Effect of Sub-chronic Exposure to Cigarette Smoke, Electronic Cigarette and Waterpipe on Human Lung Epithelial Barrier Function

Overview

Journal BMC Pulm Med

Publisher Biomed Central

Specialty Pulmonary Medicine

Date 2020 Aug 14

PMID 32787821

Citations 18

Authors

Baishakhi Ghosh

Hermes Reyes-Caballero

Sevcan Gul Akgun-Olmez

Kristine Nishida

Lakshmana Chandrala

Lena Smirnova

Shyam Biswal

Venkataramana K Sidhaye

Affiliations

Soon will be listed here.

Abstract

Background: Taking into consideration a recent surge of a lung injury condition associated with electronic cigarette use, we devised an in vitro model of sub-chronic exposure of human bronchial epithelial cells (HBECs) in air-liquid interface, to determine deterioration of epithelial cell barrier from sub-chronic exposure to cigarette smoke (CS), e-cigarette aerosol (EC), and tobacco waterpipe exposures (TW).

Methods: Products analyzed include commercially available e-liquid, with 0% or 1.2% concentration of nicotine, tobacco blend (shisha), and reference-grade cigarette (3R4F). In one set of experiments, HBECs were exposed to EC (0 and 1.2%), CS or control air for 10 days using 1 cigarette/day. In the second set of experiments, exposure of pseudostratified primary epithelial tissue to TW or control air exposure was performed 1-h/day, every other day, until 3 exposures were performed. After 16-18 h of last exposure, we investigated barrier function/structural integrity of the epithelial monolayer with fluorescein isothiocyanate-dextran flux assay (FITC-Dextran), measurements of trans-electrical epithelial resistance (TEER), assessment of the percentage of moving cilia, cilia beat frequency (CBF), cell motion, and quantification of E-cadherin gene expression by reverse-transcription quantitative polymerase chain reaction (RT-qPCR).

Results: When compared to air control, CS increased fluorescence (FITC-Dextran assay) by 5.6 times, whereby CS and EC (1.2%) reduced TEER to 49 and 60% respectively. CS and EC (1.2%) exposure reduced CBF to 62 and 59%, and cilia moving to 47 and 52%, respectively, when compared to control air. CS and EC (1.2%) increased cell velocity compared to air control by 2.5 and 2.6 times, respectively. The expression of E-cadherin reduced to 39% of control air levels by CS exposure shows an insight into a plausible molecular mechanism. Altogether, EC (0%) and TW exposures resulted in more moderate decreases in epithelial integrity, while EC (1.2%) substantially decreased airway epithelial barrier function comparable with CS exposure.

Conclusions: The results support a toxic effect of sub-chronic exposure to EC (1.2%) as evident by disruption of the bronchial epithelial cell barrier integrity, whereas further research is needed to address the molecular mechanism of this observation as well as TW and EC (0%) toxicity in chronic exposures.

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References

Chandrala L, Afshar-Mohajer N, Nishida K, Ronzhes Y, Sidhaye V, Koehler K . A Device for measuring the in-situ response of Human Bronchial Epithelial Cells to airborne environmental agents. Sci Rep. 2019; 9(1):7263. PMC: 6513995. DOI: 10.1038/s41598-019-43784-5. View

Tatsuta M, Kan-O K, Ishii Y, Yamamoto N, Ogawa T, Fukuyama S . Effects of cigarette smoke on barrier function and tight junction proteins in the bronchial epithelium: protective role of cathelicidin LL-37. Respir Res. 2019; 20(1):251. PMC: 6842552. DOI: 10.1186/s12931-019-1226-4. View

Kuehn D, Majeed S, Guedj E, Dulize R, Baumer K, Iskandar A . Impact assessment of repeated exposure of organotypic 3D bronchial and nasal tissue culture models to whole cigarette smoke. J Vis Exp. 2015; (96). PMC: 4354636. DOI: 10.3791/52325. View

Czekala L, Simms L, Stevenson M, Tschierske N, Maione A, Walele T . Toxicological comparison of cigarette smoke and e-cigarette aerosol using a 3D in vitro human respiratory model. Regul Toxicol Pharmacol. 2019; 103:314-324. DOI: 10.1016/j.yrtph.2019.01.036. View

Barber R, Harmer D, Coleman R, Clark B . GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues. Physiol Genomics. 2005; 21(3):389-95. DOI: 10.1152/physiolgenomics.00025.2005. View

