title: Carbon nanoparticles for gene transfection in eukaryotic cell lines.
abstract: For the first time, oxygen terminated cellulose carbon nanoparticles (CCN) was synthesised and applied in gene transfection of pIRES plasmid. The CCN was prepared from catalytic of polyaniline by chemical vapour deposition techniques. This plasmid contains one gene that encodes the green fluorescent protein (GFP) in eukaryotic cells, making them fluorescent. This new nanomaterial and pIRES plasmid formed π-stacking when dispersed in water by magnetic stirring. The frequencies shift in zeta potential confirmed the plasmid strongly connects to the nanomaterial. In vitro tests found that this conjugation was phagocytised by NG97, NIH-3T3 and A549 cell lines making them fluorescent, which was visualised by fluorescent microscopy. Before the transfection test, we studied CCN in cell viability. Both MTT and Neutral Red uptake tests were carried out using NG97, NIH-3T3 and A549 cell lines. Further, we use metabolomics to verify if small amounts of nanomaterial would be enough to cause some cellular damage in NG97 cells. We showed two mechanisms of action by CCN-DNA complex, producing an exogenous protein by the transfected cell and metabolomic changes that contributed by better understanding of glioblastoma, being the major finding of this work. Our results suggested that this nanomaterial has great potential as a gene carrier agent in non-viral based therapy, with low cytotoxicity, good transfection efficiency, and low cell damage in small amounts of nanomaterials in metabolomic tests.
authors.affiliation: Departamento de Semicondutores, Instrumentos e Fotônica, Faculdade de Engenharia Elétrica e de Computação, Universidade Estadual de Campinas, 13083-852 Campinas, SP, Brazil. Electronic address: hudsonzanin@gmail.com.
authors.forename: H
authors.initials: H
authors.lastname: Zanin
authors.affiliation: Laboratory of Biotechnology, Department of Biochemistry, Institute of Biology at UNICAMP, Rua Monteiro Lobato 255, Campinas, SP CEP 13083-862, Brazil. Electronic address: luciana.hollanda@gmail.com.
authors.forename: L M
authors.initials: LM
authors.lastname: Hollanda
authors.affiliation: Departamento de Semicondutores, Instrumentos e Fotônica, Faculdade de Engenharia Elétrica e de Computação, Universidade Estadual de Campinas, 13083-852 Campinas, SP, Brazil.
authors.forename: H J
authors.initials: HJ
authors.lastname: Ceragioli
authors.affiliation: Innovare Biomarkers Laboratory, Medicine and Experimental Surgery Nucleus, UNICAMP, Rua Cinco de Junho, 350, Campinas, São Paulo CEP 13083-877, Brazil.
authors.forename: M S
authors.initials: MS
authors.lastname: Ferreira
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: D
authors.initials: D
authors.lastname: Machado
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: Innovare Biomarkers Laboratory, Medicine and Experimental Surgery Nucleus, UNICAMP, Rua Cinco de Junho, 350, Campinas, São Paulo CEP 13083-877, Brazil.
authors.forename: R R
authors.initials: RR
authors.lastname: Catharino
authors.affiliation: Departamento de Semicondutores, Instrumentos e Fotônica, Faculdade de Engenharia Elétrica e de Computação, Universidade Estadual de Campinas, 13083-852 Campinas, SP, Brazil.
authors.forename: V
authors.initials: V
authors.lastname: Baranauskas
authors.affiliation: Laboratory of Biomedical Nanotechnology (NANOBIO), Universidade do Vale do Paraiba (UNIVAP), Av. Shishima Hifumi 2911, Sao Jose dos Campos 12224-000, SP, Brazil.
authors.forename: A O
authors.initials: AO
authors.lastname: Lobo
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002244:Carbon / Q000737:chemistry*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002482:Cellulose / Q000031:analogs & derivatives / Q000737:chemistry; D004247:DNA / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D055432:Metabolomics / Q000379:methods; D051379:Mice; D008855:Microscopy, Electron, Scanning; D053758:Nanoparticles / Q000737:chemistry*; D010316:Particle Size; D010957:Plasmids / Q000235:genetics; D017550:Spectroscopy, Fourier Transform Infrared; D013059:Spectrum Analysis, Raman; D014162:Transfection
chemical_list: C033190:DNA-cellulose; D049452:Green Fluorescent Proteins; D002244:Carbon; D002482:Cellulose; D004247:DNA
keywords: Carbon nanomaterial; Cell viability; Gene transfection; Metabolomic test