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首页> 外文期刊>Environmental toxicology and chemistry >DEVELOPMENT OF A POLYDIMETHYLSILOXANE FILM-BASED PASSIVE DOSING METHOD IN THE IN VITRO DR-CALUX~® ASSAY
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DEVELOPMENT OF A POLYDIMETHYLSILOXANE FILM-BASED PASSIVE DOSING METHOD IN THE IN VITRO DR-CALUX~® ASSAY

机译:DR-CALUX〜®体外测定中基于聚二甲基硅氧烷薄膜的被动定量方法的开发

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摘要

In bioassays, exposure concentrations of test compounds are usually expressed as nominal concentrations. As a result of various processes, such as adsorption, degradation, or uptake, the actual freely dissolved concentration of the test compound may differ from the nominal concentration. The goal of the present study was to develop a method to dose passively the freely dissolved fraction of organic chemicals in an in vitro bioassay with adherent cells. To this end, a polydimethylsiloxane (PDMS) film-based method was developed for a reporter gene assay for dioxin-like compounds in a rat liver cell line. Polydimethylsiloxane films loaded with test compounds ensure that the concentration during exposure is in equilibrium and that the ratio between the concentration on the film and the concentration in medium is constant. Benzo[k]fluoranthene (BkF) was used as a model compound to develop the passive dosing method in transwell plates, which was further tested with a complex mixture, i.e., an extract prepared from a contaminated sediment. A higher dioxin-like activity was found when extracts were dosed by passive dosing with PDMS than when directly added to medium. Comparison with analysis of the concentration of BkF in medium shows that passive dosing of individual chemicals may not be necessary if freely dissolved concentrations are known. Use of PDMS for passive dosing of complex samples may represent a more realistic method for exposure in in vitro bioassays.
机译:在生物测定中,测试化合物的暴露浓度通常表示为标称浓度。由于各种过程(例如吸附,降解或吸收)的影响,测试化合物的实际自由溶解浓度​​可能与标称浓度不同。本研究的目的是开发一种在具有贴壁细胞的体外生物测定中被动剂量有机化学物质自由溶解部分的剂量的方法。为此,开发了一种基于聚二甲基硅氧烷(PDMS)膜的方法,用于大鼠肝细胞系中二恶英样化合物的报告基因测定。装有测试化合物的聚二甲基硅氧烷薄膜可确保曝光期间的浓度平衡,并且薄膜上的浓度与介质中的浓度之比恒定。苯并[k]荧蒽(BkF)被用作模型化合物,以开发多孔板中的被动加药方法,并用复杂的混合物(即从受污染的沉积物制备​​的提取物)进一步测试。与PDMS直接添加相比,通过PDMS被动给药提取物时,发现类二恶英活性更高。与培养基中BkF浓度分析的比较表明,如果已知可自由溶解的浓度,则无需对单个化学品进行被动加药。使用PDMS进行复杂样品的被动定量给药可能代表一种更现实的体外生物测定方法。

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