Chitosan-functionalized graphene oxide as a nanocarrier for drug and gene delivery.

The covalent functionalization of graphene oxide (GO) with chitosan (CS) is successfully accomplished via a facile amidation process. The CS-grafted GO (GO-CS) sheets consist of about 64 wt.% CS, which imparts them with a good aqueous solubility and biocompatibility. Additionally, the physicochemical properties of GO-CS are studied. As a novel nanocarrier, GO-CS is applied to load a water-insoluble anticancer drug, camptothecin (CPT), via π-π stacking and hydrophobic interactions. It is demonstrated that GO-CS possesses a superior loading capacity for CPT, and the GO-CS-CPT complexes show remarkably high cytotoxicity in HepG2 and HeLa cell lines compared to the pure drug. At the same time, GO-CS is also able to condense plasmid DNA into stable, nanosized complexes, and the resulting GO-CS/pDNA nanoparticles exhibit reasonable transfection efficiency in HeLa cells at certain nitrogen/phosphate ratios. Therefore, the GO-CS nanocarrier is able to load and deliver both anticancer drugs and genes.

[1]  Lin Li,et al.  Green fabrication of chitosan films reinforced with parallel aligned graphene oxide , 2011 .

[2]  M. Fang,et al.  Single-layer graphene nanosheets with controlled grafting of polymer chains , 2010 .

[3]  Jiachun Feng,et al.  Click chemistry as a route for the immobilization of well-defined polystyrene onto graphene sheets , 2010 .

[4]  In-Yong Kim,et al.  Chitosan and its derivatives for tissue engineering applications. , 2008, Biotechnology advances.

[5]  Yongsheng Chen,et al.  High-Efficiency Loading and Controlled Release of Doxorubicin Hydrochloride on Graphene Oxide , 2008 .

[6]  R. Ruoff,et al.  The chemistry of graphene oxide. , 2010, Chemical Society reviews.

[7]  Yen Wei,et al.  Synthesis of cellulose-graft-poly(N,N-dimethylamino-2-ethyl methacrylate) copolymers via homogeneous ATRP and their aggregates in aqueous media. , 2008, Biomacromolecules.

[8]  Lin Li,et al.  Thermo- and pH-Responsive Association Behavior of Dual Hydrophilic Graft Chitosan Terpolymer Synthesized via ATRP and Click Chemistry , 2010 .

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

[10]  Y Wang,et al.  Chitosan-DNA nanoparticles as gene carriers: synthesis, characterization and transfection efficiency. , 2001, Journal of controlled release : official journal of the Controlled Release Society.

[11]  T. Ogoshi,et al.  Supramolecular polymer networks from hybrid between graphene oxide and per-6-amino-beta-cyclodextrin. , 2010, Chemical communications.

[12]  X. Zhu,et al.  Cholic acid-modified dendritic multimolecular micelles and enhancement of anticancer drug therapeutic efficacy. , 2010, Bioconjugate chemistry.

[13]  Guoliang Zhang,et al.  Deoxygenation of Exfoliated Graphite Oxide under Alkaline Conditions: A Green Route to Graphene Preparation , 2008 .

[14]  S. Stankovich,et al.  Synthesis of graphene-based nanosheets via chemical reduction of exfoliated graphite oxide , 2007 .

[15]  Jiwei Cui,et al.  Encapsulation of Water‐Insoluble Drugs in Polymer Capsules Prepared Using Mesoporous Silica Templates for Intracellular Drug Delivery , 2010, Advanced materials.

[16]  Xinglin Yang,et al.  Exfoliated graphite oxide decorated by PDMAEMA chains and polymer particles. , 2009, Langmuir : the ACS journal of surfaces and colloids.

[17]  Y. Okahata,et al.  Mechanism of cell transfection with plasmid/chitosan complexes. , 2001, Biochimica et biophysica acta.

[18]  T. Bettinger,et al.  Chitosan-Based Vector/DNA Complexes for Gene Delivery: Biophysical Characteristics and Transfection Ability , 1998, Pharmaceutical Research.

[19]  Lin Li,et al.  Interactions between Ionic Surfactants and Polysaccharides in Aqueous Solutions , 2008 .

[20]  P. Veski,et al.  In vivo evaluation of matrix granules containing microcrystalline chitosan as a gel-forming excipient. , 2003, International journal of pharmaceutics.

[21]  J. Abrahams,et al.  A Graphene Oxide˙Streptavidin Complex for Biorecognition – Towards Affinity Purification , 2010 .

[22]  Yizhe Hu,et al.  Synthesis of amphiphilic graphene nanoplatelets. , 2009, Small.

[23]  R. Piner,et al.  Biocompatible, Robust Free‐Standing Paper Composed of a TWEEN/Graphene Composite , 2010, Advanced materials.

[24]  Lin Li,et al.  Thermo-responsive association of chitosan-graft-poly(N-isopropylacrylamide) in aqueous solutions. , 2010, The journal of physical chemistry. B.

[25]  Dongmin Chen,et al.  Synthesis and Solid-State NMR Structural Characterization of 13C-Labeled Graphite Oxide , 2008, Science.

