Production of 2-D nanocarbons with different bioaffinities as ROS quenching agents to improve mammalian oocyte cryopreservation outcomes

我们根据之前的方法（Hummer的方法）制备氧化石墨烯（GO），并在80°C下缓慢进行热还原长达2周，以确保官能团的成功还原得到还原氧化石墨烯（rGO）。采用FTIR、TGA、XPS和XRD对氧化石墨烯和还原氧化石墨烯样品进行了结构分析。采用ESR法分析其活性氧（ROS）活性。此外，为了了解它们在培养基设置中使用的能力，将样品提交给分散测试。根据所有测试结果，选择原始氧化石墨烯（氧化水平30-35%）在非酶促抗氧化剂存在下分析其在卵母细胞体外成熟（IVM）过程中的毒性。结果表明，在卵母细胞IVM过程中添加氧化石墨烯对卵母细胞活力没有显著的负面影响。

We prepared graphene oxide (GO) according to a previous method (Hummer’s method) and slowly performed thermal reduction at 80°C up to 2 weeks to ensure a successful reduction of the functional groups to obtain reduced graphene oxide (rGO). The GO and rGO samples were structurally analyzed by FTIR, TGA, XPS and XRD. To analyze their reactive oxygen species (ROS) activity, they were analyzed by ESR. Furthermore, to understand their ability to be used in a culture medium setting, the samples were submitted to a dispersion test.According to the results of all the tests, pristine GO (oxidation level 30-35%) was selected to analyze its toxicity during oocyte in vitro maturation (IVM) in the presence of non-enzymatic antioxidants. Results suggest that the addition of GO during oocyte IVM does not negatively affect oocyte viability significantly.