Impact of after-treatment devices and biofuels on diesel exhausts genotoxicity in A549 cells exposed at air-liquid interface

Barraud, Claire; Corbière, Cécile; Pottier, Ivannah; Estace, E.; Blanchard, Kerry T.; Logie, C.; Lagadu, Stéphanie; Keravec, V.; Pottier, D; Dionnet, Frédéric; Morin, Jean–Paul; Preterre, David; André, Véronique; Monteil, Christelle; Sichel, François

doi:10.1016/j.tiv.2017.04.025

Cited by 19 publications

(8 citation statements)

References 44 publications

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“…The detection of histone H2AX phosphorylation is a commonly used marker of double-strand DNA damage, although γ-H2AX formation is also an early indicator of apoptosis [33]. In previous studies, diesel exhaust particles (DEP) induced double-strand DNA breaks in A549 and BEAS-2B cells [34,35], while no effects of chronic (6 month) exposure of BEAS-2B cells to DEP were found in another report [24]. However, to date, γ-H2AX formation has not been investigated following exposure to complete emissions.…”

Section: Discussionmentioning

confidence: 99%

The Biological Effects of Complete Gasoline Engine Emissions Exposure in a 3D Human Airway Model (MucilAirTM) and in Human Bronchial Epithelial Cells (BEAS-2B)

Rössner

Červená

Vojtíšek-Lom

et al. 2019

IJMS

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The biological effects induced by complete engine emissions in a 3D model of the human airway (MucilAirTM) and in human bronchial epithelial cells (BEAS-2B) grown at the air–liquid interface were compared. The cells were exposed for one or five days to emissions generated by a Euro 5 direct injection spark ignition engine. The general condition of the cells was assessed by the measurement of transepithelial electrical resistance and mucin production. The cytotoxic effects were evaluated by adenylate kinase (AK) and lactate dehydrogenase (LDH) activity. Phosphorylation of histone H2AX was used to detect double-stranded DNA breaks. The expression of the selected 370 relevant genes was analyzed using next-generation sequencing. The exposure had minimal effects on integrity and AK leakage in both cell models. LDH activity and mucin production in BEAS-2B cells significantly increased after longer exposures; DNA breaks were also detected. The exposure affected CYP1A1 and HSPA5 expression in MucilAirTM. There were no effects of this kind observed in BEAS-2B cells; in this system gene expression was rather affected by the time of treatment. The type of cell model was the most important factor modulating gene expression. In summary, the biological effects of complete emissions exposure were weak. In the specific conditions used in this study, the effects observed in BEAS-2B cells were induced by the exposure protocol rather than by emissions and thus this cell line seems to be less suitable for analyses of longer treatment than the 3D model.

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Section: Discussionmentioning

confidence: 99%

The Biological Effects of Complete Gasoline Engine Emissions Exposure in a 3D Human Airway Model (MucilAirTM) and in Human Bronchial Epithelial Cells (BEAS-2B)

Rössner

Červená

Vojtíšek-Lom

et al. 2019

IJMS

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“…The validated HPLC method was used to evaluate the method’s relevance for ATP, ADP, and AMP quantification in a human bronchial epithelial cell model (BEAS-2B). The extraction protocol was based on Barraud et al [ 10 ]. After addition of PCA, the sample was centrifuged to remove proteins and cell debris, and finally, after neutralization with K 2 CO 3 , perchlorates were removed by centrifugation.…”

Section: Resultsmentioning

confidence: 99%

Validation of a Fast and Simple HPLC-UV Method for the Quantification of Adenosine Phosphates in Human Bronchial Epithelial Cells

et al. 2021

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A new HPLC method for the simultaneous quantitative analysis of adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) was developed and validated. ATP, ADP, and AMP were extracted from human bronchial epithelial cells with a rapid extraction procedure and separated with a C18 column (3 × 150 mm, 2.7 µm) using isocratic elution with a mobile phase consisting of 50 mM of potassium hydrogen phosphate (pH 6.80). The absorbance was monitored at 254 nm. The calibration curves were linear in 0.2 to 10 µM, selective, precise, and accurate. This method allowed us to quantify the nucleotides from two cell models: differentiated NHBE primary cells grown at the air–liquid interface (ALI) and BEAS-2B cell line. Our study highlighted the development of a sensitive, simple, and green analytical method that is faster and less expensive than other existing methods to measure ATP, ADP, and AMP and can be carried out on 2D and 3D cell models.

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“…ALI culture conditions allow the differentiation of primary airway epithelial cells under the similar condition as the physiological state. Human lung-derived cells A549 and Calu-3 can also be differentiated under ALI culture and are used as models for toxicological and pharmacological studies with exhausts, nanoparticles and chemical compounds [ [17] , [18] , [19] ]. Here, we applied ALI-cultured A549 cells for experiments with SARS-CoV-2.…”

Section: Discussionmentioning

confidence: 99%

“…In contrast, these cell lines can be cultured and differentiated under ALI conditions. ALI-cultured A549 cells are used for cell-based toxicity assays by inhalation exposure of exhausts, nanoparticles and chemical compounds [ [17] , [18] , [19] ]. A549 cells exert different expression profiles of genes involved in cellular processes such as those in the cell cycle, DNA damage repair and transporters between in submerged and ALI cultures [ 20 , 21 ].…”

Section: Introductionmentioning

confidence: 99%

Air-liquid interphase culture confers SARS-CoV-2 susceptibility to A549 alveolar epithelial cells

Sasaki

Kishimoto

Itakura

et al. 2021

Biochemical and Biophysical Research Communications

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The human lung cell A549 is susceptible to infection with a number of respiratory viruses. However, A549 cells are resistant to Severe Acute Respiratory Syndrome-Coronavirus-2 (SARS-CoV-2) infection in conventional submerged culture, and this would appear to be due to low expression levels of the SARS-CoV-2 entry receptor: angiotensin-converting enzyme-2 (ACE2). Here, we examined SARS-CoV-2 susceptibility to A549 cells after adaptation to air-liquid interface (ALI) culture. A549 cells in ALI culture yielded a layer of mucus on their apical surface, exhibited decreased expression levels of the proliferation marker KI-67 and intriguingly became susceptible to SARS-CoV-2 infection. We found that A549 cells increased the endogenous expression levels of ACE2 and TMPRSS2 following adaptation to ALI culture conditions. Camostat, a TMPRSS2 inhibitor, reduced SARS-CoV-2 infection in ALI-cultured A549 cells. These findings indicate that ALI culture switches the phenotype of A549 cells from resistance to susceptibility to SARS-CoV-2 infection through upregulation of ACE2 and TMPRSS2.

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Impact of after-treatment devices and biofuels on diesel exhausts genotoxicity in A549 cells exposed at air-liquid interface

Cited by 19 publications

References 44 publications

The Biological Effects of Complete Gasoline Engine Emissions Exposure in a 3D Human Airway Model (MucilAirTM) and in Human Bronchial Epithelial Cells (BEAS-2B)

The Biological Effects of Complete Gasoline Engine Emissions Exposure in a 3D Human Airway Model (MucilAirTM) and in Human Bronchial Epithelial Cells (BEAS-2B)

Validation of a Fast and Simple HPLC-UV Method for the Quantification of Adenosine Phosphates in Human Bronchial Epithelial Cells

Air-liquid interphase culture confers SARS-CoV-2 susceptibility to A549 alveolar epithelial cells

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