[26]  Y. Okahata,et al.  In vitro gene delivery mediated by chitosan. effect of pH, serum, and molecular mass of chitosan on the transfection efficiency. , 2001, Biomaterials.

[27]  K. Novoselov,et al.  Control of Graphene's Properties by Reversible Hydrogenation: Evidence for Graphane , 2008, Science.

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

[29]  P. J. Ollivier,et al.  Layer-by-Layer Assembly of Ultrathin Composite Films from Micron-Sized Graphite Oxide Sheets and Polycations , 1999 .

[30]  Steven Nutt,et al.  Covalent polymer functionalization of graphene nanosheets and mechanical properties of composites , 2009 .

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

[32]  Liang Zhao,et al.  P-glycoprotein antibody functionalized carbon nanotube overcomes the multidrug resistance of human leukemia cells. , 2010, ACS nano.

[33]  Yuliang Yang,et al.  Single-Walled Carbon Nanotubes Functionalized with High Bonding Density of Polymer Layers and Enhanced Mechanical Properties of Composites , 2007 .

[34]  Jacek Klinowski,et al.  Structure of Graphite Oxide Revisited , 1998 .

[35]  Gilbert C Walker,et al.  Noncovalent engineering of carbon nanotube surfaces by rigid, functional conjugated polymers. , 2002, Journal of the American Chemical Society.

[36]  B. Han,et al.  Synthesis of graphene/polyaniline composite nanosheets mediated by polymerized ionic liquid. , 2010, Chemical communications.

[37]  M. A. Gómez,et al.  Polymeric Modification of Graphene through Esterification of Graphite Oxide and Poly(vinyl alcohol) , 2009 .

[38]  V. B. Morris,et al.  Studies on the condensation of depolymerized chitosans with DNA for preparing chitosan-DNA nanoparticles for gene delivery applications. , 2009, Journal of biomedical materials research. Part B, Applied biomaterials.

[39]  Yuehe Lin,et al.  Aptamer/graphene oxide nanocomplex for in situ molecular probing in living cells. , 2010, Journal of the American Chemical Society.

[40]  Guohua Chen,et al.  pH-Responsive chitosan-mediated graphene dispersions. , 2010, Langmuir : the ACS journal of surfaces and colloids.

[41]  Chun Li,et al.  Non-covalent functionalization of graphene sheets by sulfonated polyaniline. , 2009, Chemical communications.

[42]  Hongkun He,et al.  General Approach to Individually Dispersed, Highly Soluble, and Conductive Graphene Nanosheets Functionalized by Nitrene Chemistry , 2010 .

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

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

[45]  Jun Li,et al.  Functionalization of Chitosan via Atom Transfer Radical Polymerization for Gene Delivery , 2010 .

[46]  Yanwu Zhu,et al.  Polymer Brushes via Controlled, Surface-Initiated Atom Transfer Radical Polymerization (ATRP) from Graphene Oxide. , 2010, Macromolecular rapid communications.

[47]  Lin Li,et al.  Influence of cetyltrimethylammonium bromide on physicochemical properties and microstructures of chitosan-TPP nanoparticles in aqueous solutions. , 2008, Journal of colloid and interface science.

[48]  K. Neoh,et al.  Comb-shaped copolymers composed of hydroxypropyl cellulose backbones and cationic poly((2-dimethyl amino)ethyl methacrylate) side chains for gene delivery. , 2009, Bioconjugate chemistry.

[49]  Gang Liu,et al.  Organo- and Water-Dispersible Graphene Oxide−Polymer Nanosheets for Organic Electronic Memory and Gold Nanocomposites , 2010 .

[50]  E. Kang,et al.  Self-assembled glycol chitosan nanoparticles for the sustained and prolonged delivery of antiangiogenic small peptide drugs in cancer therapy. , 2008, Biomaterials.

[51]  Rodney S. Ruoff,et al.  Effect of Water Vapor on Electrical Properties of Individual Reduced Graphene Oxide Sheets , 2008 .

[52]  Ya‐Ping Sun,et al.  Polymer functionalization and solubilization of carbon nanosheets. , 2009, Chemical communications.

[53]  H. Yoo,et al.  Quantum-dot-assisted fluorescence resonance energy transfer approach for intracellular trafficking of chitosan/DNA complex. , 2008, Acta biomaterialia.

[54]  Min Zhang,et al.  Co-delivery of doxorubicin and Bcl-2 siRNA by mesoporous silica nanoparticles enhances the efficacy of chemotherapy in multidrug-resistant cancer cells. , 2009, Small.

[55]  Xiaoming Yang,et al.  Well-dispersed chitosan/graphene oxide nanocomposites. , 2010, ACS applied materials & interfaces.

[56]  R. Ruoff,et al.  Chemical methods for the production of graphenes. , 2009, Nature nanotechnology.

[57]  Lin Li,et al.  Stepped association of comb‐like and stimuli‐responsive graft chitosan copolymer synthesized using ATRP and active ester conjugation methods , 2009 .

[58]  C. Chen,et al.  Chitosan-poly(ε-caprolactone)-poly(ethylene glycol) graft copolymers: synthesis, self-assembly, and drug release behavior. , 2010, Journal of biomedical materials research. Part A.