Carboxylated graphene oxide promoted axonal guidance growth by activating Netrin-1/deleted in colorectal cancer signaling in rat primary cultured cortical neurons
Carboxylated graphene oxide promoted axonal guidance growth by activating Netrin-1/deleted in colorectal cancer signaling in rat primary cultured cortical neurons
复制标题
羧化氧化石墨烯通过激活大鼠原代培养皮层神经元结直肠癌信号传导中的 Netrin-1/缺失促进轴突引导生长
DOI：
10.1002/jbm.a.36354
复制
发表时间：
2018
影响因子：
4.9
通讯作者：
Fan Yubo
中科院分区：
文献类型：
--
作者：
Liu Meili;Jia Zhengtai;Xiao Xiongfu;Zhang Zhifa;Li Ping;Zhou Gang;Fan Yubo
石墨烯及其衍生物纳米材料近年来得到了广泛的应用，但其对神经元引导生长的影响尚未见报道。在本研究中，氧化石墨烯（GO）和羧化氧化石墨烯（GO-COOH）被用来研究在原代培养的皮层神经元轴突导向生长的潜在影响。此外，我们使用傅里叶变换红外分光光度计和扫描电子显微镜分析以及拉曼分析来表征合成的GO和GO-COOH的结构和化学组成。GO不具有神经毒性，并且在可溶形式下不导电。然而，GO-COOH具有更高的溶解度和导电性。在GO和GO-COOH处理（0、1、2、4、5、6、8、10、12、14、16、18、20、50和100 µg/mL）后，使用CCK-8测定和荧光素二乙酸酯评估细胞活力。在GO（2和4 μg/mL）和GO-COOH处理（2和4 μ g/mL）后，细胞活力和轴突生长显著增加。我们进一步研究了GO和GO-COOH（2和4 μg/mL）应用后轴突导向生长的分子机制。此外，GO和GO-COOH上调Netrin-1及其受体的表达，通过免疫荧光测定和蛋白质印迹测定在结直肠癌中缺失。我们的研究表明，GO-COOH激活Cdc 42和Rac 1，并显着降低RhoA。因此，在本研究中，GO-COOH（2 µg/mL）作为轴突引导和生长的纳米载体比GO好得多。GO-COOH可用于促进未来神经元再生的指导。© 2018 Wiley Periodicals，Inc. J Biomed Mater Res Part A：106A：1500-1510，2018.
Nanomaterials of graphene and its derivatives have been widely applied in recent years, but whose impacts on the neuronal guidance growth are still not reported. In the present study, graphene oxide (GO) and carboxylated graphene oxide (GO‐COOH) were used to investigate the potential effects on axonal guidance growth in the primary cultured cortical neurons. In addition, we characterized the structure and chemical composition of synthesized GO and GO‐COOH using Fourier transform infrared spectrophotometer and scanning electron microscope assays and Raman analysis. GO is not neurotoxic and not conductive in a soluble form. However, GO‐COOH has higher solubility and conductivity. Cell viability was assessed using CCK‐8 assays and fluorescein diacetate after GO and GO‐COOH treatment (0, 1, 2, 4, 5, 6, 8, 10, 12, 14, 16, 18, 20, 50, and 100 µg/mL). There are significant increases of cell viability and axonal growth after GO (2 and 4 μg/mL) and GO‐COOH treatment (2 and 4 μg/mL). We further investigated the molecular mechanism of axonal guidance growth after GO and GO‐COOH (2 and 4 μg/mL) application. Additionally, GO and GO‐COOH up‐regulated expression of Netrin‐1 and its receptor, deleted in colorectal cancer by immunofluorescence assays and western blots assay. Our study demonstrated that GO‐COOH activated Cdc42 and Rac1 and dramatically decreased RhoA. Thus, GO‐COOH (2 µg/mL) is much better to be nanocarriers than GO for axonal guidance and growth in this study. GO‐COOH may be used to facilitate guidance for regenerating neurons in the future. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1500–1510, 2018.