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.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3961286 | PMC |
| http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0092076 | PLOS |
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