Nara Hidenori, Fukano Yasuo, Nishino Tomoki, Aufderheide Michaela
Product Science Division, R&D Group, Japan Tobacco Inc., 6-2, Umegaoka, Aoba-ku, Yokohama, Kanagawa 227-8512, Japan.
Exp Toxicol Pathol. 2013 Jul;65(5):683-8. doi: 10.1016/j.etp.2012.08.004. Epub 2012 Sep 19.
For the biological evaluation of cigarette smoke in vitro, the particulate phase (PP) and the gas vapor phase (GVP) of mainstream smoke have usually been collected individually and exposed to biological material such as cultured cells. Using this traditional method, the GVP is collected by bubbling in an aqueous solution such as phosphate-buffered saline (PBS). In such a way the water-insoluble GVP fraction is excluded from the GVP, meaning that the toxic potential of the water-insoluble GVP fraction has hardly been investigated so far. In our experiments we used a direct exposure method to expose cells at the air-liquid interface (ALI) to the water-insoluble GVP fraction for demonstrating its toxicological/biological activity. In order to isolate the water-insoluble GVP fraction from mainstream smoke, the GVP was passed through 6 impingers connected in series with PBS. After direct exposure of Chinese hamster ovary cells (CHO-K1) with the water-insoluble GVP fraction in the CULTEX(®) system its cytotoxicity was assayed by using the neutral red uptake assay. The water-insoluble GVP fraction was proven to be less cytotoxic than the water-soluble GVP fraction, but showed a significant effect in a dose-dependent manner. The results of this study showed that the direct exposure of cultivated cells at the air-liquid interface offers the possibility to analyze the biological and toxicological activities of all fractions of cigarette smoke including the water-insoluble GVP fraction.
为了在体外对香烟烟雾进行生物学评估，通常会分别收集主流烟雾的颗粒相（PP）和气态挥发相（GVP），并将其暴露于生物材料如培养细胞中。使用这种传统方法时，GVP是通过在诸如磷酸盐缓冲盐水（PBS）的水溶液中鼓泡来收集的。通过这种方式，水不溶性GVP部分被排除在GVP之外，这意味着迄今为止几乎没有研究过水不溶性GVP部分的潜在毒性。在我们的实验中，我们使用直接暴露方法，使气液界面（ALI）处的细胞暴露于水不溶性GVP部分，以证明其毒理学/生物学活性。为了从主流烟雾中分离出水不溶性GVP部分，使GVP通过与PBS串联连接的6个冲击式吸收管。在中国仓鼠卵巢细胞（CHO-K1）在CULTEX®系统中直接暴露于水不溶性GVP部分后，通过中性红摄取试验测定其细胞毒性。结果证明，水不溶性GVP部分的细胞毒性低于水溶性GVP部分，但呈现出显著的剂量依赖性效应。本研究结果表明，在气液界面直接暴露培养细胞，为分析香烟烟雾所有部分（包括水不溶性GVP部分）的生物学和毒理学活性提供了可能。
