Effect of chitosan-alginate nanoparticles and ultrasound on the efficiency of gene transfection of human cancer cells.
BACKGROUND
Gene therapy has been used to treat a variety of health problems, but transfection inefficiency and the lack of safe vectors have limited clinical progress. Fabrication of a vector that is safe and has high transfection efficiency is crucial for the development of successful gene therapy. The present study aimed to synthesize chitosan-alginate nanoparticles that can be used as carriers of the pAcGFP1-C1 plasmid and to use these nanoparticles with an ultrasound protocol to achieve high efficiency gene transfection.


METHODS
Chitosan was complexed with alginate and the pAcGFP1-C1 plasmid at different charge ratios to create chitosan-alginate-DNA nanoparticles (CADNs). The average particle size and loading efficiency were measured. Plasmid DNA retardation and integrity were analysed on 1% agarose gels. The effect of CADNs and ultrasound on the efficiency of transfection of cells and subcutaneous tumors was evaluated.


RESULTS
In the CADNs, the average size of incorporated plasmid DNA was 600-650 nm and the loading efficiency was greater than 90%. On the basis of the results of the plasmid DNA protection test, CADNs could protect the transgene from DNase I degradation. The transgene product expression could be enhanced efficiently if cells or tumor tissues were first given CADNs and then treated with ultrasound.


CONCLUSIONS
The use of CADNs combined with an ultrasound regimen is a promising method for safe and effective gene therapy.
Yang
Shu-Jyuan
SJ
Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, Taipei, Taiwan.
Chang
Szu-Min
SM
Tsai
Kun-Che
KC
Chen
Wen-Shiang
WS
Lin
Feng-Huei
FH
Shieh
Ming-Jium
MJ
D000464:Alginates / Q000737:chemistry* / Q000633:toxicity; D000818:Animals; D016923:Cell Death / Q000187:drug effects; D048271:Chitosan / Q000737:chemistry* / Q000633:toxicity; D004247:DNA / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D020723:Glucuronic Acid / Q000737:chemistry / Q000633:toxicity; D006367:HeLa Cells; D006603:Hexuronic Acids / Q000737:chemistry / Q000633:toxicity; D006801:Humans; D051379:Mice; D008856:Microscopy, Fluorescence; D053758:Nanoparticles / Q000737:chemistry* / Q000633:toxicity; D009369:Neoplasms / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D014162:Transfection / Q000379:methods*; D014465:Ultrasonics
D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D004247:DNA; D048271:Chitosan