Garry V F, Opp C W, Wiencke J K, Lakatua D
Pharmacology. 1982;25(4):214-21. doi: 10.1159/000137745.
In an earlier study, we demonstrated that human exposure to ethylene oxide (EO) during sterilization procedures was associated with increased sister chromatid exchanges (SCE) in cultured peripheral blood. To study further toxic and mutagenic properties of EO in vitro, we devised a membrane dosimetry system which can be used to expose lymphocytes in peripheral blood to specified amounts of EO. The system consists of an exposure chamber, cell culture dishes to which are applied plastic membranes of varying diameters and an infrared analyzer used to monitor ambient EO concentrations in the system. Aliquots of EO-exposed media were subjected to gas chromatography for quantitation. Our preliminary analyses of the cultured cells indicate that the membrane dosimetry system we developed is capable of reproducibly delivering quantities of EO to target cells. Elevated SCEs were observed at as little as 10 micrograms/ml (in media) during a 20-min exposure period. A significant dose-response relationship between SCE and EO dose was recorded up to 35 micrograms/ml, the highest dose tested. Lymphocytes at 35 micrograms/ml showed a fourfold increase in SCE compared to control cultures. We conclude that the membrane dosimetry system provides a reproducible method to investigate EO mutagenesis in human cells in vitro and may prove applicable to a wide range of mutagenic gases.
在早期的一项研究中,我们证明了人类在灭菌过程中接触环氧乙烷(EO)与培养的外周血中姐妹染色单体交换(SCE)增加有关。为了进一步研究EO在体外的毒性和致突变特性,我们设计了一种膜剂量测定系统,该系统可用于使外周血中的淋巴细胞接触特定量的EO。该系统由一个暴露室、应用不同直径塑料膜的细胞培养皿以及用于监测系统中环境EO浓度的红外分析仪组成。将暴露于EO的培养基等分试样进行气相色谱定量分析。我们对培养细胞的初步分析表明,我们开发的膜剂量测定系统能够可重复地将一定量的EO输送到靶细胞。在20分钟的暴露期内,在低至10微克/毫升(培养基中)时就观察到SCE升高。在高达35微克/毫升(测试的最高剂量)时,记录到SCE与EO剂量之间存在显著的剂量反应关系。与对照培养物相比,35微克/毫升的淋巴细胞SCE增加了四倍。我们得出结论,膜剂量测定系统提供了一种可重复的方法来研究体外人类细胞中的EO诱变作用,并且可能证明适用于广泛的诱变气体。