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J Biomed Mater Res A. 2018 Jun;106(6):1500-1510. doi: 10.1002/jbm.a.36354. Epub 2018 Feb 13.

Carboxylated graphene oxide promoted axonal guidance growth by activating Netrin-1/deleted in colorectal cancer signaling in rat primary cultured cortical neurons.

Liu M1,2, Jia Z1,2, Xiao X1,2, Zhang Z1,2, Li P1,2, Zhou G1,2, Fan Y1,2,3.

Author information

1
Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical Engineering, Beihang University, Beijing, 100191, China.
2
Beijing Advanced Innovation Centre for Biomedical Engineering, Beihang University, Beijing, 102402, China.
3
National Research Center for Rehabilitation Technical Aids, Beijing, 100176, China.

Abstract

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.

KEYWORDS:

Netrin-1; axonal guidance growth; carboxylated graphene oxide; graphene oxide; nerve regeneration

PMID:
29377591
DOI:
10.1002/jbm.a.36354

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