Graphene oxide (GO) and reduced graphene oxide (rGO) are carbon-based nanomaterials that have a wide range of applicability. Therefore, it is expected that their residual traces reach the aquatic environment, accumulate, and interact with its different compartments and the biota living in them. The concentration- and time-dependency response to GO and rGO in aquatic organisms are still poorly known. In the present study, the effects of GO and rGO on zebrafish hepatocytes were investigated using in vitro assays performed with established liver cell lines from zebrafish (ZFL). GO and rGO nanosheets were applied on ZFL cells at a concentration range of 1-100 µg mL⁻¹ for 24 and 72 h. The internalization of GO and rGO nanosheets, reactive oxygen species (ROS) production, cell viability, and cell death were evaluated. The internalization of GO increased as the concentrations of GO increased. The rGO nanosheets were smaller than GO nanosheets, and their hydrophobic characteristic favors their interaction with the cell membrane. However, the rGO nanosheets were not observed in the uptake assay. Exposure for 72 h was found to cause harmful effects in ZFL cells, causing higher ROS production in cells exposed to rGO and stopping cell replication. Nevertheless, GO did not stop cell replication, but exposed cells had higher levels of apoptosis and necrosis. After 72 h, both GO and rGO were toxic, but with different mechanisms of toxicity.

氧化石墨烯（GO）和还原氧化石墨烯（rGO）是具有广泛适用性的碳基纳米材料。因此，预计它们的残留痕迹会到达水生环境，积累并与其不同的隔间和生活在其中的生物群相互作用。对水生生物中 GO 和 rGO 的浓度和时间依赖性响应仍然知之甚少。在本研究中，使用已建立的斑马鱼肝细胞系 (ZFL) 进行的体外测定研究了 GO 和 rGO 对斑马鱼肝细胞的影响。^{GO 和 rGO 纳米片以 1-100 µg mL -1}的浓度范围应用于 ZFL 细胞24 和 72 小时。评估了 GO 和 rGO 纳米片的内化、活性氧 (ROS) 的产生、细胞活力和细胞死亡。GO 的内化随着 GO 浓度的增加而增加。rGO 纳米片比 GO 纳米片小，它们的疏水特性有利于它们与细胞膜的相互作用。然而，在吸收测定中没有观察到 rGO 纳米片。发现暴露 72 小时会对 ZFL 细胞造成有害影响，导致暴露于 rGO 的细胞产生更高的 ROS 并阻止细胞复制。尽管如此，GO 并没有停止细胞复制，但暴露的细胞具有更高水平的细胞凋亡和坏死。72 小时后，GO 和 rGO 均具有毒性，但毒性机制不同。