In vitro biological assessment of the stability of cigarette smoke aqueous aerosol extracts

Taylor, Mark; Santopietro, Simone; Baxter, Andrew; East, N.; Breheny, Damien; Thorne, David; Gaça, Marianna

doi:10.1186/s13104-020-05337-2

Cited by 8 publications

(13 citation statements)

References 25 publications

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“…The tar components in tobacco and cannabis are similar ( Tashkin, 2013 ), and chemical species of tobacco tar absorb light at 320 nm ( Taylor et al, 2020 ). Thus, to ensure consistency in CaSE preparations between experiments, we standardized each CaSE preparation as previously described for CSE ( Baglole et al, 2008a ; Zago et al, 2013 ; Guerrina et al, 2021a ; Guerrina et al, 2021b ).…”

Section: Resultsmentioning

confidence: 99%

“…Here, we followed the same standardization method for CSE by measuring the absorbance of CaSE at 320 nm, similar to what we have previously used for CSE ( Martey et al, 2004 ; Baglole et al, 2008a ; Zago et al, 2013 ; Zago et al, 2014 ). Because the tar components in tobacco and cannabis are similar ( Tashkin, 2013 ) and the chemical species of tar in tobacco absorb light at 320 nm ( Taylor et al, 2020 ), standardization can be performed via spectroscopy, and confirmation of cannabinoid presence made by a commercial ELISA. One of the limitations of this study is that we measured only Δ 9 -THC and CBD levels in CaSE, and thus cannot provide information on the presence or absence of additional cannabinoids or other compounds, including those could also affect the activity of the CB1 receptor.…”

Section: Discussionmentioning

confidence: 99%

“…CaSE was then filtered using a 0.45-μm filter (25-mm Acrodisc; Pall Corp., Ann Arbor, MI). Because the tar components in tobacco and cannabis are similar ( Tashkin, 2013 ), and chemical species of tobacco tar absorb light at 320 nm ( Taylor et al, 2020 ), we standardized each CaSE preparation as previously described for CSE ( Baglole et al, 2008a ; Zago et al, 2013 ; Guerrina et al, 2021a ; Guerrina et al, 2021b ) to ensure consistency in CaSE preparations between experiments. Similar to CSE preparation described above, an optical density of 0.65 was considered to represent 100% CaSE.…”

Section: Methodsmentioning

confidence: 99%

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Standardized Cannabis Smoke Extract Induces Inflammation in Human Lung Fibroblasts

et al. 2022

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Cannabis (marijuana) is the most commonly used illicit product in the world and is the second most smoked plant after tobacco. There has been a rapid increase in the number of countries legalizing cannabis for both recreational and medicinal purposes. Smoking cannabis in the form of a joint is the most common mode of cannabis consumption. Combustion of cannabis smoke generates many of the same chemicals as tobacco smoke. Although the impact of tobacco smoke on respiratory health is well-known, the consequence of cannabis smoke on the respiratory system and, in particular, the inflammatory response is unclear. Besides the combustion products present in cannabis smoke, cannabis also contains cannabinoids including Δ9-tetrahydrocannabinol (Δ9-THC) and cannabidiol (CBD). These compounds are hydrophobic and not present in aqueous solutions. In order to understand the impact of cannabis smoke on pathological mechanisms associated with adverse respiratory outcomes, the development of in vitro surrogates of cannabis smoke exposure is needed. Therefore, we developed a standardized protocol for the generation of cannabis smoke extract (CaSE) to investigate its effect on cellular mechanisms in vitro. First, we determined the concentration of Δ9-THC, one of the major cannabinoids, by ELISA and found that addition of methanol to the cell culture media during generation of the aqueous smoke extract significantly increased the amount of Δ9-THC. We also observed by LC-MS/MS that CaSE preparation with methanol contains CBD. Using a functional assay in cells for CB1 receptors, the major target of cannabinoids, we found that this CaSE contains Δ9-THC which activates CB1 receptors. Finally, this standardized preparation of CaSE induces an inflammatory response in human lung fibroblasts. This study provides an optimized protocol for aqueous CaSE preparation containing biologically active cannabinoids that can be used for in vitro experimentation of cannabis smoke and its potential impact on various indices of pulmonary health.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Standardized Cannabis Smoke Extract Induces Inflammation in Human Lung Fibroblasts

et al. 2022

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“…However, the technique for generating CSE is also not standardized. 54 In addition, there are no established methods for directly comparing the amount of e-liquid and number of tobacco cigarettes used to create extracts. 25 Consequently, the parameters used here were based on previous literature and e-cig and cigarette user characteristics.…”

