title: Graphene and carbon nanotube nanocomposite for gene transfection.
abstract: Graphene and carbon nanotube nanocomposite (GCN) was synthesised and applied in gene transfection of pIRES plasmid conjugated with green fluorescent protein (GFP) in NIH-3T3 and NG97 cell lines. The tips of the multi-walled carbon nanotubes (MWCNTs) were exfoliated by oxygen plasma etching, which is also known to attach oxygen content groups on the MWCNT surfaces, changing their hydrophobicity. The nanocomposite was characterised by high resolution scanning electron microscopy; energy-dispersive X-ray, Fourier transform infrared and Raman spectroscopies, as well as zeta potential and particle size analyses using dynamic light scattering. BET adsorption isotherms showed the GCN to have an effective surface area of 38.5m(2)/g. The GCN and pIRES plasmid conjugated with the GFP gene, forming π-stacking when dispersed in water by magnetic stirring, resulting in a helical wrap. The measured zeta potential confirmed that the plasmid was connected to the nanocomposite. The NIH-3T3 and NG97 cell lines could phagocytize this wrap. The gene transfection was characterised by fluorescent protein produced in the cells and pictured by fluorescent microscopy. Before application, we studied GCN cell viability in NIH-3T3 and NG97 line cells using both MTT and Neutral Red uptake assays. Our results suggest that GCN has moderate stability behaviour as colloid solution and has great potential as a gene carrier agent in non-viral based therapy, with low cytotoxicity and good transfection efficiency.
authors.affiliation: Laboratory of Biotechnology, Department of Biochemistry, Institute of Biology at UNICAMP, Rua Monteiro Lobato 255, Campinas, SP CEP 13083-862, Brazil.
authors.forename: L M
authors.initials: LM
authors.lastname: Hollanda
authors.affiliation: Laboratory of Biomedical Nanotechnology, Institute of Research and Development at the UNIVAP, Av. Shishima Hifumi, 2911, CEP: 12244-000 Sao Jose dos Campos, SP, Brazil.
authors.forename: A O
authors.initials: AO
authors.lastname: Lobo
authors.affiliation: Laboratory of Biotechnology, Department of Biochemistry, Institute of Biology at UNICAMP, Rua Monteiro Lobato 255, Campinas, SP CEP 13083-862, Brazil.
authors.forename: M
authors.initials: M
authors.lastname: Lancellotti
authors.affiliation: Biological Chemistry Laboratory, Department of Physical Chemistry, Institute of Chemistry at UNICAMP, R. José de Castro, Campinas, SP CEP 13083-970, Brazil.
authors.forename: E
authors.initials: E
authors.lastname: Berni
authors.affiliation: Associated Laboratory of Sensors and Materials of the INPE, Av. dos Astronautas 1758, Sao Jose dos Campos CEP: 12227-010 SP, Brazil.
authors.forename: E J
authors.initials: EJ
authors.lastname: Corat
authors.affiliation: Associated Laboratory of Sensors and Materials of the INPE, Av. dos Astronautas 1758, Sao Jose dos Campos CEP: 12227-010 SP, Brazil. Electronic address: hudsonzanin@gmail.com.
authors.forename: H
authors.initials: H
authors.lastname: Zanin
mesh_terms: D016475:3T3 Cells; D000327:Adsorption; D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival; D006108:Graphite / Q000737:chemistry*; D049452:Green Fluorescent Proteins; D006801:Humans; D008280:Magnetics; D051379:Mice; D008855:Microscopy, Electron, Scanning; D053761:Nanocomposites / Q000737:chemistry*; D037742:Nanotubes, Carbon / Q000737:chemistry*; D014162:Transfection; D014867:Water
chemical_list: D037742:Nanotubes, Carbon; D014867:Water; D049452:Green Fluorescent Proteins; D006108:Graphite
keywords: Carbon nanotubes; Cell viability; Composite; Gene transfer; Graphene oxide; Plasma