Galactosylated low molecular weight chitosan as DNA carrier for hepatocyte-targeting.
Chitosan has the potential for DNA complexation and is useful as a non-viral vector for gene delivery. Highly purified low molecular weight chitosan (LMWC) was prepared. Lactobionic acid (LA) bearing galactose group was coupled with LMWC for liver-specificity. A series of galactosylated-LMWC (gal-LMWC) samples covering a range of galactose group contents were prepared. The chitosan/DNA complexes were obtained using a complex coacervation process. Gal-LMWCs were used to transfer pSV-beta-galactosidase reporter gene into human hepatocellular carcinoma cell (HepG2), L-02, SMMC-7721, and human cervix adenocarcinoma cell line (HeLa) cell lines in vitro. Transfection efficiency of gal-LMWCs was evaluated by beta-galactosidase assay and compared with those of lipofectin, calcium phosphate (CaP), high molecular weigh chitosan (HMWC) and LMWC. Gal-LMWC/DNA complex shows a very efficient cell selective transfection to hepatocyte. The transfection efficiency of gal-LMWCs increased with the improvement of the galactosylation degree. Cytotoxicity of gal-LMWC was determined by 3-(4,5-dimethylthiazd-2-yl)-2,5-diphenyltentrazolium bromide (MTT) assay and the results show that the modified chitosan has relatively low cytotoxicity, giving the evidence that the modified chitosan vector has the potential to be used as a safe gene-delivery system.
Gao
Shuying
S
State Key Laboratory of Pharmaceutical Biotechnology, Department of Biochemistry, Nanjing University, Nanjing 210093, PR China.
Chen
Jiangning
J
Xu
Xuerong
X
Ding
Zhi
Z
Yang
Yong-Hua
YH
Hua
Zichun
Z
Zhang
Junfeng
J
D002130:Calcium Phosphates; D002240:Carbohydrate Sequence; D002686:Chitin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000633:toxicity; D048271:Chitosan; D003460:Crystallization; D004187:Disaccharides / Q000737:chemistry; D004337:Drug Carriers / Q000737:chemistry / Q000633:toxicity; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000008:administration & dosage; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010714:Phosphatidylethanolamines; D014162:Transfection; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D002130:Calcium Phosphates; D004187:Disaccharides; D004337:Drug Carriers; D010714:Phosphatidylethanolamines; C485828:alpha-tricalcium phosphate; C485830:tetracalcium phosphate; D002686:Chitin; C005608:lactobionic acid; C485838:calcium phosphate, monobasic, anhydrous; C020888:1,2-dielaidoylphosphatidylethanolamine; D048271:Chitosan; C020243:calcium phosphate; D001616:beta-Galactosidase; C485829:calcium phosphate, dibasic, anhydrous