Characteristics of DEAE-dextran-MMA graft copolymer as a nonviral gene carrier.
A stable and soapless latex of diethylaminoethyl-dextran-methyl methacrylate (DEAE-dextran-MMA) graft copolymer (DDMC) has been developed for nonviral gene delivery vectors that are possible to autoclave. DDMC relatively easily formed a polyion complex between DNA and DDMC by the hydrophobic force of graft poly(MMA) depending on its large positive entropy change (DeltaS). DDMC has been confirmed as having a high protection facility for DNase by DNase degradation test.Transfection activity was determined using the beta-galactosidase assay, and a higher value of 16 times or more was confirmed for the DDMC samples in comparison with one of the starting DEAE-dextran hydrochloride samples. The resulting DDMC, having an amphiphilic domain so as to form a polymer micelle, should become a stable latex with a hydrophilic-hydrophobic microseparated domain. The complex of DDMC and plasmid DNA may be formed on the spherical structure of the amphiphilic microseparated domain of DDMC and have a good affinity to the cell membrane. The infrared absorption spectrum shift to a high-energy direction at around 3450 cm(-1), because of the complexes between DNA and DDMC, may cause the formation of more compact structures, not only by a coulomb force between the phosphoric acid of DNA and the DEAE group of DEAE-dextran copolymer but also by a force from the multi-intermolecule hydrogen bond in the backbone polymer DEAE-dextran and a hydrophobic force from the graft poly(MMA) in DDMC. It is thus concluded that DNA condensation may possibly have a high transfection efficiency via DDMC. The high efficiency of this graft copolymer, which is sterilized by an autoclave, may thus make it a valuable tool for safe gene delivery.
Onishi
Yasuhiko
Y
School of Engineering, Nippon Bunri University, Oita, Japan. vyx00545@nifty.ne.jp
Eshita
Yuki
Y
Murashita
Aya
A
Mizuno
Masaaki
M
Yoshida
Jun
J
D000818:Animals; D002460:Cell Line; D003637:DEAE-Dextran / Q000737:chemistry*; D004247:DNA / Q000008:administration & dosage / Q000737:chemistry* / Q000493:pharmacokinetics*; D004058:Diffusion; D004337:Drug Carriers / Q000737:chemistry*; D004339:Drug Compounding / Q000379:methods; D006801:Humans; D007668:Kidney / Q000378:metabolism*; D008422:Materials Testing; D020366:Methylmethacrylate / Q000737:chemistry*; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure; D010316:Particle Size; D011108:Polymers / Q000737:chemistry; D012474:Salmon; D014162:Transfection / Q000379:methods*; D014780:Viruses / Q000235:genetics
D004337:Drug Carriers; D011108:Polymers; D020366:Methylmethacrylate; D004247:DNA; D003637:DEAE-Dextran