Dose-dependent effects of nanoscale graphene oxide on reproduction capability of mammals

Abstract In vivo dose-dependent effects of nanoscale graphene oxide (NGO) sheets on reproduction capability of Balb/C mice were investigated. Biodistribution study of the NGO sheets (intravenously injected into male mice at dose of ∼2000 μg/mL or 4 mg/kg of body weight) showed a high graphene uptake in testis. Hence, in vivo effects of the NGO sheets on important characteristics of spermatozoa (including their viability, morphology, kinetics, DNA damage and chromosomal aberration) were evaluated. Significant in vivo effects was found at the injected concentrations ≥200 μg/mL after (e.g., ∼45% reduction in sperm viability and motility at 2000 μg/mL). Observation of remarkable DNA fragmentations and chromosomal aberrations of the spermatozoa after ∼8 weeks from the first weekly injection were assigned to the involvement of the NGO in spermatogenesis of the mice. The uptake of the NGO in the testis could also increase the generation of reactive oxygen species in semen of the mice. Moreover, semen of the NGO-treated mice (containing the damaged spermatozoa) might disturb the hormone secretion and pregnant functionality of female mice (∼44, 35 and 59% reduction in fertility, gestation ability and multi-production capability) and also viability of the next generation (∼15% reduction in postnatal viability of delivered pups).

[1]  M. Mahmoudi,et al.  Graphene: promises, facts, opportunities, and challenges in nanomedicine. , 2013, Chemical reviews.

[2]  O. Akhavan,et al.  Spongy graphene electrode in electrochemical detection of leukemia at single-cell levels , 2014 .

[3]  Omid Akhavan,et al.  Zinc ferrite spinel-graphene in magneto-photothermal therapy of cancer. , 2014, Journal of materials chemistry. B.

[4]  T. Seo,et al.  A Controllable Self‐Assembly Method for Large‐Scale Synthesis of Graphene Sponges and Free‐Standing Graphene Films , 2010 .

[5]  O. Akhavan,et al.  Near infrared laser stimulation of human neural stem cells into neurons on graphene nanomesh semiconductors. , 2015, Colloids and surfaces. B, Biointerfaces.

[6]  K. Tremellen.  Oxidative stress and male infertility--a clinical perspective. , 2008, Human reproduction update.

[7]  Akira Shinohara,et al.  Rad51‐deficient vertebrate cells accumulate chromosomal breaks prior to cell death , 1998, The EMBO journal.

[8]  Xin Wang,et al.  Graphene−Metal Particle Nanocomposites , 2008 .

[9]  Kai Yang,et al.  In vivo biodistribution and toxicology of functionalized nano-graphene oxide in mice after oral and intraperitoneal administration. , 2013, Biomaterials.

[10]  Kai Yang,et al.  The influence of surface chemistry and size of nanoscale graphene oxide on photothermal therapy of cancer using ultra-low laser power. , 2012, Biomaterials.

[11]  C. Hierold,et al.  Spatially resolved Raman spectroscopy of single- and few-layer graphene. , 2006, Nano letters.

[12]  Roberto Car,et al.  Functionalized single graphene sheets derived from splitting graphite oxide. , 2006, The journal of physical chemistry. B.

[13]  Zhijun Zhang,et al.  Functional graphene oxide as a nanocarrier for controlled loading and targeted delivery of mixed anticancer drugs. , 2010, Small.

[14]  O. Akhavan,et al.  DNA and RNA extractions from eukaryotic and prokaryotic cells by graphene nanoplatelets , 2014 .

[15]  Liangzhu Feng,et al.  Graphene in biomedicine: opportunities and challenges. , 2011, Nanomedicine.

[16]  O. Akhavan,et al.  Protein Degradation and RNA Efflux of Viruses Photocatalyzed by Graphene–Tungsten Oxide Composite Under Visible Light Irradiation , 2012 .

[17]  H. Emamy,et al.  Genotoxicity of graphene nanoribbons in human mesenchymal stem cells , 2013 .

[18]  B. Müller-Hill,et al.  Mutant bZip‐DNA complexes with four quasi‐identical protein‐DNA interfaces. , 1996, The EMBO journal.

[19]  O. Akhavan,et al.  Superparamagnetic zinc ferrite spinel–graphene nanostructures for fast wastewater purification , 2014 .

[20]  Fengbo Li,et al.  Toxicology of Graphene Oxide Nanosheets Against Paecilomyces catenlannulatus , 2015, Bulletin of Environmental Contamination and Toxicology.

