Biotransformation and Biological Interaction of Graphene and Graphene Oxide during Simulated Oral Ingestion.

The biotransformation and biological impact of few layer graphene (FLG) and graphene oxide (GO) are studied, following ingestion as exposure route. An in vitro digestion assay based on a standardized operating procedure (SOP) is exploited. The assay simulates the human ingestion of nanomaterials during their dynamic passage through the different environments of the gastrointestinal tract (salivary, gastric, intestinal). Physical-chemical changes of FLG and GO during digestion are assessed by Raman spectroscopy. Moreover, the effect of chronic exposure to digested nanomaterials on integrity and functionality of an in vitro model of intestinal barrier is also determined according to a second SOP. These results show a modulation of the aggregation state of FLG and GO nanoflakes after experiencing the complex environments of the different digestive compartments. In particular, chemical doping effects are observed due to FLG and GO interaction with digestive juice components. No structural changes/degradation of the nanomaterials are detected, suggesting that they are biopersistent when administered by oral route. Chronic exposure to digested graphene does not affect intestinal barrier integrity and is not associated with inflammation and cytotoxicity, though possible long-term adverse effects cannot be ruled out.

[1]  I. Yu,et al.  Twenty-Eight-Day Oral Toxicity, Genotoxicity, and Gender-Related Tissue Distribution of Silver Nanoparticles in Sprague-Dawley Rats , 2008 .

[2]  Bing Yan,et al.  Size-dependent cell uptake of protein-coated graphene oxide nanosheets. , 2012, ACS applied materials & interfaces.

[3]  Shoushan Fan,et al.  The dependence of graphene Raman D-band on carrier density. , 2013, Nano letters.

[4]  F. Stellacci,et al.  A general mechanism for intracellular toxicity of metal-containing nanoparticles† †Electronic supplementary information (ESI) available. See DOI: 10.1039/c4nr01234h Click here for additional data file. , 2014, Nanoscale.

[5]  K. Novoselov,et al.  Exploring the Interface of Graphene and Biology , 2014, Science.

[6]  A. Ferrari,et al.  Doping dependence of the Raman spectrum of defected graphene. , 2014, ACS nano.

[7]  L. Hoffmann,et al.  Effects of silver nanoparticles and ions on a co-culture model for the gastrointestinal epithelium , 2015, Particle and Fibre Toxicology.

[8]  E. Vázquez,et al.  Interaction of graphene-related materials with human intestinal cells: an in vitro approach. , 2016, Nanoscale.

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

[10]  H. R. Krishnamurthy,et al.  Monitoring dopants by Raman scattering in an electrochemically top-gated graphene transistor. , 2008, Nature nanotechnology.

[11]  Andre K. Geim,et al.  The rise of graphene. , 2007, Nature materials.

[12]  H. Bouwmeester,et al.  Behaviour of silver nanoparticles and silver ions in an in vitro human gastrointestinal digestion model , 2012, Nanotoxicology.

[13]  A. Kraegeloh,et al.  Penetration of CdSe/ZnS quantum dots into differentiated vs undifferentiated Caco-2 cells , 2016, Journal of Nanobiotechnology.

[14]  Bengt Fadeel,et al.  Classification framework for graphene-based materials. , 2014, Angewandte Chemie.

[15]  A. Ferrari,et al.  Raman spectroscopy of graphene and graphite: Disorder, electron phonon coupling, doping and nonadiabatic effects , 2007 .

[16]  Delyan R. Hristov,et al.  Low uptake of silica nanoparticles in Caco-2 intestinal epithelial barriers , 2017, Beilstein journal of nanotechnology.

[17]  Sundara Ramaprabhu,et al.  A Raman spectroscopic investigation of graphite oxide derived graphene , 2012 .

[18]  K. Novoselov,et al.  Breakdown of the adiabatic Born-Oppenheimer approximation in graphene. , 2007, Nature materials.

[19]  Agnes G. Oomen,et al.  Presence of nano-sized silica during in vitro digestion of foods containing silica as a food additive. , 2012, ACS nano.

[20]  Peter Wick,et al.  Uptake of label-free graphene oxide by Caco-2 cells is dependent on the cell differentiation status , 2017, Journal of Nanobiotechnology.

[21]  A. Geim,et al.  Unconventional quantum Hall effect and Berry’s phase of 2π in bilayer graphene , 2006, cond-mat/0602565.

[22]  A. Galeone,et al.  Negligible particle-specific toxicity mechanism of silver nanoparticles: the role of Ag+ ion release in the cytosol. , 2015, Nanomedicine : nanotechnology, biology, and medicine.

