title: Ultrasound treatment increases transfection efficiency of low molecular weight chitosan in fibroblasts but not in KB cells.
abstract: The aim of this study was to optimize transfection efficiency (TE) of the depolymerized low molecular weight (LW) chitosan with molecular weight (Mw) at 16 kDa and 54% degree of deacetylation (DDA) on three primary cells of fibroblast (F), dental pulp (P), and periodontal ligament (PDL). The effect of low frequency ultrasound treatment on the chitosan-DNA complexes prior transfection on TE was also evaluated. This LW chitosan required high N/P ratio (>34) to bind DNA completely. An N/P ratio above 56 tended to improve TE in most primary cells nearly at the level of Lipofectamine. Ultrasonication can reduce the aggregation and sizes of the chitosan-DNA microparticles. It increased TE of F cells at an N/P ratio above 34, which was higher than Lipofectamine. However, this ultrasound treatment caused loss of TE in KB cells. MTT assay of these chitosan-DNA complexes revealed no significant cytotoxicity to both KB and F cells. This LW chitosan has potential for further development into a safer alternative to gene delivery systems in various cells of interest; however the optimal conditions have to be adjusted, depending on each cell source.
authors.affiliation: Department of Oral Biology and Occlusion, Faculty of Dentistry, Prince of Songkla University, Hat-Yai, Songkhla, Thailand.
authors.forename: Ureporn
authors.initials: U
authors.lastname: Kedjarune-Leggat
authors.affiliation: Department of Oral Biology and Occlusion, Faculty of Dentistry, Prince of Songkla University, Hat-Yai, Songkhla, Thailand; Department of Molecular Biotechnology and Bioinformatics, Faculty of Science, Prince of Songkla University, Hat-Yai, Songkhla, Thailand.
authors.forename: Chanyapat
authors.initials: C
authors.lastname: Supaprutsakul
authors.affiliation: Department of Molecular Biotechnology and Bioinformatics, Faculty of Science, Prince of Songkla University, Hat-Yai, Songkhla, Thailand.
authors.forename: Wilaiwan
authors.initials: W
authors.lastname: Chotigeat
mesh_terms: D000293:Adolescent; D000328:Adult; D016923:Cell Death; D002460:Cell Line; D048271:Chitosan / Q000737:chemistry* / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D003782:Dental Pulp / Q000166:cytology; D004587:Electrophoresis, Agar Gel; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D007624:KB Cells; D008156:Luciferases / Q000378:metabolism; D008297:Male; D008863:Microspheres; D008875:Middle Aged; D008970:Molecular Weight; D010513:Periodontal Ligament / Q000166:cytology; D055672:Static Electricity; D014162:Transfection / Q000379:methods*; D014465:Ultrasonics
chemical_list: D004247:DNA; D048271:Chitosan; D008156:Luciferases