[21]  Christy L Haynes,et al.  Cytotoxicity of graphene oxide and graphene in human erythrocytes and skin fibroblasts. , 2011, ACS applied materials & interfaces.

[22]  Qing Huang,et al.  Influence of polyethylene glycol coating on biodistribution and toxicity of nanoscale graphene oxide in mice after intravenous injection , 2014, International journal of nanomedicine.

[23]  O. Akhavan,et al.  Toward single-DNA electrochemical biosensing by graphene nanowalls. , 2012, ACS nano.

[24]  R. Car,et al.  Single Sheet Functionalized Graphene by Oxidation and Thermal Expansion of Graphite , 2007 .

[25]  Omid Akhavan,et al.  Graphene nanomesh promises extremely efficient in vivo photothermal therapy. , 2013, Small.

[26]  Zhuang Liu,et al.  PEGylated nanographene oxide for delivery of water-insoluble cancer drugs. , 2008, Journal of the American Chemical Society.

[27]  O. Akhavan,et al.  Wrapping bacteria by graphene nanosheets for isolation from environment, reactivation by sonication, and inactivation by near-infrared irradiation. , 2011, The journal of physical chemistry. B.

[28]  Keyvan Bijanzad,et al.  Synthesis of graphene from natural and industrial carbonaceous wastes , 2014 .

[29]  Chwee Teck Lim,et al.  Origin of enhanced stem cell growth and differentiation on graphene and graphene oxide. , 2011, ACS nano.

[30]  J. Robertson,et al.  Interpretation of Raman spectra of disordered and amorphous carbon , 2000 .

[31]  O. Akhavan.  Graphene nanomesh by ZnO nanorod photocatalysts. , 2010, ACS nano.

[32]  O. Akhavan,et al.  Accelerated differentiation of neural stem cells into neurons on ginseng-reduced graphene oxide sheets , 2014 .

[33]  Moon Gyu Sung,et al.  Enhanced Differentiation of Human Neural Stem Cells into Neurons on Graphene , 2011, Advanced materials.

[34]  N. Durán,et al.  Redox‐enzymes, cells and micro‐organisms acting on carbon nanostructures transformation: A mini‐review , 2013, Biotechnology progress.

[35]  M. Suh,et al.  The control of neural cell-to-cell interactions through non-contact electrical field stimulation using graphene electrodes. , 2011, Biomaterials.

[36]  Omid Akhavan,et al.  Photocatalytic Reduction of Graphene Oxide Nanosheets on TiO2 Thin Film for Photoinactivation of Bacteria in Solar Light Irradiation , 2009 .

[37]  Omid Akhavan,et al.  Size-dependent genotoxicity of graphene nanoplatelets in human stem cells. , 2012, Biomaterials.

[38]  Kwang S. Kim,et al.  Large-scale pattern growth of graphene films for stretchable transparent electrodes , 2009, Nature.

[39]  O. Akhavan,et al.  The use of graphene in the self-organized differentiation of human neural stem cells into neurons under pulsed laser stimulation. , 2014, Journal of materials chemistry. B.

[40]  Yanli Chang,et al.  In vitro toxicity evaluation of graphene oxide on A549 cells. , 2011, Toxicology letters.

[41]  Kai Yang,et al.  Multimodal Imaging Guided Photothermal Therapy using Functionalized Graphene Nanosheets Anchored with Magnetic Nanoparticles , 2012, Advanced materials.

[42]  O. Akhavan,et al.  Increasing the antioxidant activity of green tea polyphenols in the presence of iron for the reduction of graphene oxide , 2012 .

[43]  C. Gagnon,et al.  Involvement of reactive oxygen species in human sperm arcosome reaction induced by A23187, lysophosphatidylcholine, and biological fluid ultrafiltrates. , 1998, Journal of andrology.

[44]  O. Akhavan.  Bacteriorhodopsin as a superior substitute for hydrazine in chemical reduction of single-layer graphene oxide sheets , 2015 .

[45]  C. N. Lau,et al.  Temperature dependence of the Raman spectra of graphene and graphene multilayers. , 2007, Nano letters.

[46]  Linlin Li,et al.  Effects of graphene oxide on the development of offspring mice in lactation period. , 2015, Biomaterials.

[47]  James M. Tour,et al.  Growth of graphene from food, insects, and waste. , 2011, ACS nano.