[23]  Azlin Mustapha,et al.  Toxicity of graphene oxide on intestinal bacteria and Caco-2 cells. , 2015, Journal of food protection.

[24]  P. Artursson,et al.  Correlation between oral drug absorption in humans and apparent drug permeability coefficients in human intestinal epithelial (Caco-2) cells. , 1991, Biochemical and biophysical research communications.

[25]  Agnes B Kane,et al.  Biological interactions of graphene-family nanomaterials: an interdisciplinary review. , 2012, Chemical research in toxicology.

[26]  A. Dignass,et al.  Intestinal barrier function , 2002, Current opinion in clinical nutrition and metabolic care.

[27]  Domschke,et al.  IL‐4, IL‐10 and IL‐13 down‐regulate monocyte‐chemoattracting protein‐1 (MCP‐1) production in activated intestinal epithelial cells , 1998, Clinical and experimental immunology.

[28]  J. Heyder,et al.  Deposition of particles in the human respiratory tract in the size range 0.005–15 μm , 1986 .

[29]  S. Santucci,et al.  Flake size-dependent cyto and genotoxic evaluation of graphene oxide on in vitro A549, CaCo2 and vero cell lines. , 2014, Journal of biological regulators and homeostatic agents.

[30]  Judith Klein-Seetharaman,et al.  Carbon nanotubes degraded by neutrophil myeloperoxidase induce less pulmonary inflammation. , 2010, Nature nanotechnology.

[31]  Paolo A Netti,et al.  Transport across the cell-membrane dictates nanoparticle fate and toxicity: a new paradigm in nanotoxicology. , 2014, Nanoscale.

[32]  Cinzia Casiraghi,et al.  Raman study on defective graphene: Effect of the excitation energy, type, and amount of defects , 2013 .

[33]  Andrew D Maynard,et al.  Exposure Assessment Approaches for Engineered Nanomaterials , 2010, Risk analysis : an official publication of the Society for Risk Analysis.

[34]  Weidong Ruan,et al.  Observation of Enhanced Raman Scattering for Molecules Adsorbed on TiO2 Nanoparticles: Charge-Transfer Contribution , 2008 .

[35]  Kai Yang,et al.  In vitro and in vivo behaviors of dextran functionalized graphene , 2011 .

[36]  Andre K. Geim,et al.  Raman spectrum of graphene and graphene layers. , 2006, Physical review letters.

[37]  A. Oomen,et al.  Applicability of an in vitro digestion model in assessing the bioaccessibility of mycotoxins from food. , 2005, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[38]  C. Casiraghi.  Doping dependence of the Raman peaks intensity of graphene close to the Dirac point , 2009, 0908.4480.

[39]  John Robertson,et al.  Interpretation of infrared and Raman spectra of amorphous carbon nitrides , 2003 .

[40]  A. Ferrari,et al.  Graphene Photonics and Optoelectroncs , 2010, CLEO 2012.

[41]  N. Pugno,et al.  Fragmentation and exfoliation of 2-dimensional materials: a statistical approach. , 2014, Nanoscale.

[42]  Haifang Wang,et al.  Biological effect of food additive titanium dioxide nanoparticles on intestine: an in vitro study , 2015, Journal of applied toxicology : JAT.

[43]  Lang Tran,et al.  A unified framework for nanosafety is needed , 2014 .

[44]  Sang-Jae Kim,et al.  The chemical and structural analysis of graphene oxide with different degrees of oxidation , 2013 .

[45]  Wei Yang,et al.  Inhaled nanoparticles--a current review. , 2008, International journal of pharmaceutics.

[46]  S. Stankovich,et al.  Chemical analysis of graphene oxide films after heat and chemical treatments by X-ray photoelectron and Micro-Raman spectroscopy , 2009 .

[47]  A. Grill.  Porous pSiCOH Ultralow-k Dielectrics for Chip Interconnects Prepared by PECVD , 2009 .

[48]  L. Brinson,et al.  Functionalized graphene sheets for polymer nanocomposites. , 2008, Nature nanotechnology.

[49]  Wei Wei,et al.  The role of the lateral dimension of graphene oxide in the regulation of cellular responses. , 2012, Biomaterials.

[50]  Fabio Benfenati,et al.  Graphene Oxide Nanosheets Disrupt Lipid Composition, Ca(2+) Homeostasis, and Synaptic Transmission in Primary Cortical Neurons. , 2016, ACS nano.

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

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

[53]  Robert H. Hurt,et al.  All in the graphene family - A recommended nomenclature for two-dimensional carbon materials , 2013 .

