Multifunctional Chitosan Magnetic-Graphene (CMG) Nanoparticles: a Theranostic Platform for Tumor-targeted Co-delivery of Drugs, Genes and MRI Contrast Agents.

Combing chemotherapy with gene therapy has been one of the most promising strategies for the treatment of cancer. The noninvasive MRI with superparamagnetic iron oxide (SPIO) as contrast agent is one of the most effecitve techniques for evaluating the antitumor therapy. However, to construct a single system that can deliver efficiently gene, drug and SPIO to the cancer site remains a challenge. Herein, we report a chitosan functionalized magnetic graphene nanoparticle (CMG) platform for simultaneous gene/drug and SPIO delivery to tumor. The phantom and ex vivo MRI images suggest CMG as a strong T2 contrast-enhancing agent. The CMGs are biocompatible as evaluated by the WST assay and predominantly accumulate in tumors as shown by biodistribution studies and MRI. The anticancer drug doxorubicin (DOX) loaded CMGs (DOX-CMGs) release DOX faster at pH 5.1 than at pH 7.4, and more effective (IC50 = 2 μM) in killing A549 lung cancer cells than free DOX (IC50 = 4 μM). CMGs efficiently deliver DNA into A549 lung cancer cells and C42b prostate cancer cells. In addition, i.v. administration of GFP-plasmid encapsulated within DOX-CMGs into tumor-bearing mice has showed both GFP expression and DOX accumulation at the tumor site at 24 and 48 hrs after administration. These results indicate CMGs provide a robust and safe theranostic platform, which integrates targeted delivery of both gene medicine and chemotherapeutic drug(s), and enhanced MR imaging of tumors. The integrated chemo- and gene- therapeutic and diagnostic design of CMG nanoparticles shows promise for simultaneous targeted imaging, drug delivery and real -time monitoring of therapeutic effect for cancer.

[1]  G. Rosen,et al.  Tumor necrosis factor-related apoptosis-inducing ligand and chemotherapy cooperate to induce apoptosis in mesothelioma cell lines. , 2001, American journal of respiratory cell and molecular biology.

[2]  Chang-Sik Ha,et al.  Synthesis and Drug‐Delivery Behavior of Chitosan‐Functionalized Graphene Oxide Hybrid Nanosheets , 2011 .

[3]  M. Ychou,et al.  Assessment of liver metastases from colorectal adenocarcinoma following chemotherapy: SPIO-MRI versus FDG-PET/CT , 2010, La radiologia medica.

[4]  J. Stoker,et al.  Comparison of MRI (including SS SE-EPI and SPIO-enhanced MRI) and FDG-PET/CT for the detection of colorectal liver metastases , 2009, European Radiology.

[5]  Chunhai Fan,et al.  A Graphene Nanoprobe for Rapid, Sensitive, and Multicolor Fluorescent DNA Analysis , 2010 .

[6]  Xiaogang Qu,et al.  Using Graphene Oxide High Near‐Infrared Absorbance for Photothermal Treatment of Alzheimer's Disease , 2012, Advanced materials.

[7]  Ronghua Yang,et al.  Carbon nanotubes protect DNA strands during cellular delivery. , 2008, ACS nano.

[8]  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.

[9]  Jie Huang,et al.  Polyethylenimine-functionalized graphene oxide as an efficient gene delivery vector , 2011 .

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

[11]  Kai Yang,et al.  In vivo targeting and imaging of tumor vasculature with radiolabeled, antibody-conjugated nanographene. , 2012, ACS nano.

[12]  T. Metens,et al.  Focal liver lesion detection and characterization: comparison of non-contrast enhanced and SPIO-enhanced diffusion-weighted single-shot spin echo echo planar and turbo spin echo T2-weighted imaging. , 2009, European journal of radiology.

[13]  M. Xiong,et al.  Combination of TRAIL gene therapy and chemotherapy enhances antitumor and antimetastasis effects in chemosensitive and chemoresistant breast cancers. , 2003, Molecular therapy : the journal of the American Society of Gene Therapy.

[14]  Kian Ping Loh,et al.  Graphene-based SELDI probe with ultrahigh extraction and sensitivity for DNA oligomer. , 2010, Journal of the American Chemical Society.

[15]  Huang-Hao Yang,et al.  Using graphene to protect DNA from cleavage during cellular delivery. , 2010, Chemical communications.

[16]  I-Wei Chen,et al.  Quantum‐Dot‐Tagged Reduced Graphene Oxide Nanocomposites for Bright Fluorescence Bioimaging and Photothermal Therapy Monitored In Situ , 2012, Advanced materials.

