An investigation into E-cigarette cytotoxicity in-vitro using a novel 3D differentiated co-culture model of human airways
An investigation into E-cigarette cytotoxicity in-vitro using a novel 3D differentiated co-culture model of human airways
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DOI：
10.1016/j.tiv.2018.06.020
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发表时间：
2018-10-01
影响因子：
3.2
通讯作者：
Leslie, Laura J.
中科院分区：
文献类型：
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作者：
Bathrinarayanan, Pranav Vasanthi;Brown, James E. P.;Leslie, Laura J.
目前，对于电子烟(ECs)可能对健康造成的不利影响，人们缺乏共识。使用的细胞模型和暴露方法等重要因素决定了EC研究的生理学相关性。本研究旨在利用生理相关的人呼吸道多细胞体外模型评价EC的细胞毒性。人支气管上皮细胞(CALU-3)和肺成纤维细胞(MRC-5)在空气-液体界面共培养11-14天后，使用自制的气雾剂释放系统将其暴露于标准ISO-3308标准的香烟烟雾(WCS)或EC蒸气(ECV)中7m。在较高的暴露时间(1h-6h)下进一步研究ECV的作用。结果表明，WCS在暴露7m后显著降低细胞存活率，ECV仅在暴露时间大于3h时降低细胞存活率。此外，ECV在降低细胞存活率的同时，也增加了IL-6和IL-8的产生。ECV暴露也导致氧化应激显著增加。最后，WCS而不是ECV暴露诱导了caspase 3/7的激活，表明ECV暴露的细胞是caspase非依赖的死亡。总之，我们的结果表明，长时间的ECV暴露(=3h)对促炎介质的产生、氧化应激和细胞活力有显著影响，但对caspase 3/7活性没有影响。
Currently there is a lack of consensus on the possible adverse health effects of E-cigarettes (ECs). Important factors including cell model employed and exposure method determine the physiological relevance of EC studies. The present study aimed to evaluate EC cytotoxicity using a physiologically relevant in-vitro multicellular model of human airways.Human bronchial epithelial cells (CALU-3) and pulmonary fibroblasts (MRC-5) were co-cultured at air-liquidinterface for 11-14 days post which they were exposed to whole cigarette smoke (WCS) or EC vapour (ECV) at standard ISO-3308 regime for 7 m using a bespoke aerosol delivery system. ECV effects were further investigated at higher exposure times (1 h-6 h).Results showed that while WCS significantly reduced cell viability after 7 m, ECV decreased cell viability only at exposure times higher than 3 h. Furthermore, ECV caused elevated IL-6 and IL-8 production despite reduced cell viability. ECV exposure also produced a marked increase in oxidative stress. Finally, WCS but not ECV exposure induced caspase 3/7 activation, suggesting a caspase independent death of ECV exposed cells.Overall, our results indicate that prolonged ECV exposure(>= 3 h) has a significant impact on pro-inflammatory mediators' production, oxidative stress and cell viability but not caspase 3/7 activity.