Institute of Neurophysiology and Center for Molecular Medicine Cologne (CMMC), University of Cologne (UKK), Robert-Koch-Str. 39, 50931, Cologne, Germany.
Department of In Vitro Toxicology and Dermato-Cosmetology, Faculty of Medicine and Pharmacy, Vrije Universiteit Brussel, Laarbeeklaan 103, 1090, Brussels, Belgium.
Arch Toxicol. 2018 Jan;92(1):371-381. doi: 10.1007/s00204-017-2065-z. Epub 2017 Sep 22.
There is a large demand of a human relevant in vitro test system suitable for assessing the cardiotoxic potential of cosmetic ingredients and other chemicals. Using human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), we have already established an in vitro cardiotoxicity assay and identified genomic biomarkers of anthracycline-induced cardiotoxicity in our previous work. Here, five cosmetic ingredients were studied by the new hiPSC-CMs test; kojic acid (KJA), triclosan (TS), triclocarban (TCC), 2,7-naphthalenediol (NPT), and basic red 51 (BR51) based on cytotoxicity as well as ATP assays, beating rate, and genomic biomarkers to determine the lowest observed effect concentration (LOEC) and no observed effect concentration (NOEC). The LOEC for beating rate were 400, 10, 3, >400, and 3 µM for KJA, TS, TCC, NPT, and BR51, respectively. The corresponding concentrations for cytotoxicity or ATP depletion were similar, with the exception of TS and TCC, where the cardiomyocyte-beating assay showed positive results at non-cytotoxic concentrations. Functional analysis also showed that the individual compounds caused different effects on hiPSC-CMs. While exposure to KJA, TS, TCC, and BR51 induced significant arrhythmic beating, NPT slightly decreased cell viability, but did not influence beating. Gene expression studies showed that TS and NPT caused down-regulation of cytoskeletal and cardiac ion homeostasis genes. Moreover, TS and NPT deregulated genomic biomarkers known to be affected also by anthracyclines. The present study demonstrates that hiPSC-CMs can be used to determine LOECs and NOECs in vitro, which can be compared to human blood concentrations to determine margins of exposure. Our in vitro assay, which so far has been tested with several anthracyclines and cosmetics, still requires validation by larger numbers of positive and negative controls, before it can be recommended for routine analysis.
人们对适合评估化妆品成分和其他化学物质心脏毒性的人类相关体外测试系统有很大的需求。在之前的工作中，我们使用人诱导多能干细胞衍生的心肌细胞（hiPSC-CMs）建立了一种体外心脏毒性测定法，并鉴定了蒽环类药物诱导心脏毒性的基因组生物标志物。在这里，我们使用新的 hiPSC-CMs 测试方法研究了五种化妆品成分；曲酸（KJA）、三氯生（TS）、三氯卡班（TCC）、2,7-萘二酚（NPT）和碱性红 51（BR51），通过细胞毒性和 ATP 测定、心率和基因组生物标志物来确定最低观察到的效应浓度（LOEC）和无观察到的效应浓度（NOEC）。KJA、TS、TCC、NPT 和 BR51 的心率 LOEC 分别为 400、10、3、>400 和 3μM。细胞毒性或 ATP 耗竭的相应浓度相似，除了 TS 和 TCC，其中在非细胞毒性浓度下，心肌细胞跳动测定显示出阳性结果。功能分析还表明，个别化合物对 hiPSC-CMs 产生不同的影响。虽然 KJA、TS、TCC 和 BR51 的暴露导致心律失常跳动显著增加，NPT 略微降低细胞活力，但不影响跳动。基因表达研究表明，TS 和 NPT 导致细胞骨架和心脏离子稳态基因的下调。此外，TS 和 NPT 使已知也受蒽环类药物影响的基因组生物标志物失调。本研究表明，hiPSC-CMs 可用于体外测定 LOEC 和 NOEC，可与人体血液浓度进行比较，以确定接触量。我们的体外测定法迄今为止已经用几种蒽环类药物和化妆品进行了测试，在推荐用于常规分析之前，仍然需要用更多的阳性和阴性对照进行验证。
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