Horn Gabriele, Schmitt Christian, Kehe Kai, Siegert Markus, Worek Franz, Amend Niko
Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstrasse 11, 80937, Munich, Germany.
Faculty of Medicine, Walther-Straub-Institute of Pharmacology and Toxicology, Ludwig-Maximilians-Universität München, Goethestr. 33, 80336, Munich, Germany.
Arch Toxicol. 2025 Apr 22. doi: 10.1007/s00204-025-04059-3.
Cell cultivation tools are usually not considered the primary factors influencing research results. In this study, we selected two organophosphorus compounds (OP) to examine possible compound absorption in five different commercially available cell culture plates. In addition, compound degradation processes were studied. The plates were exposed to the nerve agent VX or the OP pesticide parathion. Using analytical methods such as LC-ESI-MS/MS or GC-MS, the respective concentrations of VX and its degradation products O-ethyl methylphosphonic acid (EMPA) and S-[2-(diisopropylamino)ethyl]methylphosphonothioic acid (EA-2192) as well as parathion, paraoxon, diethyl thiophosphate (DETP), and diethyl phosphate (DEP) were determined up to 72 h. Our data showed that the choice of the cell culture plate can have an impact on the OP concentration and the degradation products. The balance of VX, EMPA, and EA-2192 resulted in about the initial VX concentration at all evaluated time points, indicating that VX absorption was not likely to occur in the cell culture plates. In contrast, relevant parathion concentrations were unaccounted for after 72 h suggesting absorption to the plates. In conclusion, it was shown for the model substance parathion that it is important to characterize cell culture tools, i.e., the respective plates, regarding possible compound interaction. The suitability of the cell cultivation tool for investigations with an OP should be evaluated in advance to obtain a high quality of the study and to improve the concordance with in vivo data.
细胞培养工具通常不被视为影响研究结果的主要因素。在本研究中，我们选择了两种有机磷化合物（OP），以检测其在五种不同市售细胞培养板中的可能吸收情况。此外，还研究了化合物的降解过程。将这些培养板暴露于神经毒剂VX或OP农药对硫磷中。使用诸如液相色谱-电喷雾串联质谱法（LC-ESI-MS/MS）或气相色谱-质谱法（GC-MS）等分析方法，测定了VX及其降解产物O-乙基甲基膦酸（EMPA）和S-[2-(二异丙基氨基)乙基]甲基硫代膦酸（EA-2192）以及对硫磷、对氧磷、二乙基硫代磷酸酯（DETP）和二乙基磷酸酯（DEP）在72小时内的各自浓度。我们的数据表明，细胞培养板的选择可能会对OP浓度和降解产物产生影响。在所有评估时间点，VX、EMPA和EA-2192的平衡导致约为初始VX浓度，这表明在细胞培养板中不太可能发生VX吸收。相比之下，72小时后对硫磷的相关浓度未得到解释，这表明其被培养板吸收。总之，对于模型物质对硫磷而言，表征细胞培养工具（即各自的培养板）关于可能的化合物相互作用是很重要的。应提前评估细胞培养工具对于使用OP进行研究的适用性，以获得高质量的研究并提高与体内数据的一致性。
