State Key Laboratory of Genetic Engineering, Department of Pharmaceutical Sciences, School of Life Sciences, Fudan University, Shanghai 200433, China.
J Control Release. 2010 May 21;144(1):46-54. doi: 10.1016/j.jconrel.2010.01.022. Epub 2010 Jan 20.
Trimethyl chitosan-cysteine conjugate (TMC-Cys) was evaluated as non-viral gene carriers to combine the advantages of TMC and thiolated chitosan. TMC-Cys with various molecular weights (30, 100, and 200 kDa) and quaternization degrees (15 and 30%) was allowed to form polyelectrolyte nanocomplexes with plasmid encoding enhanced green fluorescence protein (pEGFP), which demonstrated preferable diameters of below 200 nm and zeta potentials of +15 to +20 mV. Cell binding and uptake of TMC-Cys/pEGFP nanocomplexes (TMC-Cys NC) were enhanced 2.4-3.0 and 1.4-3.0 folds, respectively, compared to TMC/pEGFP nanocomplexes (TMC NC). pEGFP could be easily released from TMC-Cys NC at the intracellular glutathione concentration, which promoted its nuclear transport and accumulation. Consequently, TMC-Cys NC showed a 1.4 to 3.2-fold increase in the transfection efficiency in HEK293 cells as compared to TMC NC and the optimal TMC-Cys(100,30) NC showed a 1.5-fold enhancement than Lipofectamine2000. Such results were further confirmed by in vivo transfection with a 2.3-fold and 4.1-fold higher transfection efficiency of TMC-Cys(100,30) NC than TMC(100,30) NC and Lipofectamine2000, respectively. Therefore, TMC-Cys/DNA nanocomplexes could be a promising gene delivery system with in vitro and in vivo superiority to Lipofectamine2000.
三甲基壳聚糖-半胱氨酸缀合物（TMC-Cys）被评估为非病毒基因载体，以结合 TMC 和巯基化壳聚糖的优势。具有不同分子量（30、100 和 200 kDa）和季铵化度（15 和 30%）的 TMC-Cys 与编码增强型绿色荧光蛋白（pEGFP）的质粒形成聚电解质纳米复合物，其直径小于 200nm，zeta 电位为+15 至+20mV。与 TMC/pEGFP 纳米复合物（TMC NC）相比，TMC-Cys/pEGFP 纳米复合物（TMC-Cys NC）的细胞结合和摄取分别增强了 2.4-3.0 倍和 1.4-3.0 倍。在细胞内谷胱甘肽浓度下，pEGFP 可以很容易地从 TMC-Cys NC 中释放出来，从而促进其核转运和积累。因此，与 TMC NC 相比，TMC-Cys NC 在 HEK293 细胞中的转染效率提高了 1.4 至 3.2 倍，最佳的 TMC-Cys（100,30）NC 比 Lipofectamine2000 提高了 1.5 倍。体内转染实验进一步证实了这一结果，与 TMC（100,30）NC 和 Lipofectamine2000 相比，TMC-Cys（100,30）NC 的转染效率分别提高了 2.3 倍和 4.1 倍。因此，TMC-Cys/DNA 纳米复合物可能是一种有前途的基因传递系统，具有优于 Lipofectamine2000 的体外和体内优势。
