Center of Genomics and Bioinformatics Research, Faculty of Science, Prince of Songkla University, Hat-Yai, Songkhla, 90112, Thailand.
J Mater Sci Mater Med. 2010 May;21(5):1553-61. doi: 10.1007/s10856-010-3993-9. Epub 2010 Jan 26.
An efficient non-viral gene delivery for varieties of cells has been considered essential for gene therapy and tissue engineering. This study evaluated transfection efficiency of chitosan (HW) with molecular weights (Mw) at 470 and degree of deacetylation (DDA) 80% and its depolymerization product (LW) with Mw at 16 kDa and DDA 54%, as well as epidermal growth factor (EGF) conjugated to chitosan-DNA microparticles of both HW and LW by using either disulfide linkage or NHS-PEO(4)-Maleimide as a cross linker. The results revealed that the depolymerized LW at chitosan/DNA charge ratio 56:1 and pH 6.9 gave high transfection efficiency in both KB, a cancer cell line, and fibroblast cells at about the same level of Lipofectamine, but the EGF-conjugated chitosan-DNA polyplexes from these methods did not improve transfection efficiency, which may come from the aggregation and fusing of the complexes as shown in scanning electron microscopy. However, this depolymerized LW chitosan showed the potential for further development as a safe and cost-effective non-viral gene delivery vehicle.
高效的非病毒基因传递已被认为是基因治疗和组织工程的关键。本研究评估了分子量（Mw）为 470 和脱乙酰度（DDA）为 80%的壳聚糖（HW）及其解聚产物（Mw 为 16 kDa 和 DDA 为 54%的 LW）以及表皮生长因子（EGF）与壳聚糖-DNA 微粒的转染效率，使用二硫键或 NHS-PEO(4)-马来酰亚胺作为交联剂。结果表明，在壳聚糖/DNA 电荷比为 56:1 和 pH 值为 6.9 时，解聚的 LW 在 KB（一种癌细胞系）和成纤维细胞中的转染效率均很高，与 Lipofectamine 相当，但这些方法得到的 EGF 缀合壳聚糖-DNA 聚集体并未提高转染效率，这可能是由于扫描电子显微镜显示复合物的聚集和融合。然而，这种解聚的 LW 壳聚糖具有进一步开发为安全、经济高效的非病毒基因传递载体的潜力。
