Rui Zhang1, Myles M Jones2, Ronna E Dornsife3, Tongde Wu4, Vijay Sivaraman2, Robert Tarran5, Rob U Onyenwoke6. 1. Department of Respiratory and Critical Care Medicine, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, PR China; Biomanufacturing Research Institute and Technology Enterprise (BRITE), Durham, NC, United States. 2. Department of Biological and Biomedical Sciences, Durham, NC, United States. 3. Biomanufacturing Research Institute and Technology Enterprise (BRITE), Durham, NC, United States. 4. Biomanufacturing Research Institute and Technology Enterprise (BRITE), Durham, NC, United States; Department of Pharmaceutical Sciences, North Carolina Central University, Durham, NC, United States. 5. Department of Cell Biology & Physiology, University of North Carolina at Chapel Hill, Chapel Hill, NC, United States. 6. Biomanufacturing Research Institute and Technology Enterprise (BRITE), Durham, NC, United States; Department of Pharmaceutical Sciences, North Carolina Central University, Durham, NC, United States. Electronic address: ronyenwo@nccu.edu.
Abstract
RATIONALE: The popularity of new and emerging tobacco products such as E-cigarettes (E-cigs) is rapidly expanding worldwide. However, uncertainties surrounding the potential health consequences due to the use of such products exist and warrant further study. METHODS: Cultured A549 and Calu-3 airway epithelia were exposed to three out of the eight types of JUUL brand e-liquids ("Mint", "Virginia Tobacco" and "Menthol", all containing 3% nicotine at 1% and 3% (vol/vol) dilutions) and assessed for viability using a resazurin-based assay. Intracellular Ca2+ levels were measured using fluorescent indicators and pro-inflammatory cytokine levels were monitored by quantitative PCR (qPCR). Cultures were also analyzed by flow cytometry to evaluate apoptotic markers and cell viability. RESULTS: Exposing the airway epithelial cells to the flavored JUUL e-liquids led to significant cytotoxicity, with the "Mint" flavor being the overall most cytotoxic. The "Mint" flavored e-liquid also led to significant elevations in intracellular Ca2+ and upregulation of the pro-inflammatory cytokine IL-6 and early apoptotic marker Annexin V. CONCLUSIONS: JUUL e-liquid challenge resulted in a loss of airway epithelial cell viability, induced pro-inflammatory responses and eventually caused apoptosis.
RATIONALE: The popularity of new and emerging tobacco products such as E-cigarettes (E-cigs) is rapidly expanding worldwide. However, uncertainties surrounding the potential health consequences due to the use of such products exist and warrant further study. METHODS: Cultured A549 and Calu-3 airway epithelia were exposed to three out of the eight types of JUUL brand e-liquids ("Mint", "Virginia Tobacco" and "Menthol", all containing 3% nicotine at 1% and 3% (vol/vol) dilutions) and assessed for viability using a resazurin-based assay. Intracellular Ca2+ levels were measured using fluorescent indicators and pro-inflammatory cytokine levels were monitored by quantitative PCR (qPCR). Cultures were also analyzed by flow cytometry to evaluate apoptotic markers and cell viability. RESULTS: Exposing the airway epithelial cells to the flavored JUUL e-liquids led to significant cytotoxicity, with the "Mint" flavor being the overall most cytotoxic. The "Mint" flavored e-liquid also led to significant elevations in intracellular Ca2+ and upregulation of the pro-inflammatory cytokine IL-6 and early apoptotic marker Annexin V. CONCLUSIONS: JUUL e-liquid challenge resulted in a loss of airway epithelial cell viability, induced pro-inflammatory responses and eventually caused apoptosis.
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