Haddad C, Salman R, El-Hellani A, Talih S, Shihadeh A, Saliba N . Reactive Oxygen Species Emissions from Supra- and Sub-Ohm Electronic Cigarettes. J Anal Toxicol. 2018; 43(1):45-50. PMC: 6376456. DOI: 10.1093/jat/bky065. View

Wittekindt O . Tight junctions in pulmonary epithelia during lung inflammation. Pflugers Arch. 2016; 469(1):135-147. PMC: 5203840. DOI: 10.1007/s00424-016-1917-3. View

Li Y, Liu G, Zhang J, Zhong X, He Z . Identification of key genes in human airway epithelial cells in response to respiratory pathogens using microarray analysis. BMC Microbiol. 2018; 18(1):58. PMC: 5994059. DOI: 10.1186/s12866-018-1187-7. View

Khlystov A, Samburova V . Flavoring Compounds Dominate Toxic Aldehyde Production during E-Cigarette Vaping. Environ Sci Technol. 2016; 50(23):13080-13085. DOI: 10.1021/acs.est.6b05145. View

10.

Lee M, LeBouf R, Son Y, Koutrakis P, Christiani D . Nicotine, aerosol particles, carbonyls and volatile organic compounds in tobacco- and menthol-flavored e-cigarettes. Environ Health. 2017; 16(1):42. PMC: 5406907. DOI: 10.1186/s12940-017-0249-x. View

11.

Wyatt T, Schmidt S, Rennard S, Tuma D, Sisson J . Acetaldehyde-stimulated PKC activity in airway epithelial cells treated with smoke extract from normal and smokeless cigarettes. Proc Soc Exp Biol Med. 2000; 225(1):91-7. DOI: 10.1046/j.1525-1373.2000.22511.x. View

12.

Maouche K, Medjber K, Zahm J, Delavoie F, Terryn C, Coraux C . Contribution of α7 nicotinic receptor to airway epithelium dysfunction under nicotine exposure. Proc Natl Acad Sci U S A. 2013; 110(10):4099-104. PMC: 3593882. DOI: 10.1073/pnas.1216939110. View

13.

Voos N, Kaiser L, Mahoney M, Bradizza C, Kozlowski L, Benowitz N . Randomized within-subject trial to evaluate smokers' initial perceptions, subjective effects and nicotine delivery across six vaporized nicotine products. Addiction. 2019; 114(7):1236-1248. PMC: 6646880. DOI: 10.1111/add.14602. View

14.

Hartung T . The Lesser Evil of E-Cigarettes. Sci Am. 2016; 315(2):9. DOI: 10.1038/scientificamerican0816-9. View

15.

Ghosh A, Boucher R, Tarran R . Airway hydration and COPD. Cell Mol Life Sci. 2015; 72(19):3637-52. PMC: 4567929. DOI: 10.1007/s00018-015-1946-7. View

16.

Eapen M, Sharma P, Gaikwad A, Lu W, Myers S, Hansbro P . Epithelial-mesenchymal transition is driven by transcriptional and post transcriptional modulations in COPD: implications for disease progression and new therapeutics. Int J Chron Obstruct Pulmon Dis. 2019; 14:1603-1610. PMC: 6645357. DOI: 10.2147/COPD.S208428. View

17.

Zhou H, Wang X, Brighton L, Hazucha M, Jaspers I, Carson J . Increased nasal epithelial ciliary beat frequency associated with lifestyle tobacco smoke exposure. Inhal Toxicol. 2009; 21(10):875-81. PMC: 2721908. DOI: 10.1080/08958370802555898. View

18.

Xiong R, Wu Q, Muskhelishvili L, Davis K, Shemansky J, Bryant M . Evaluating Mode of Action of Acrolein Toxicity in an In Vitro Human Airway Tissue Model. Toxicol Sci. 2018; 166(2):451-464. DOI: 10.1093/toxsci/kfy226. View

19.

Furubayashi T, Inoue D, Nishiyama N, Tanaka A, Yutani R, Kimura S . Comparison of Various Cell Lines and Three-Dimensional Mucociliary Tissue Model Systems to Estimate Drug Permeability Using an In Vitro Transport Study to Predict Nasal Drug Absorption in Rats. Pharmaceutics. 2020; 12(1). PMC: 7023391. DOI: 10.3390/pharmaceutics12010079. View

20.

Hou W, Hu S, Li C, Ma H, Wang Q, Meng G . Cigarette Smoke Induced Lung Barrier Dysfunction, EMT, and Tissue Remodeling: A Possible Link between COPD and Lung Cancer. Biomed Res Int. 2019; 2019:2025636. PMC: 6613007. DOI: 10.1155/2019/2025636. View