Bakand S, Winder C, Khalil C, Hayes A
Chemical Safety and Applied Toxicology (CSAT) Laboratories, School of Safety Science, The University of New South Wales, UNSW, Sydney, 2052, Australia.
J Environ Monit. 2006 Jan;8(1):100-5. doi: 10.1039/b509812b. Epub 2005 Nov 9.
Exposure to vapours of volatile chemicals is a major occupational and environmental health concern. Toxicity testing of volatile organic compounds (VOCs) has always faced significant technological problems due to their high volatility and/or low solubility. The aim of this study was to develop a practical and reproducible in vitro exposure technique for toxicity testing of VOCs. Standard test atmospheres of xylene and toluene were generated in glass chambers using a static method. Human cells including: A549-lung derived cell lines, HepG2-liver derived cell lines and skin fibroblasts, were grown in porous membranes and exposed to various airborne concentrations of selected VOCs directly at the air/liquid interface for 1 h at 37 degrees C. Cytotoxicity of test chemicals was investigated using the MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) and NRU (neutral red uptake) assays following 24 h incubation. Airborne IC(50) (50% inhibitory concentration) values were determined using dose response curves for xylene (IC(50)=5350+/- 328 ppm, NRU; IC(50)=5750+/- 433 ppm, MTS in skin fibroblast) and toluene (IC(50)=0 500+/- 527 ppm, NRU; IC(50)=11,200 +/- 1,044 ppm, MTS in skin fibroblast). Our findings suggest that static direct exposure at the air/liquid interface is a practical and reproducible technique for toxicity testing of VOCs. Further, this technique can be used for inhalational and dermal toxicity studies of volatile chemicals in vitro as the exposure pattern in vivo is closely simulated by this method.
接触挥发性化学物质的蒸气是一个主要的职业和环境健康问题。由于挥发性有机化合物(VOCs)具有高挥发性和/或低溶解性,其毒性测试一直面临重大技术难题。本研究的目的是开发一种实用且可重复的体外暴露技术,用于VOCs的毒性测试。使用静态方法在玻璃 chamber 中生成二甲苯和甲苯的标准测试气氛。包括A549肺源性细胞系、HepG2肝源性细胞系和皮肤成纤维细胞在内的人类细胞在多孔膜中生长,并在37℃下于气/液界面直接暴露于选定VOCs的各种空气浓度下1小时。在孵育24小时后,使用MTS(3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲氧基苯基)-2-(4-磺基苯基)-2H-四唑)和NRU(中性红摄取)测定法研究测试化学品的细胞毒性。使用二甲苯(IC(50)=5350±328 ppm,NRU;IC(50)=5750±433 ppm,皮肤成纤维细胞中的MTS)和甲苯(IC(50)=0 500±527 ppm,NRU;IC(50)=11,200±1,044 ppm,皮肤成纤维细胞中的MTS)的剂量反应曲线确定空气IC(50)(50%抑制浓度)值。我们的研究结果表明,气/液界面的静态直接暴露是一种实用且可重复的VOCs毒性测试技术。此外,由于该方法紧密模拟了体内暴露模式,该技术可用于挥发性化学物质的体外吸入和皮肤毒性研究。