The effects of coating pDNA/chitosan complexes with chondroitin sulfate on physicochemical characteristics and cell transfection.
pDNA/chitosan complexes have been investigated as promising non-viral vectors for gene delivery. However, an increase in transfection efficiency and enhancement of physicochemical stability are required for their practical use. In this study, chondroitin sulfate (CS) was employed as a coating agent to increase the stability and transfection efficiency of a pDNA/chitosan complex. The pDNA/chitosan/CS ternary complexes formed with six kinds of CSs having different limiting viscosities (0.2-1.6) and sulfation degrees (5.0-7.0%) showed considerable differences in particle size, surface charge, and morphology. Among them, CS having a medium limiting viscosity (0.5-0.6) and a high sulfation degree (6.9%) showed significant enhancements in cell transfection efficiency. Analyses of cellular uptake and intracellular trafficking revealed that increased cellular uptake via macropinocytosis, together with reduced entry into lysosomes, may explain the promotion of transfection efficiency of ternary complexes.
Department of Biosciences and Informatics, Keio University, Yokohama, Kanagawa, Japan.
Kenji
K
Hagiwara
Mitsuhiro
M
Nakata
Yoshiyuki
Y
Koyama
Toshinori
T
Sato
D000818:Animals; D019556:COS Cells; D002417:Cattle; D002460:Cell Line; D045744:Cell Line, Tumor; D048271:Chitosan / Q000737:chemistry*; D002522:Chlorocebus aethiops; D002809:Chondroitin Sulfates / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D018014:Gene Transfer Techniques; D006801:Humans; D018625:Microscopy, Atomic Force / Q000379:methods; D053758:Nanoparticles / Q000737:chemistry; D010873:Pinocytosis; D011134:Polysaccharides / Q000737:chemistry; D012685:Sepharose / Q000737:chemistry; D014162:Transfection; D014783:Viscosity
D011134:Polysaccharides; D002809:Chondroitin Sulfates; D004247:DNA; D012685:Sepharose; D048271:Chitosan