Michel Christian, Herzog Simon, de Capitani Christian, Burkhardt-Holm Patricia, Pietsch Constanze
Man-Society-Environment (Programm MGU), Department of Environmental Sciences, University of Basel, Basel, Switzerland.
Mineralogy-Petrography, Department of Environmental Sciences, University of Basel, Basel, Switzerland.
PLoS One. 2014 Jul 3;9(7):e100856. doi: 10.1371/journal.pone.0100856. eCollection 2014.
Worldwide increases in fluvial fine sediment are a threat to aquatic animal health. Fluvial fine sediment is always a mixture of particles whose mineralogical composition differs depending on the sediment source and catchment area geology. Nonetheless, whether particle impact in aquatic organisms differs between mineral species remains to be investigated. This study applied an in vitro approach to evaluate cytotoxicity and uptake of four common fluvial mineral particles (quartz, feldspar, mica, and kaolin; concentrations: 10, 50, 250 mg L(-1)) in the rainbow trout epithelial gill cell line RTgill-W1. Cells were exposed for 24, 48, 72, and 96 h. Cytotoxicity assays for cell membrane integrity (propidium iodide assay), oxidative stress (H2DCF-DA assay), and metabolic activity (MTT assay) were applied. These assays were complemented with cell counts and transmission electron microscopy. Regardless of mineral species, particles ≤ 2 µm in diameter were taken up by the cells, suggesting that particles of all mineral species came into contact and interacted with the cells. Not all particles, however, caused strong cytotoxicity: Among all assays the tectosilicates quartz and feldspar caused sporadic maximum changes of 0.8-1.2-fold compared to controls. In contrast, cytotoxicity of the clay particles was distinctly stronger and even differed between the two particle types: mica induced concentration-dependent increases in free radicals, with consistent 1.6-1.8-fold-changes at the 250 mg L(-1) concentration, and a dilated endoplasmic reticulum. Kaolin caused concentration-dependent increases in cell membrane damage, with consistent 1.3-1.6-fold increases at the 250 mg L(-1) concentration. All effects occurred in the presence or absence of 10% fetal bovine serum. Cell numbers per se were marginally affected. Results indicate that (i.) natural mineral particles can be cytotoxic to gill epithelial cells, (ii.) their cytotoxic potential differs between mineral species, with clay particles being more cytotoxic, and (iii.) some clays might induce effects comparable to engineered nanoparticles.
全球范围内河流细颗粒沉积物的增加对水生动物健康构成威胁。河流细颗粒沉积物总是由颗粒混合而成，其矿物成分因沉积物来源和集水区地质情况而异。然而，不同矿物种类的颗粒对水生生物的影响是否存在差异仍有待研究。本研究采用体外方法评估四种常见河流矿物颗粒（石英、长石、云母和高岭土；浓度：10、50、250 mg L(-1)）在虹鳟鱼鳃上皮细胞系RTgill-W1中的细胞毒性和摄取情况。细胞暴露24、48、72和96小时。应用了针对细胞膜完整性（碘化丙啶检测）、氧化应激（H2DCF-DA检测）和代谢活性（MTT检测）的细胞毒性检测方法。这些检测方法辅以细胞计数和透射电子显微镜观察。无论矿物种类如何，直径≤2 µm的颗粒都会被细胞摄取，这表明所有矿物种类的颗粒都与细胞发生了接触并相互作用。然而，并非所有颗粒都会引起强烈的细胞毒性：在所有检测中，架状硅酸盐石英和长石与对照组相比，偶尔会出现0.8 - 1.2倍的最大变化。相比之下，粘土颗粒的细胞毒性明显更强，甚至在两种颗粒类型之间也存在差异：云母诱导自由基浓度依赖性增加，在250 mg L(-1)浓度下一致出现1.6 - 1.8倍的变化，并且内质网扩张。高岭土导致细胞膜损伤浓度依赖性增加，在250 mg L(-1)浓度下一致出现1.3 - 1.6倍的增加。所有这些效应在有或没有10%胎牛血清的情况下都会发生。细胞数量本身受到的影响很小。结果表明：（i.）天然矿物颗粒可能对鳃上皮细胞具有细胞毒性，（ii.）它们的细胞毒性潜力因矿物种类而异，粘土颗粒的细胞毒性更强，（iii.）一些粘土可能会诱导与工程纳米颗粒相当的效应。
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