant Signature Toxics organic flame retardants (OFRs) complex mixtures in vitro screening PCR array passive air sampling chicken embryonic hepatocytes (CEH) |
| title | Cytotoxic and Transcriptomic Effects in Avian Hepatocytes Exposed to a Complex Mixture from Air Samples, and Their Relation to the Organic Flame Retardant Signature |
| title_full | Cytotoxic and Transcriptomic Effects in Avian Hepatocytes Exposed to a Complex Mixture from Air Samples, and Their Relation to the Organic Flame Retardant Signature |
| title_fullStr | Cytotoxic and Transcriptomic Effects in Avian Hepatocytes Exposed to a Complex Mixture from Air Samples, and Their Relation to the Organic Flame Retardant Signature |
| title_full_unstemmed | Cytotoxic and Transcriptomic Effects in Avian Hepatocytes Exposed to a Complex Mixture from Air Samples, and Their Relation to the Organic Flame Retardant Signature |
| title_short | Cytotoxic and Transcriptomic Effects in Avian Hepatocytes Exposed to a Complex Mixture from Air Samples, and Their Relation to the Organic Flame Retardant Signature |
| title_sort | cytotoxic and transcriptomic effects in avian hepatocytes exposed to a complex mixture from air samples and their relation to the organic flame retardant signature |
| topic | organic flame retardants (OFRs) complex mixtures in vitro screening PCR array passive air sampling chicken embryonic hepatocytes (CEH) |
| url | https://www.mdpi.com/2305-6304/9/12/324 |
| work_keys_str_mv | AT kelseyha cytotoxicandtranscriptomiceffectsinavianhepatocytesexposedtoacomplexmixturefromairsamplesandtheirrelationtotheorganicflameretardantsignature AT puxia cytotoxicandtranscriptomiceffectsinavianhepatocytesexposedtoacomplexmixturefromairsamplesandtheirrelationtotheorganicflameretardantsignature AT dougcrump cytotoxicandtranscriptomiceffectsinavianhepatocytesexposedtoacomplexmixturefromairsamplesandtheirrelationtotheorganicflameretardantsignature AT amandeepsaini cytotoxicandtranscriptomiceffectsinavianhepatocytesexposedtoacomplexmixturefromairsamplesandtheirrelationtotheorganicflameretardantsignature AT tomharner cytotoxicandtranscriptomiceffectsinavianhepatocytesexposedtoacomplexmixturefromairsamplesandtheirrelationtotheorganicflameretardantsignature AT jasonobrien cytotoxicandtranscriptomiceffectsinavianhepatocytesexposedtoacomplexmixturefromairsamplesandtheirrelationtotheorganicflameretardantsignature |