Japan Tobacco Inc., R&D Group, Yokohama, Japan.
British American Tobacco, R&D, Southampton, UK.
Toxicol Lett. 2019 Oct 15;315:14-22. doi: 10.1016/j.toxlet.2019.08.004. Epub 2019 Aug 7.
In vitro testing can facilitate the rapid assessment of next generation nicotine delivery products (NGPs) with comparisons to combustible tobacco products. In vitro assays for cytotoxicity and oxidative stress were employed at BAT (UK) and JT (Japan) to test total particulate matter (TPM) of a scientific reference cigarette and aerosol collected mass (ACM) of a commercially available E-cigarette and two tobacco heating products (THP). 3R4F TPMs were generated using the Health Canada intense (HCI) regimen, a modified regime (mHCI) for the THP ACMs and the CORESTA recommended method no. 81 for the E-cigarette ACM. Human lung cells were exposed to the test product TPM/ACMs at concentrations between 0-200 μg/ml followed by the employment of commercially available assays for endpoint analysis that included reactive oxygen species (ROS) generation, the glutathione ratio (GSH:GSSG), activation of the antioxidant response elements (ARE) and cellular viability. TPM/ACM nicotine concentrations were quantified using a UPLC-PDA technique. At both laboratories the 3R4F TPM induced significant and dose-dependent responses in all in vitro assays, whereas no significant responses could be measured for the NGP ACMs. In conclusion, both laboratories obtained comparable results across all endpoints therefore demonstrating the utility of the in vitro techniques combined with standardised test products to support the assessment of NGPs.
体外测试可以促进下一代尼古丁输送产品（NGP）的快速评估，并与可燃烟草产品进行比较。英国烟草测试公司（BAT）和日本烟草公司（JT）采用细胞毒性和氧化应激的体外检测方法，对科学参考卷烟的总颗粒物（TPM）和市售电子烟以及两种加热不燃烧烟草产品（THP）的气溶胶收集量（ACM）进行了测试。使用加拿大健康强化（HCI）方案、THP ACM 的改良方案（mHCI）和CORESTA 推荐方法 81 生成了 3R4F TPM，用于电子烟 ACM。在浓度为 0-200μg/ml 的范围内，将测试产品 TPM/ACM 暴露于人类肺细胞中，然后使用市售的终点分析试剂盒进行分析，包括活性氧（ROS）生成、谷胱甘肽比（GSH:GSSG）、抗氧化反应元件（ARE）的激活和细胞活力。使用 UPLC-PDA 技术定量测定 TPM/ACM 中的尼古丁浓度。在两个实验室中，3R4F TPM 在所有体外检测中均引起显著且剂量依赖性的反应，而 NGP ACM 则未测量到显著的反应。总之，两个实验室在所有终点都获得了可比的结果，因此证明了体外技术与标准化测试产品相结合，用于评估 NGP 的有效性。
