Volatile metabolites differentiate air–liquid interface cultures after infection with <i>Staphylococcus aureus</i>

Ahmed, Waqar; Bardin, Emmanuelle; Davis, Michael D.; Sermet‐Gaudelus, Isabelle; Grassin-Delyle, Stanislas; Fowler, Stephen J.

doi:10.1039/d2an01205g

Cited by 9 publications

(4 citation statements)

References 35 publications

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“…It could be worthwhile repeating this model with other cell types, such as endothelial cells or differentiated alveolar cells, and to include immune cells or bacteria to strengthen the robustness of the current findings. Air–liquid interface culture models have previously been used to better mimic the alveolar compartment [ 38 ]. Unfortunately, these models often still involve the use of plastics, increasing the risk of erroneous results through unwanted contaminates [ 16 , 20 ].…”

Section: Discussionmentioning

confidence: 99%

HS-GC–MS analysis of volatile organic compounds after hyperoxia-induced oxidative stress: a validation study

Lilien,

Fenn,

Brinkman

et al. 2024

ICMx

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Background Exhaled volatile organic compounds (VOCs), particularly hydrocarbons from oxidative stress-induced lipid peroxidation, are associated with hyperoxia exposure. However, important heterogeneity amongst identified VOCs and concerns about their precise pathophysiological origins warrant translational studies assessing their validity as a marker of hyperoxia-induced oxidative stress. Therefore, this study sought to examine changes in VOCs previously associated with the oxidative stress response in hyperoxia-exposed lung epithelial cells. Methods A549 alveolar epithelial cells were exposed to hyperoxia for 24 h, or to room air as normoxia controls, or hydrogen peroxide as oxidative-stress positive controls. VOCs were sampled from the headspace, analysed by gas chromatography coupled with mass spectrometry and compared by targeted and untargeted analyses. A secondary analysis of breath samples from a large cohort of critically ill adult patients assessed the association of identified VOCs with clinical oxygen exposure. Results Following cellular hyperoxia exposure, none of the targeted VOCs, previously proposed as breath markers of oxidative stress, were increased, and decane was significantly decreased. Untargeted analysis did not reveal novel identifiable hyperoxia-associated VOCs. Within the clinical cohort, three previously proposed breath markers of oxidative stress, hexane, octane, and decane had no real diagnostic value in discriminating patients exposed to hyperoxia. Conclusions Hyperoxia exposure of alveolar epithelial cells did not result in an increase in identifiable VOCs, whilst VOCs previously linked to oxidative stress were not associated with oxygen exposure in a cohort of critically ill patients. These findings suggest that the pathophysiological origin of previously proposed breath markers of oxidative stress is more complex than just oxidative stress from hyperoxia at the lung epithelial cellular level.

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Section: Discussionmentioning

confidence: 99%

HS-GC–MS analysis of volatile organic compounds after hyperoxia-induced oxidative stress: a validation study

Lilien,

Fenn,

Brinkman

et al. 2024

ICMx

View full text Add to dashboard Cite

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“…However, hexamethyldisiloxane is a silicon-containing compound that is unlikely to be an endogenously produced metabolite. Hexamethyldisiloxane is suggested to originate from various sources, including personal care products such as antiperspirants, cosmetics and hair-care products [39], and may also derive from the sorbent materials used to capture VOCs or from silicone-containing tubing and seals present in VOC capturing equipment [40,41]. These findings suggest that the analysis of breath samples and the identification of a VOC that truly reflects physiological changes must be undertaken with caution.…”

Section: Discussionmentioning

confidence: 99%

Metabolic insights at the finish line: deciphering physiological changes in ultramarathon runners through breath VOC analysis

