Yang, S.-J.S.-J.YangChang, S.-M.S.-M.ChangTsai, K.-C.K.-C.TsaiWEN-SHIANG CHENLin, Feng-HueiFeng-HueiLinMING-JIUM SHIEH2020-03-302020-03-3020101099-498Xhttps://www.scopus.com/inward/record.uri?eid=2-s2.0-76049108694&doi=10.1002%2fjgm.1418&partnerID=40&md5=69b9c65564abf8bf6cf659253630f606https://scholars.lib.ntu.edu.tw/handle/123456789/481474Background: 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. Copyright ? 2009 John Wiley & Sons, Ltd.[SDGs]SDG3alginic acid; chitosan; nanocarrier; nanoparticle; plasmid DNA; animal experiment; animal model; article; controlled study; female; fluorescence; gene therapy; genetic transfection; HeLa cell; human; human cell; mouse; nonhuman; particle size; priority journal; subcutaneous tissue tumor; transgene; ultrasound; Alginates; Animals; Cell Death; Chitosan; DNA; Electrophoresis, Agar Gel; Glucuronic Acid; Hela Cells; Hexuronic Acids; Humans; Mice; Microscopy, Fluorescence; Nanoparticles; Neoplasms; Plasmids; Transfection; Ultrasonicsjournal article10.1002/jgm.1418199433422-s2.0-76049108694