title: Hydrophilic/lipophilic N-methylene phosphonic chitosan as a promising non-viral vector for gene delivery.
abstract: Cationic amphiphilic drugs have recently been shown to inhibit receptor recycling by disrupting the assembly-disassembly of clathrin at the plasma membrane and endosomes. It is therefore proposed that amphiphilic and cationic polysaccharide macromolecule, when used as gene delivery vectors, may have potential ability to direct the disassembly process of cell membrane organization, and penetrate across the cell membrane into cell and nucleus. In the current study, N-methylene phosphonic chitosan (NMPCS), an amphiphilic macromolecule, was synthesized by incorporating the methylene phosphonic group into the amino groups of chitosan (CS) using formaldehyde as the coupling agent, and characterized with a FTIR spectrometer. NMPCS/DNA or CS/DNA complexes were prepared using a complex coacervation method, and characterized by agarose gel electrophoresis retardation assay and dynamic light scattering (DLS). MTT assay was employed to evaluate the cytotoxicity of the polymers and pGL3-control luciferase plasmid was utilized as a reporter gene to assess the transgenic efficacy of the polymers. It was demonstrated that NMPCS was able to fully entrap the DNA at N/P ratio of 2:1, whereas CS entrapped the DNA completely at N/P ratio of 1:1. DLS showed that the NMPCS/DNA or CS/DNA complexes were of mean diameters ranging from 110 to 180 nm. Neither NMPCS nor CS induced significant loss of cell viability at the concentrations ranging from 1 to 50 microg/ml, whereas PEI at 5 microg/ml started to result in significantly decreased cell viability. The expression of transgene mediated by NMPCS was much higher (more than 100-folds) than that mediated by CS, indicating that NMPCS was a more efficacious gene ferrying vector than CS.
authors.affiliation: Tianjin Key Laboratory of Biomedical Materials, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, China.
authors.forename: Dunwan
authors.initials: D
authors.lastname: Zhu
authors.forename: Kangde
authors.initials: K
authors.lastname: Yao
authors.forename: Jingen
authors.initials: J
authors.lastname: Bo
authors.forename: Hailing
authors.initials: H
authors.lastname: Zhang
authors.forename: Lanxia
authors.initials: L
authors.lastname: Liu
authors.forename: Xia
authors.initials: X
authors.lastname: Dong
authors.forename: Liping
authors.initials: L
authors.lastname: Song
authors.forename: Xigang
authors.initials: X
authors.lastname: Leng
mesh_terms: D019342:Acetic Acid / Q000737:chemistry; D048271:Chitosan / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D004247:DNA / Q000008:administration & dosage / Q000737:chemistry; D004526:Efficiency; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008954:Models, Biological; D053758:Nanoparticles / Q000737:chemistry; D010757:Phosphorous Acids / Q000737:chemistry; D013499:Surface Properties; D013501:Surface-Active Agents / Q000138:chemical synthesis / Q000737:chemistry; D014162:Transfection / Q000379:methods*; D019076:Transgenes; D014780:Viruses
chemical_list: C549127:N-methylene phosphonic chitosan; D010757:Phosphorous Acids; D013501:Surface-Active Agents; D004247:DNA; D048271:Chitosan; D019342:Acetic Acid
references.citation: Nat Med. 1999 Apr;5(4):387-91
references.pmid: 10202926
references.citation: Science. 2002 Mar 29;295(5564):2418-21
references.pmid: 11923529
references.citation: J Control Release. 2008 Sep 10;130(2):129-38
references.pmid: 18538436
references.citation: J Control Release. 2003 Dec 12;93(3):389-402
references.pmid: 14644588
references.citation: Int J Pharm. 2008 Nov 19;364(1):127-34
references.pmid: 18775484
references.citation: Science. 2002 Mar 29;295(5564):2400-3
references.pmid: 11923524
references.citation: J Control Release. 2001 Feb 23;70(3):399-421
references.pmid: 11182210
references.citation: Int J Pharm. 2003 Apr 14;255(1-2):57-68
references.pmid: 12672602
references.citation: J Mater Sci Mater Med. 2009 Mar;20(3):673-80
references.pmid: 18949535
references.citation: Bioconjug Chem. 2002 Jul-Aug;13(4):845-54
references.pmid: 12121141
references.citation: Int J Pharm. 2006 Jun 6;315(1-2):158-66
references.pmid: 16632285
references.citation: Nat Biotechnol. 2003 Aug;21(8):885-90
references.pmid: 12833071
references.citation: Biomaterials. 2005 May;26(14):2147-56
references.pmid: 15576190
references.citation: Proc Natl Acad Sci U S A. 2003 Aug 5;100(16):9138-43
references.pmid: 12886020
references.citation: Proc Natl Acad Sci U S A. 2002 Nov 12;99(23):14640-5
references.pmid: 12403826
references.citation: Gene Ther. 2004 Jun;11(12):1001-10
references.pmid: 14985789
references.citation: J Biol Chem. 1995 May 19;270(20):12117-22
references.pmid: 7744860
references.citation: Drug Dev Ind Pharm. 1998 Nov;24(11):979-93
references.pmid: 9876553
references.citation: Science. 2002 Aug 9;297(5583):967-73
references.pmid: 12169723
references.citation: Biomaterials. 2006 Mar;27(9):2060-5
references.pmid: 16202449
references.citation: Biomacromolecules. 2001 Winter;2(4):1161-8
references.pmid: 11777388
references.citation: Science. 2001 Jul 6;293(5527):58-9
references.pmid: 11441170
references.citation: Biomaterials. 2001 Aug;22(15):2075-80
references.pmid: 11432586
references.citation: Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2006 Aug;28(4):486-91
references.pmid: 16995298
references.citation: Pharm Res. 1998 Sep;15(9):1326-31
references.pmid: 9755881