[17]  Il-Kwon Oh,et al.  Graphene oxide-polyethylenimine nanoconstruct as a gene delivery vector and bioimaging tool. , 2011, Bioconjugate chemistry.

[18]  S. Bose,et al.  Chemical functionalization of graphene and its applications , 2012 .

[19]  Yuan Ping,et al.  Chitosan-functionalized graphene oxide as a nanocarrier for drug and gene delivery. , 2011, Small.

[20]  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.

[21]  Ho Sup Yoon,et al.  Co-delivery of drugs and DNA from cationic core–shell nanoparticles self-assembled from a biodegradable copolymer , 2006, Nature materials.

[22]  Tae Seok Seo,et al.  Graphene oxide arrays for detecting specific DNA hybridization by fluorescence resonance energy transfer. , 2010, Biosensors & bioelectronics.

[23]  Debabrata Dash,et al.  Amine-modified graphene: thrombo-protective safer alternative to graphene oxide for biomedical applications. , 2012, ACS nano.

[24]  R. Dijkhuizen,et al.  Imaging Neuroinflammation after Stroke: Current Status of Cellular and Molecular MRI Strategies , 2012, Cerebrovascular Diseases.

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

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

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

[28]  Shuhong Yu,et al.  Water-soluble magnetic-functionalized reduced graphene oxide sheets: in situ synthesis and magnetic resonance imaging applications. , 2010, Small.

[29]  S. Stankovich,et al.  Graphene-based composite materials , 2006, Nature.

[30]  Zheng-Rong Lu,et al.  Targeted intracellular codelivery of chemotherapeutics and nucleic acid with a well-defined dendrimer-based nanoglobular carrier. , 2009, Biomaterials.

[31]  Shumeng Liu,et al.  Engineered polyethylenimine/graphene oxide nanocomposite for nuclear localized gene delivery , 2012 .

[32]  H. Olin,et al.  Carbon nanomaterials as drug carriers: Real time drug release investigation , 2012 .

[33]  T. Park,et al.  Co-delivery of siRNA and paclitaxel into cancer cells by biodegradable cationic micelles based on PDMAEMA-PCL-PDMAEMA triblock copolymers. , 2010, Biomaterials.

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

[35]  Xiaojun Cai,et al.  Multifunctional nanocomposite based on graphene oxide for in vitro hepatocarcinoma diagnosis and treatment. , 2012, Journal of biomedical materials research. Part A.

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

[37]  Christian Plank,et al.  Generation of magnetic nonviral gene transfer agents and magnetofection in vitro , 2007, Nature Protocols.

[38]  M. Paul,et al.  6-Mercaptopurine and Daunorubicin Double Drug Liposomes—Preparation, Drug-Drug Interaction and Characterization , 2005, Journal of liposome research.

[39]  Ganesh Gollavelli,et al.  Multi-functional graphene as an in vitro and in vivo imaging probe. , 2012, Biomaterials.

[40]  M. Ishitobi,et al.  SPIO-Enhanced Magnetic Resonance Imaging for the Detection of Metastases in Sentinel Nodes Localized by Computed Tomography Lymphography in Patients with Breast Cancer , 2011, Annals of surgical oncology.

[41]  Huang-Hao Yang,et al.  A graphene platform for sensing biomolecules. , 2009, Angewandte Chemie.

[42]  Xin Huang,et al.  Multi-functionalized graphene oxide based anticancer drug-carrier with dual-targeting function and pH-sensitivity , 2011 .

[43]  S. Mohapatra,et al.  Dual-purpose magnetic micelles for MRI and gene delivery. , 2012, Journal of controlled release : official journal of the Controlled Release Society.

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

[45]  Zhuang Liu,et al.  Graphene based gene transfection. , 2011, Nanoscale.

[46]  T. Minko,et al.  Co-delivery of siRNA and an anticancer drug for treatment of multidrug-resistant cancer. , 2008, Nanomedicine.

[47]  Xiang Gao,et al.  Nonviral gene delivery: What we know and what is next , 2007, The AAPS Journal.

[48]  Anjan Nan,et al.  Combination Drug Delivery Approaches in Metastatic Breast Cancer , 2012, Journal of drug delivery.

[49]  Jiali Zhang,et al.  Biocompatibility of Graphene Oxide , 2010, Nanoscale research letters.

[50]  Mansoor M. Amiji,et al.  Evaluations of combination MDR-1 gene silencing and paclitaxel administration in biodegradable polymeric nanoparticle formulations to overcome multidrug resistance in cancer cells , 2009, Cancer Chemotherapy and Pharmacology.