Lactosylated chitosan for DNA delivery into hepatocytes: the effect of lactosylation on the physicochemical properties and intracellular trafficking of pDNA/chitosan complexes.
Chitosan is a useful nonviral vector for gene delivery. To make a pDNA/chitosan complex specific to hepatocytes, lactose-modified chitosan (lac-chitosan) was synthesized. When the percentage of lactose residues substituted was 8%, lac-chitosan showed excellent DNA-binding ability, good protection of DNA from nuclease, and the suppression of self-aggregation and serum-induced aggregation. Although the cellular uptake efficiency of the pDNA/lac-chitosan complex was almost the same as that of the pDNA/chitosan complex, the cell transfection efficiency of the former was greater for HepG2 cells having asialoglycoprotein receptors. Inhibitor of endocytosis such as bafilomycin A1 and nocodazole significantly reduced the transfection efficiency of the pDNA/lac-chitosan complex. Observations with a confocal laser scanning microscope indicated that the pDNA/lac-chitosan complexes traversed endocytic compartments more rapidly than the pDNA/chitosan complex. Furthermore, the pDNA/lac-chitosan complexes were delivered to the late endosome and have the advantage of delivering DNA to the perinuclear region.
Faculty of Science and Technology, Keio University, 3-14-1 Hiyoshi, Yokohama, 223-8522, Japan.
Mayu
M
Hashimoto
Minoru
M
Morimoto
Hiroyuki
H
Saimoto
Yoshihiro
Y
Shigemasa
Toshinori
T
Sato
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry / Q000378:metabolism; D001692:Biological Transport; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002460:Cell Line; D048271:Chitosan / Q000737:chemistry / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D007785:Lactose / Q000737:chemistry* / Q000378:metabolism; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D015394:Molecular Structure; D014162:Transfection
D001672:Biocompatible Materials; D046911:Macromolecular Substances; D004247:DNA; D048271:Chitosan; D003850:Deoxyribonuclease I; D007785:Lactose