Section: Discussionmentioning

confidence: 99%

“…However, there can be batch-tobatch extract variability. 54 Consequently, in future studies, extract preparations should be standardized using spectroscopy methods to ensure minimal variability between preparations. 45 We chose to investigate cytotoxicity, gene expression, and DNA damage following a 24-hour exposure to EVE or CSE.…”

Section: Discussionmentioning

confidence: 99%

Effects of Electronic (E)‐cigarette Vapor and Cigarette Smoke in Cultured Vocal Fold Fibroblasts

et al. 2022

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Objective: The public use of electronic-cigarettes (e-cigs) is rapidly growing. When heated, e-cigs produce a vapor that is inhaled. The vocal folds are among the first tissues exposed to this insult. However, the impact of e-cigs on vocal fold health is almost entirely unknown. Our objective was to evaluate the effects of e-cig vapor on cultured human vocal fold fibroblasts (hVFFs), the primary cell type of the lamina propria. We compared the cellular effects of e-cig vapor without and with nicotine and conventional cigarette smoke.Study Design: In vitro.Methods: E-cig vapor extract (EVE) and cigarette smoke extract (CSE) were created by bubbling vapor and smoke, respectively, into the cell culture medium. hVFFs were exposed to EVE without or with nicotine or CSE for 24 hours. Untreated cells were used as a control group. Cells were harvested, and cytotoxicity, extracellular matrix and inflammatory gene expression, and DNA damage were assessed.Results: Undiluted EVE without and with nicotine reduced the viability of hVFFs to a cytotoxic level. CSE reduced hVFFs viability to a greater extent than EVE and induced DNA damage as measured by DNA double-strand breaks. No changes in gene expression were observed following EVE or CSE exposure.Conclusion: EVE induces cytotoxicity in hVFFs. However, cellular responses were greater following exposure to CSE, suggesting cigarette smoke may induce more harm, at least in the short term. Findings from this investigation improve our understanding of responses of hVFFs to e-cigs and form the basis for an in vitro methodology to study the vocal fold responses to these products.

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Comparative study of the effects of cigarette smoke versus next-generation tobacco and nicotine product extracts on inflammatory biomarkers of human monocytes

Giebe

Brux

Hofmann

et al. 2023

Pflugers Arch - Eur J Physiol

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Monocytes exhibiting a pro-inflammatory phenotype play a key role in adhesion and development of atherosclerotic plaques. As an alternative to smoking, next-generation tobacco and nicotine products (NGP) are now widely used. However, little is known about their pro-inflammatory effects on monocytes. We investigated cell viability, anti-oxidant and pro-inflammatory gene and protein expression in THP-1 monocytes after exposure to aqueous smoke extracts (AqE) of a heated tobacco product (HTP), an electronic cigarette (e-cig), a conventional cigarette (3R4F) and pure nicotine (nic). Treatment with 3R4F reduced cell viability in a dose-dependent manner, whereas exposure to alternative smoking products showed no difference to control. At the highest non-lethal dose of 3R4F (20%), the following notable mRNA expression changes were observed for 3R4F, HTP, and e-cig respectively, relative to control; HMOX1 (6-fold, < 2-fold, < 2-fold), NQO1 (3.5-fold, < 2-fold, < 2-fold), CCL2 (4-fold, 3.5-fold, 2.5-fold), IL1B (4-fold, 3-fold, < 2-fold), IL8 (5-fold, 2-fold, 2-fold), TNF (2-fold, 2-fold, < 2-fold) and ICAM1 was below the 2-fold threshold for all products. With respect to protein expression, IL1B (3-fold, < 2-fold, < 2-fold) and IL8 (3.5-fold, 2-fold, 2-fold) were elevated over the 2-fold threshold, whereas CCL2, TNF, and ICAM1 were below 2-fold expression for all products. At higher doses, greater inductions were observed with all extracts; however, NGP responses were typically lower than 3R4F. In conclusion, anti-oxidative and pro-inflammatory processes were activated by all products. NGPs overall showed lower responses relative to controls than THP-1 cells exposed to 3R4F AqE.

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In vitro biological assessment of the stability of cigarette smoke aqueous aerosol extracts

Cited by 8 publications

References 25 publications

Standardized Cannabis Smoke Extract Induces Inflammation in Human Lung Fibroblasts

Standardized Cannabis Smoke Extract Induces Inflammation in Human Lung Fibroblasts

Effects of Electronic (E)‐cigarette Vapor and Cigarette Smoke in Cultured Vocal Fold Fibroblasts

Comparative study of the effects of cigarette smoke versus next-generation tobacco and nicotine product extracts on inflammatory biomarkers of human monocytes

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