[48]  O. Akhavan.  The effect of heat treatment on formation of graphene thin films from graphene oxide nanosheets , 2010 .

[49]  Kai Yang,et al.  Graphene in mice: ultrahigh in vivo tumor uptake and efficient photothermal therapy. , 2010, Nano letters.

[50]  Nelson Durán,et al.  Nanotoxicity of graphene and graphene oxide. , 2014, Chemical research in toxicology.

[51]  O. Akhavan,et al.  Differentiation of human neural stem cells into neural networks on graphene nanogrids. , 2013, Journal of materials chemistry. B.

[52]  O. Akhavan,et al.  Ultra-sensitive detection of leukemia by graphene. , 2014, Nanoscale.

[53]  Omid Akhavan,et al.  Toxicity of graphene and graphene oxide nanowalls against bacteria. , 2010, ACS nano.

[54]  Mehdi Shamsara,et al.  Cyto and genotoxicities of graphene oxide and reduced graphene oxide sheets on spermatozoa , 2014 .

[55]  N. Mohanty,et al.  Graphene-based single-bacterium resolution biodevice and DNA transistor: interfacing graphene derivatives with nanoscale and microscale biocomponents. , 2008, Nano letters.

[56]  Yang Xu,et al.  Cytotoxicity effects of graphene and single-wall carbon nanotubes in neural phaeochromocytoma-derived PC12 cells. , 2010, ACS nano.

[57]  C. Fan,et al.  Uniform ultrasmall graphene oxide nanosheets with low cytotoxicity and high cellular uptake. , 2013, ACS applied materials & interfaces.

[58]  Bei Wang,et al.  Synthesis of enhanced hydrophilic and hydrophobic graphene oxide nanosheets by a solvothermal method , 2009 .

[59]  Omid Akhavan,et al.  Flash photo stimulation of human neural stem cells on graphene/TiO2 heterojunction for differentiation into neurons. , 2013, Nanoscale.

[60]  A. Geim,et al.  Two-dimensional gas of massless Dirac fermions in graphene , 2005, Nature.

[61]  Chen Liqiang,et al.  Toxicity of graphene oxide and multi-walled carbon nanotubes against human cells and zebrafish , 2012 .

[62]  R. Tice,et al.  Single cell gel/comet assay: Guidelines for in vitro and in vivo genetic toxicology testing , 2000, Environmental and molecular mutagenesis.

[63]  Elena Stolyarova,et al.  Graphene oxidation: thickness-dependent etching and strong chemical doping. , 2008, Nano letters.

[64]  O. Akhavan,et al.  Graphene nanogrids for selective and fast osteogenic differentiation of human mesenchymal stem cells , 2013 .

[65]  Omid Akhavan,et al.  Escherichia coli bacteria reduce graphene oxide to bactericidal graphene in a self-limiting manner , 2012 .

[66]  Chunhai Fan,et al.  Distribution and biocompatibility studies of graphene oxide in mice after intravenous administration , 2011 .

[67]  Omid Akhavan,et al.  Adverse effects of graphene incorporated in TiO2 photocatalyst on minuscule animals under solar light irradiation , 2012 .

[68]  M. Dresselhaus,et al.  Raman spectroscopy in graphene , 2009 .

[69]  M. I. Katsnelson,et al.  Graphene: New bridge between condensed matter physics and quantum electrodynamics , 2007, cond-mat/0703374.

[70]  K. Novoselov,et al.  Detection of individual gas molecules adsorbed on graphene. , 2006, Nature materials.

[71]  W. Ford,et al.  Glycolysis and sperm motility: does a spoonful of sugar help the flagellum go round? , 2006, Human reproduction update.

[72]  H. Emamy,et al.  Nontoxic concentrations of PEGylated graphene nanoribbons for selective cancer cell imaging and photothermal therapy , 2012 .

[73]  Kai Yang,et al.  In vivo pharmacokinetics, long-term biodistribution, and toxicology of PEGylated graphene in mice. , 2011, ACS nano.

[74]  C. Fan,et al.  Protein corona-mediated mitigation of cytotoxicity of graphene oxide. , 2011, ACS nano.

[75]  A. Irajizad,et al.  Melatonin as a powerful bio-antioxidant for reduction of graphene oxide , 2011 .

[76]  Zhuang Liu,et al.  Nano-graphene oxide for cellular imaging and drug delivery , 2008, Nano research.

[77]  E. Bertoft,et al.  Determination of hormones by time-resolved fluoroimmunoassay. , 1984, Talanta.