title: An in vitro evaluation of graphene oxide reduced by Ganoderma spp. in human breast cancer cells (MDA-MB-231).
abstract: BACKGROUND
  Recently, graphene and graphene-related materials have attracted much attention due their unique properties, such as their physical, chemical, and biocompatibility properties. This study aimed to determine the cytotoxic effects of graphene oxide (GO) that is reduced biologically using Ganoderma spp. mushroom extracts in MDA-MB-231 human breast cancer cells.
  
  
  METHODS
  Herein, we describe a facile and green method for the reduction of GO using extracts of Ganoderma spp. as a reducing agent. GO was reduced without any hazardous chemicals in an aqueous solution, and the reduced GO was characterized using a range of analytical procedures. The Ganoderma extract (GE)-reduced GO (GE-rGO) was characterized by ultraviolet-visible absorption spectroscopy, X-ray diffraction, Fourier-transform infrared spectroscopy, X-ray photoelectron spectroscopy, dynamic light scattering, scanning electron microscopy, Raman spectroscopy, and atomic force microscopy. Furthermore, the toxicity of GE-rGO was evaluated using a sequence of assays such as cell viability, lactate dehydrogenase leakage, and reactive oxygen species generation in human breast cancer cells (MDA-MB-231).
  
  
  RESULTS
  The preliminary characterization of reduction of GO was confirmed by the red-shifting of the absorption peak for GE-rGO to 265 nm from 230 nm. The size of GO and GE-rGO was found to be 1,880 and 3,200 nm, respectively. X-ray diffraction results confirmed that reduction processes of GO and the processes of removing intercalated water molecules and the oxide groups. The surface functionalities and chemical natures of GO and GE-rGO were confirmed using Fourier-transform infrared spectroscopy and X-ray photoelectron spectroscopy. The surface morphologies of the synthesized graphene were analyzed using high-resolution scanning electron microscopy. Raman spectroscopy revealed single- and multilayer properties of GE-rGO. Atomic force microscopy images provided evidence for the formation of graphene. Furthermore, the effect of GO and GE-rGO was examined using a series of assays, such as cell viability, membrane integrity, and reactive oxygen species generation, which are key molecules involved in apoptosis. The results obtained from cell viability and lactate dehydrogenase assay suggest that GO and GE-rGO cause dose-dependent toxicity in the cells. Interestingly, it was found that biologically derived GE-rGO is more toxic to cancer cells than GO.
  
