Dendrimer modified magnetic iron oxide nanoparticle/DNA/PEI ternary magnetoplexes: a novel strategy for magnetofection

As a powerful technology to enhance the efficiency of gene delivery, magnetofection has attracted considerable attention in the past decade. In this work, we introduced 6 generation of PAMAM dendrimer modified superparamagnetic nanoparticles (DMSPION-G6) to PEI/DNA polyplexes by a two-step process and enhanced the transfection efficiency of PEI with the help of a magnetic field. We prepared DMSPION-G6/DNA/PEI ternary magnetoplexes by precondensing DMSPION-G6 with DNA at a low mass ratio to yield DMSPION-G6/DNA magnetoplexes with negative surface charge, followed by further coating with branched PEI (25 kDa) via electrostatic interactions. We measured the transfection efficiencies of DMSPION-G6/DNA/PEI ternary magnetoplexes in COS-7, 293T and HeLa cells in the presence or absence of a magnetic field. Compared with PEI/DNA polyplexes, DMSPION-G6/DNA/PEI ternary magnetoplexes exhibited enhanced transfection efficiencies in all the three cell lines when a magnetic field was applied, especially in the presence of 10% FBS. Time-resolved and dose-resolved transfection indicated that high-level transgene expression was achievable with a relatively short incubation time and low DNA dose when magnetofection was employed. Further evidence from Prussian blue staining, quantification of cellular iron concentration and cellular uptake of Cy-3 labelled DNA demonstrated that the magnetic field could quickly gather the magnetoplexes to the surface of target cells and consequently enhanced the uptake of magnetoplexes by the cells. This represents a novel strategy for polycation-based in vitrogene delivery enhanced by a magnetic field.

[1]  C. Ozkan,et al.  Dendrimer-modified magnetic nanoparticles enhance efficiency of gene delivery system. , 2007, Cancer research.

[2]  Norio Tada,et al.  A novel magnetic crystal-lipid nanostructure for magnetically guided in vivo gene delivery. , 2009, Nature nanotechnology.

[3]  Leaf Huang,et al.  Non-viral is superior to viral gene delivery. , 2007, Journal of controlled release : official journal of the Controlled Release Society.

[4]  Won Jong Kim,et al.  Synergistic effect of low cytotoxic linear polyethylenimine and multiarm polyethylene glycol: study of physicochemical properties and in vitro gene transfection. , 2009, Molecular pharmaceutics.

[5]  Beatriz Pelaz,et al.  The effect of static magnetic fields and tat peptides on cellular and nuclear uptake of magnetic nanoparticles. , 2010, Biomaterials.

[6]  Mark A. Kay,et al.  Progress and problems with the use of viral vectors for gene therapy , 2003, Nature Reviews Genetics.

[7]  H. Gu,et al.  Control of aggregate size of polyethyleneimine-coated magnetic nanoparticles for magnetofection , 2009 .

[8]  D. Scherman,et al.  A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[9]  Alain Pluen,et al.  Delivery of therapeutic shRNA and siRNA by Tat fusion peptide targeting BCR-ABL fusion gene in Chronic Myeloid Leukemia cells. , 2010, Journal of controlled release : official journal of the Controlled Release Society.

[10]  T. Young,et al.  Multilayered polyplexes with the endosomal buffering polycation in the core and the cell uptake-favorable polycation in the outer layer for enhanced gene delivery. , 2010, Biomaterials.

[11]  J Henke,et al.  Magnetofection: enhancing and targeting gene delivery by magnetic force in vitro and in vivo , 2002, Gene Therapy.

[12]  Muhammet S. Toprak,et al.  BSA immobilization on amine-functionalized superparamagnetic iron oxide nanoparticles , 2004 .

[13]  Suzie H Pun,et al.  Extracellular barriers to in Vivo PEI and PEGylated PEI polyplex-mediated gene delivery to the liver. , 2008, Bioconjugate chemistry.

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

[15]  W. Mark Saltzman,et al.  Enhancement of transfection by physical concentration of DNA at the cell surface , 2000, Nature Biotechnology.

[16]  Y. Kaneda,et al.  Magnetic nanoparticles with surface modification enhanced gene delivery of HVJ-E vector. , 2005, Biochemical and biophysical research communications.

[17]  C. Pellegrino,et al.  Efficient transfection of DNA or shRNA vectors into neurons using magnetofection , 2007, Nature Protocols.

[18]  N. M. Moore,et al.  Characterization of a multifunctional PEG-based gene delivery system containing nuclear localization signals and endosomal escape peptides. , 2009, Acta biomaterialia.