Chou,

Arthur,

Shaw

et al. 2024

J. Breath Res.

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Exhaustive exercise can induce unique physiological responses in the lungs and other parts of the human body. The volatile organic compounds (VOCs) in exhaled breath are ideal for studying the effects of exhaustive exercise on the lungs due to the proximity of the breath matrix to the respiratory tract. As breath VOCs can originate from the bloodstream, changes in abundance should also indicate broader physiological effects of exhaustive exercise on the body. Currently, there is limited published data on the effects of exhaustive exercise on breath VOCs. Breath has great potential for biomarker analysis as it can be collected non-invasively, and capture real-time metabolic changes to better understand the effects of exhaustive exercise. In this study, we collected breath samples from a small group of elite runners participating in the 2019 Ultra-Trail du Mont Blanc (UTMB) ultra-marathon. The final analysis included matched paired samples collected before and after the race from 24 subjects. All 48 samples were analyzed using the Breath Biopsy Platform with GC-Orbitrap™ via thermal desorption gas chromatography-mass spectrometry (TD-GC-MS). The Wilcoxon signed-rank test was used to determine whether VOC abundances differed between pre- and post-race breath samples (adjusted p < 0.05). We identified a total of 793 VOCs in the breath samples of elite runners. Of these, 63 showed significant differences between pre- and post-race samples after correction for multiple testing (12 decreased, 51 increased). The specific VOCs identified suggest the involvement of fatty acid oxidation, inflammation, and possible altered gut microbiome activity in response to exhaustive exercise. This study demonstrates significant changes in VOC abundance resulting from exhaustive exercise. Further investigation of VOC changes along with other physiological measurements can help improve our understanding of the effect of exhaustive exercise on the body and subsequent differences in VOCs in exhaled breath.

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“…For example, they lack the ability to form tight monolayers of polarized cells, due to their compromised tight junctions (Foster et al 1998 , Elbert et al 1999 ). The use of A549 cells in ALI models is common, with applications ranging from host–pathogen interaction studies to the investigation of epithelial barrier translocation (Bermudez et al 2002 , Hurley et al 2006 , Tamang et al 2012 , Ahmed et al 2023 ). Three-dimensional A549 aggregate models for a P. aeruginosa infection are described by Carterson et al ( 2005 ) and Wang et al ( 2023 ).…”

Section: Cell Selection For Pneumonia Modelsmentioning

confidence: 99%

In vitro modelling of bacterial pneumonia: a comparative analysis of widely applied complex cell culture models

Mahieu,

Van Moll,

De Vooght

et al. 2024

FEMS Microbiology Reviews

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Bacterial pneumonia greatly contributes to the disease burden and mortality of lower respiratory tract infections among all age groups and risk profiles. Therefore, laboratory modelling of bacterial pneumonia remains important for elucidating the complex host-pathogen interactions and to determine drug efficacy and toxicity. In vitro cell culture enables for the creation of high-throughput, specific disease models in a tightly controlled environment. Advanced human cell culture models specifically, can bridge the research gap between the classical two-dimensional cell models and animal models. This review provides an overview of the current status of the development of complex cellular in vitro models to study bacterial pneumonia infections, with a focus on air-liquid interface models, spheroid, organoid, and lung-on-a-chip models. For the wide scale, comparative literature search, we selected six clinically highly relevant bacteria (P. aeruginosa, M. pneumoniae, H. influenzae, M. tuberculosis, S. pneumoniae, and S. aureus). We reviewed the cell lines that are commonly used, as well as trends and discrepancies in the methodology, ranging from cell infection parameters to assay read-outs. We also highlighted the importance of model validation and data transparency in guiding the research field towards more complex infection models.

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Volatile metabolites differentiate air–liquid interface cultures after infection with Staphylococcus aureus

Abstract: Thin film microextraction to sample VOCs from the apical side of an air–liquid interface culture model. After S. aureus infection, infected and uninfected cultures were distinguished using an untargeted metabolomics approach.

Cited by 9 publications

References 35 publications

HS-GC–MS analysis of volatile organic compounds after hyperoxia-induced oxidative stress: a validation study

HS-GC–MS analysis of volatile organic compounds after hyperoxia-induced oxidative stress: a validation study

Metabolic insights at the finish line: deciphering physiological changes in ultramarathon runners through breath VOC analysis

In vitro modelling of bacterial pneumonia: a comparative analysis of widely applied complex cell culture models

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