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A novel in vitro exposure technique for toxicity testing of selected volatile organic compounds

机译:一种用于选择挥发性有机化合物毒性测试的新型体外暴露技术

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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-suffophenyl)-2H-tetrazolium) and NRU (neutral red uptake) assays following 24 h incubation. Airborne IC50 (50% inhibitory concentration) values were determined using dose response curves for xylene (IC50 = 5350 +/- 328 ppm, NRU; IC50 = 5750 +/- 433 ppm, NITS in skin fibroblast) and toluene (IC50 = 10 500 +/- 527 ppm, NRU; IC50 = 11200 +/- 1044 ppm, NITS 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.
机译:暴露于挥发性化学物质的蒸气是主要的职业和环境健康问题。由于挥发性有机化合物(VOC)的高挥发性和/或低溶解度,其毒性测试一直面临重大的技术问题。这项研究的目的是开发一种实用且可重现的体外暴露技术,用于VOC的毒性测试。使用静态方法在玻璃室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°C的空气/液体界面直接暴露于各种空气中浓度的选定VOCs 1小时。使用MTS(3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲氧基苯基)-2-(4-uffophenylphenyl)-2H-四唑鎓)和NRU(中性红吸收)研究了测试化学品的细胞毒性)孵育24小时后进行分析。使用二甲苯(IC50 = 5350 +/- 328 ppm,NRU; IC50 = 5750 +/- 433 ppm,皮肤成纤维细胞中的NITS)的剂量响应曲线确定空气传播的IC50(50%抑制浓度)值+/- 527 ppm,NRU; IC50 = 11200 +/- 1044 ppm,皮肤成纤维细胞中的NITS)。我们的研究结果表明,在气/液界面处直接静态暴露是用于VOC毒性测试的实用且可重现的技术。此外,该技术可用于体外挥发性化学物质的吸入和皮肤毒性研究,因为该方法密切模拟了体内的暴露方式。

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