[54]  J. Laureyns,et al.  Comparative performance of X-ray diffraction and Raman microprobe techniques for the study of carbon materials , 1998 .

[55]  L. Novotný,et al.  Raman characterization of defects and dopants in graphene , 2015, Journal of physics. Condensed matter : an Institute of Physics journal.

[56]  J. Klein-Seetharaman,et al.  The enzymatic oxidation of graphene oxide. , 2011, ACS nano.

[57]  Raimo Hartmann,et al.  Temperature: the "ignored" factor at the NanoBio interface. , 2013, ACS nano.

[58]  M. Ema,et al.  A review of toxicity studies on graphene‐based nanomaterials in laboratory animals , 2017, Regulatory toxicology and pharmacology : RTP.

[59]  Pasquale Bove,et al.  Dissolution test for risk assessment of nanoparticles: a pilot study. , 2017, Nanoscale.

[60]  W. D. de Jong,et al.  Nano-silver – a review of available data and knowledge gaps in human and environmental risk assessment , 2009 .

[61]  K. Novoselov,et al.  Born-Oppenheimer Breakdown in Graphene , 2006, cond-mat/0611714.

[62]  R. Ruoff,et al.  Graphene, related two-dimensional crystals, and hybrid systems for energy conversion and storage , 2015, Science.

[63]  Vasileios Koutsos,et al.  Minimal oxidation and inflammogenicity of pristine graphene with residence in the lung , 2014, Nanotoxicology.

[64]  Lin Zhao,et al.  Protein corona mitigates the cytotoxicity of graphene oxide by reducing its physical interaction with cell membrane. , 2015, Nanoscale.

[65]  C. Casiraghi,et al.  Tunable D peak in gated graphene , 2014, Nano Research.

[66]  Kirsten Gerloff,et al.  Influence of simulated gastrointestinal conditions on particle-induced cytotoxicity and interleukin-8 regulation in differentiated and undifferentiated Caco-2 cells , 2013, Nanotoxicology.

[67]  S. Nguyen,et al.  Crumpled Graphene Nanosheets as Highly Effective Barrier Property Enhancers , 2010, Advanced materials.

[68]  Vincenzo Palermo,et al.  Dispersibility-Dependent Biodegradation of Graphene Oxide by Myeloperoxidase. , 2015, Small.

[69]  J. Tour,et al.  Effective drug delivery, in vitro and in vivo, by carbon-based nanovectors noncovalently loaded with unmodified Paclitaxel. , 2010, ACS nano.

[70]  A. Bianco.  Graphene: Safe or Toxic? The Two Faces of the Medal , 2013 .

[71]  R. Car,et al.  Raman spectra of graphite oxide and functionalized graphene sheets. , 2008, Nano letters.

[72]  M. Prato,et al.  Exfoliation of graphite with triazine derivatives under ball-milling conditions: preparation of few-layer graphene via selective noncovalent interactions. , 2014, ACS nano.

[73]  Marco P Monopoli,et al.  Biomolecular coronas provide the biological identity of nanosized materials. , 2012, Nature nanotechnology.

[74]  Jun Ma,et al.  Studies on the properties of graphene oxide-reinforced starch biocomposites , 2011 .

[75]  K. Novoselov,et al.  Raman Fingerprint of Charged Impurities in Graphene , 2007, 0709.2566.

[76]  C. Hierold,et al.  Raman imaging of doping domains in graphene on SiO2 , 2007, 0709.4156.

[77]  Claudia Röhl,et al.  Manufactured nanomaterials: categorization and approaches to hazard assessment , 2014, Archives of Toxicology.

[78]  M. Kruszewski,et al.  Silver nanoparticles induced changes in the expression of NF-κB related genes are cell type specific and related to the basal activity of NF-κB. , 2014, Toxicology in vitro : an international journal published in association with BIBRA.

[79]  M. Dresselhaus,et al.  Defect characterization in graphene and carbon nanotubes using Raman spectroscopy , 2010, Philosophical Transactions of the Royal Society A: Mathematical, Physical and Engineering Sciences.

[80]  Iseult Lynch,et al.  How safe are nanomaterials? , 2015, Science.

[81]  P. Netti,et al.  Shuttle-mediated nanoparticle delivery to the blood-brain barrier. , 2013, Small.

[82]  Alfonso Lampen,et al.  Analytically monitored digestion of silver nanoparticles and their toxicity on human intestinal cells , 2014, Nanotoxicology.

[83]  Giuseppe Iannaccone,et al.  Electronics based on two-dimensional materials. , 2014, Nature nanotechnology.