[19]  Meredith A Mintzer,et al.  Nonviral vectors for gene delivery. , 2009, Chemical reviews.

[20]  M. Weitzman,et al.  Gene therapy: twenty-first century medicine. , 2005, Annual review of biochemistry.

[21]  Hirokazu Akiyama,et al.  Genetically engineered angiogenic cell sheets using magnetic force-based gene delivery and tissue fabrication techniques. , 2010, Biomaterials.

[22]  K. Leong,et al.  Galactosylated ternary DNA/polyphosphoramidate nanoparticles mediate high gene transfection efficiency in hepatocytes. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[23]  R. Zhuo,et al.  Transfection and intracellular trafficking characteristics for poly(amidoamine)s with pendant primary amine in the delivery of plasmid DNA to bone marrow stromal cells. , 2009, Biomaterials.

[24]  Jindrich Kopecek,et al.  Prospects for cationic polymers in gene and oligonucleotide therapy against cancer. , 2002, Advanced drug delivery reviews.

[25]  M. Hashida,et al.  In vitro cytotoxicity of macromolecules in different cell culture systems , 1995 .

[26]  Jong-sang Park,et al.  PAMAM-PEG-PAMAM: novel triblock copolymer as a biocompatible and efficient gene delivery carrier. , 2004, Biomacromolecules.

[27]  In-Kyu Park,et al.  Hybrid superparamagnetic iron oxide nanoparticle-branched polyethylenimine magnetoplexes for gene transfection of vascular endothelial cells. , 2010, Biomaterials.

[28]  Luigi Naldini,et al.  Viral vectors for gene therapy: the art of turning infectious agents into vehicles of therapeutics , 2001, Nature Medicine.

[29]  Daniel W. Pack,et al.  Design and development of polymers for gene delivery , 2005, Nature Reviews Drug Discovery.

[30]  Mattias Belting,et al.  Nuclear delivery of macromolecules: barriers and carriers. , 2005, Advanced drug delivery reviews.

[31]  J. Rosenecker,et al.  The Magnetofection Method: Using Magnetic Force to Enhance Gene Delivery , 2003, Biological chemistry.

[32]  A S Hoffman,et al.  A pH-sensitive polymer that enhances cationic lipid-mediated gene transfer. , 2001, Bioconjugate chemistry.

[33]  J. Suh,et al.  Gene delivery to differentiated neurotypic cells with RGD and HIV Tat peptide functionalized polymeric nanoparticles. , 2006, Biomaterials.

[34]  J. Dobson,et al.  Polyethyleneimine functionalized iron oxide nanoparticles as agents for DNA delivery and transfection , 2007 .

[35]  R. Zhuo,et al.  Novel poly(amidoamine)s with pendant primary amines as highly efficient gene delivery vectors. , 2010, Macromolecular bioscience.

[36]  J. Rosenecker,et al.  Insights into the mechanism of magnetofection using PEI‐based magnetofectins for gene transfer , 2004, The journal of gene medicine.

[37]  A. Urtti,et al.  Extracellular and intracellular barriers in non-viral gene delivery. , 2003, Journal of controlled release : official journal of the Controlled Release Society.

[38]  J. Xue,et al.  Gene transfer using self-assembled ternary complexes of cationic magnetic nanoparticles, plasmid DNA and cell-penetrating Tat peptide. , 2010, Biomaterials.

[39]  P. Choong,et al.  Selective gene delivery for cancer therapy using cationic liposomes: in vivo proof of applicability. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[40]  J. Rosenecker,et al.  Gene delivery to respiratory epithelial cells by magnetofection , 2004, The journal of gene medicine.

[41]  Yu Zhang,et al.  Protective coating of superparamagnetic iron oxide nanoparticles , 2003 .

[42]  S. Zahler,et al.  Magnetofection--a highly efficient tool for antisense oligonucleotide delivery in vitro and in vivo. , 2003, Molecular therapy : the journal of the American Society of Gene Therapy.

[43]  W. Fan,et al.  Investigation of polyethylenimine‐grafted‐triamcinolone acetonide as nucleus‐targeting gene delivery systems , 2010, The journal of gene medicine.

[44]  T. Niidome,et al.  Gene Therapy Progress and Prospects: Nonviral vectors , 2002, Gene Therapy.

[45]  D. Luo,et al.  A self-assembled, modular DNA delivery system mediated by silica nanoparticles. , 2004, Journal of controlled release : official journal of the Controlled Release Society.