Enhancement of chitosan nanoparticle-facilitated gene transfection by ultrasound both in vitro and in vivo
Journal
Journal of Biomedical Materials Research - Part B Applied Biomaterials
Journal Volume
100 B
Journal Issue
7
Pages
1746-1754
Date Issued
2012
Author(s)
Abstract
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. ? 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2012. Copyright ? 2012 Wiley Periodicals, Inc.
SDGs
Other Subjects
Amino group; Average size; Chitosan nanoparticles; Enzyme digestion; Gene transfection; Gene vectors; HeLa cell; In-vitro; Loading efficiency; Molar ratio; Phosphate group; Plasmid DNA; Transfection efficiency; Transgene expression; Tumor tissues; DNA; Efficiency; Flow cytometry; Fluorescence microscopy; Gene therapy; Nanoparticles; Tissue; Ultrasonics; Chitosan; chitosan; chitosan DNA nanoparticle; lipofectamine; luciferase; nanoparticle; phosphate; plasmid DNA; unclassified drug; animal experiment; animal model; article; cancer tissue; cell membrane; complex formation; controlled study; cytotoxicity test; embryo; enzyme degradation; female; flow cytometry; fluorescence microscopy; gene expression; genetic transfection; heterozygote; human; human cell; in vitro study; in vivo study; mouse; nonhuman; particle size; plasmid; transgene; ultrasound; zeta potential; Chitosan; Gene Expression; Gene Transfer Techniques; HeLa Cells; Humans; Nanoparticles; Plasmids; Sound; Transgenes
Type
journal article