Enhancement of chitosan nanoparticle-facilitated gene transfection by ultrasound both in vitro and in vivo.
In recent years, inefficiency of transfection and the lack of safe gene vectors have limited the feasibility of gene therapy. Fabrication of a vector that is safe and has high transfection efficiency is crucial for the development of successful gene therapies. Herein, we complexed chitosan to plasmids at various N/P ratios, the molar ratios of the amino groups of chitosan to the phosphate groups of DNA, to create chitosan-DNA nanoparticles (CDNs), and then measured CDNs size, zeta-potential, efficiency of plasmid complexation, and plasmid integrity from enzyme digestion. We also used flow cytometry and fluorescence microscopy to examine the effect of an ultrasound (US) regimen on the efficiency of transfection of HeLa cells. The results revealed that the average size, zeta-potential, and loading efficiency of plasmid DNA in CDNs were 180-200 nm, 26-35 mV, and greater than 80%, respectively. Moreover, the transgene expression could be enhanced efficiently while HeLa cells or tumor tissues were given CDNs and then treated with US. Therefore, the use of chitosan nanoparticles and an US regimen shows great promise as an effective method of gene therapy.
Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, Taipei 100, Taiwan.
Shu-Jyuan
SJ
Yang
Szu-Min
SM
Chang
Kun-Che
KC
Tsai
Han-Min
HM
Tsai
Wen-Shiang
WS
Chen
Ming-Jium
MJ
Shieh
D048271:Chitosan / Q000737:chemistry* / Q000494:pharmacology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006801:Humans; D053758:Nanoparticles / Q000737:chemistry*; D010957:Plasmids / Q000737:chemistry* / Q000494:pharmacology; D013016:Sound; D019076:Transgenes
D048271:Chitosan