  
  CONCLUSIONS
  We describe a simple, green, nontoxic, and cost-effective approach to producing graphene using mushroom extract as a reducing and stabilizing agent. The proposed method could enable synthesis of graphene with potential biological and biomedical applications such as in cancer and angiogenic disorders. To our knowledge, this is the first report using mushroom extract as a reducing agent for the synthesis of graphene. Mushroom extract can be used as a biocatalyst for the production of graphene.
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Seoul, South Korea ; GS Institute of Bio and Nanotechnology, Coimbatore, Tamilnadu, India.
authors.forename: Sangiliyandi
authors.initials: S
authors.lastname: Gurunathan
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Seoul, South Korea.
authors.forename: Jaewoong
authors.initials: J
authors.lastname: Han
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Seoul, South Korea.
authors.forename: Jung Hyun
authors.initials: JH
authors.lastname: Park
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Seoul, South Korea.
authors.forename: Jin Hoi
authors.initials: JH
authors.lastname: Kim
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000473:pathology; D002457:Cell Extracts / Q000737:chemistry*; D002458:Cell Fractionation; D045744:Cell Line, Tumor; D032902:Ganoderma / Q000737:chemistry*; D006108:Graphite / Q000008:administration & dosage* / Q000138:chemical synthesis*; D006801:Humans; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry*; D010084:Oxidation-Reduction; D010087:Oxides / Q000008:administration & dosage / Q000737:chemistry
chemical_list: D002457:Cell Extracts; D010087:Oxides; D006108:Graphite
keywords: Raman spectroscopy; UV-visible spectroscopy; X-ray diffraction; reduced graphene oxide
references.citation: J Mater Chem B. 2013 Sep 14;1(34):4267-4272
references.pmid: 32261022
references.citation: J Am Chem Soc. 2011 Jun 22;133(24):9148-51
references.pmid: 21604712
references.citation: J Am Chem Soc. 2009 Aug 12;131(31):11027-32
references.pmid: 19601624
references.citation: ACS Nano. 2010 Aug 24;4(8):4852-6
references.pmid: 20731460
references.citation: Phys Rev Lett. 2006 Nov 3;97(18):187401
references.pmid: 17155573
references.citation: ACS Nano. 2010 Apr 27;4(4):2429-37
references.pmid: 20359169
references.citation: Colloids Surf B Biointerfaces. 2013 Feb 1;102:772-7
references.pmid: 23107955
references.citation: Nano Lett. 2010 Oct 13;10(10):4061-6
references.pmid: 20738114
references.citation: Science. 2004 Oct 22;306(5696):666-9
references.pmid: 15499015
references.citation: ACS Nano. 2010 Jun 22;4(6):3181-6
references.pmid: 20481456
references.citation: ACS Nano. 2011 Sep 27;5(9):6971-80
references.pmid: 21851105
references.citation: ACS Nano. 2010 Oct 26;4(10):5731-6
references.pmid: 20925398
references.citation: J Colloid Interface Sci. 2012 Oct 1;383(1):36-42
references.pmid: 22795947
references.citation: ACS Nano. 2012 Mar 27;6(3):2731-40
references.pmid: 22376049
references.citation: Nano Lett. 2011 Dec 14;11(12):5201-7
references.pmid: 22023654
references.citation: Nano Lett. 2008 Dec;8(12):4469-76
references.pmid: 19367973
references.citation: Nanoscale Res Lett. 2011 Dec;6(1):8
references.pmid: 27502632
references.citation: Science. 2006 Feb 3;311(5761):622-7
references.pmid: 16456071
references.citation: Anal Chem. 2009 Jul 15;81(14):5603-13
references.pmid: 19522529
references.citation: Small. 2008 Jun;4(6):721-7
references.pmid: 18504717
references.citation: Nanotechnology. 2010 Apr 30;21(17):175602
references.pmid: 20357411
references.citation: Adv Mater. 2012 Jun 19;24(23):3134-7
references.pmid: 22570307
references.citation: Cancer Lett. 2004 Dec 8;216(1):9-20
references.pmid: 15500944
references.citation: Chemistry. 2002 Mar 15;8(6):1424-9
references.pmid: 11921226
references.citation: Int J Nanomedicine. 2013;8:1015-27
references.pmid: 23687445
references.citation: Nat Mater. 2007 Mar;6(3):183-91
references.pmid: 17330084
references.citation: Colloids Surf B Biointerfaces. 2013 Nov 1;111:376-83
references.pmid: 23850746
references.citation: Small. 2012 Feb 6;8(3):423-31
references.pmid: 22162356
references.citation: Colloids Surf B Biointerfaces. 2009 Jun 1;71(1):133-7
references.pmid: 19269142
references.citation: ACS Appl Mater Interfaces. 2011 Jul;3(7):2607-15
references.pmid: 21650218
references.citation: Colloids Surf B Biointerfaces. 2006 Feb 1;47(2):160-4
references.pmid: 16420977
references.citation: Chem Soc Rev. 2010 Jan;39(1):228-40
references.pmid: 20023850
references.citation: Biomaterials. 2012 Nov;33(32):8017-25
references.pmid: 22863381
references.citation: Biomaterials. 2012 Jan;33(2):402-11
references.pmid: 22019121
references.citation: Chem Commun (Camb). 2010 Feb 21;46(7):1112-4
references.pmid: 20126730
references.citation: Toxicol Lett. 2011 Feb 5;200(3):201-10
references.pmid: 21130147
references.citation: Appl Microbiol Biotechnol. 2012 Feb;93(3):941-63
references.pmid: 22170106
references.citation: Nano Lett. 2010 Feb 10;10(2):490-3
references.pmid: 20044841
references.citation: J Am Chem Soc. 2008 Aug 20;130(33):10876-7
references.pmid: 18661992
references.citation: J Nanosci Nanotechnol. 2013 Mar;13(3):2091-8
references.pmid: 23755651
references.citation: Free Radic Biol Med. 2006 Apr 1;40(7):1227-33
references.pmid: 16545691
references.citation: ACS Nano. 2012 Apr 24;6(4):2904-16
references.pmid: 22385391
references.citation: J Nanosci Nanotechnol. 2007 Feb;7(2):584-92
references.pmid: 17450800
references.citation: Science. 2008 Jul 18;321(5887):385-8
references.pmid: 18635798
references.citation: Spectrochim Acta A Mol Biomol Spectrosc. 2009 Jul 15;73(2):374-81
references.pmid: 19324587
references.citation: Nano Lett. 2010 Sep 8;10(9):3318-23
references.pmid: 20684528
references.citation: Int J Nanomedicine. 2013;8:4399-413
references.pmid: 24265551
references.citation: ACS Nano. 2010 Jul 27;4(7):4317-23
references.pmid: 20593851
references.citation: Int J Nanomedicine. 2013;8:413-20
references.pmid: 23378763
references.citation: Int J Nanomedicine. 2012;7:5901-14
references.pmid: 23226696
references.citation: Nanomedicine (Lond). 2013 Oct;8(10):1669-88
references.pmid: 24074389
references.citation: Nat Nanotechnol. 2008 Feb;3(2):101-5
references.pmid: 18654470