Aufderheide Michaela, Knebel Jan W, Ritter Detlef
Department of In Vitro Toxicology, Fraunhofer Institute of Toxicology and Experimental Medicine, Hannover, Germany.
Exp Toxicol Pathol. 2003 Jul;55(1):51-7. doi: 10.1078/0940-2993-00298.
Numerous approaches have been employed for testing the biological activity of cigarette smoke in vitro. None of them has managed to expose cultured lung cells in a realistic manner to the complex gaseous and particulate mixture that constitutes cigarette smoke. We have devised a system that makes this possible. The system presented here enables the direct exposure of human lung cells to native, unmodified cigarette mainstream smoke. It consists of a smoking machine, a dilution device for the smoke, analytical devices for online monitoring and a specially adapted exposure module based on the Cultex** cell cultivation system that is equipped with a gas-exposure top. Due to the special design of the exposure device and the optimised exposure conditions, this equipment allows cultured human lung cells to be exposed to freshly generated cigarette mainstream smoke. Exploratory experiments revealed that the smoke could be diluted over a wide concentration range in a reproducible way with respect to gas and particulate phases, and also demonstrated reproducible particle deposition depending on smoke concentration. Furthermore, it was shown that the exposed cells maintained their viability. Native cigarette mainstream smoke induced dose-dependent cellular effects in exposed cells with respect to cellular viability (viable cell number monitored by tetrazolium salt cleavage) and intracellular parameters (ATP and glutathione content). Therefore, fresh, physically and chemically unmodified cigarette mainstream smoke can be tested using this novel system.
已有多种方法用于体外测试香烟烟雾的生物活性。但没有一种方法能够以实际的方式使培养的肺细胞暴露于构成香烟烟雾的复杂气态和颗粒混合物中。我们设计了一种能够实现这一目标的系统。本文介绍的系统能够使人肺细胞直接暴露于天然、未改性的香烟主流烟雾中。它由一台吸烟机、一个烟雾稀释装置、用于在线监测的分析装置以及一个基于Cultex**细胞培养系统的经过特殊改装的暴露模块组成，该暴露模块配备有气体暴露顶部。由于暴露装置的特殊设计和优化的暴露条件，该设备能够使培养的人肺细胞暴露于新鲜产生的香烟主流烟雾中。探索性实验表明，烟雾在气相和颗粒相方面能够在很宽的浓度范围内以可重复的方式进行稀释，并且还表明颗粒沉积具有可重复性，这取决于烟雾浓度。此外，结果显示暴露后的细胞保持了活力。天然香烟主流烟雾对暴露细胞的细胞活力（通过四唑盐裂解监测活细胞数量）和细胞内参数（ATP和谷胱甘肽含量）产生了剂量依赖性的细胞效应。因此，使用这个新系统可以测试新鲜的、物理和化学性质未改性的香烟主流烟雾。
