Key Laboratory of Smart Drug Delivery, Ministry of Education & PLA, Department of Pharmaceutics, School of Pharmacy, Fudan University, Shanghai 201203, PR China.
Biomaterials. 2012 Jan;33(2):644-58. doi: 10.1016/j.biomaterials.2011.09.076. Epub 2011 Oct 24.
Biomaterials coated polymer/DNA complexes are developed as an efficient non-viral gene delivery system. It is able to circumvent the changes of various biophysical properties of the biomaterials and the corresponding polymer/DNA nanoparticles with covalent linkage. In the present study, we introduced pH-sensitive carboxymethyl poly (l-histidine) (CM-PLH) and poly (β-amino ester) (PbAE) as functional biomaterials to form CM-PLH/PbAE/DNA core-shell ternary complexes system based on electrostatically adsorbed coatings for gene efficient delivery and transfection. The preparation of the complexes was performed self-assembly in 25 mm sodium acetate buffer solution at pH 5.2. The complexes kept stable nano-size, behaving good condensation capacity and low toxicity, even provided a higher transfection efficiency than the binary complexes (PbAE/DNA without CM-PLH) and transfected up to (89.6 ± 4.45) % in HEK293 and (57.1 ± 2.10) % in B16-F10 in vitro. The ternary complexes significantly enhanced their cellular uptake and endosomal escape which were proved by the results that the complexes could evade the endosomal lumen and localize in the nucleus of treated cells visualized under Fluorescence Confocal Microscopy (FCM). The aforementioned results indicated that CM-PLH with pH-sensitive imidazole groups played an important role in enhancing the endosomal escape and transfection efficiency. The in vivo gene transfection confirmed that the ternary complexes with pGL3-promoter as led to effectively deposit at the tumor site by the EPR effect and shown 4 fold higher luciferase expression in B16-F10 tumor than the binary complexes. Consequently, CM-PLH/PbAE/DNA ternary complexes system exhibited significant improvements in transfection efficiency in comparison with non-coated PbAE/DNA both in vitro and in vivo, highlighting their functional prospect. Our approach and the gene delivery system fabrication could potentially be useful for effective gene delivery and therapies to targeted cells.
包被聚合物/DNA 复合物的生物材料被开发为一种有效的非病毒基因传递系统。它能够避免生物材料和相应的聚合物/DNA 纳米颗粒的各种生物物理性质的变化，与共价键连接。在本研究中，我们引入了 pH 敏感的羧甲基聚（L-组氨酸）（CM-PLH）和聚（β-氨基酯）（PbAE）作为功能性生物材料，基于静电吸附包被形成 CM-PLH/PbAE/DNA 核壳三元复合物系统，用于高效基因传递和转染。复合物的制备是在 25mm 醋酸钠缓冲溶液中于 pH 5.2 下自组装完成的。复合物保持稳定的纳米尺寸，表现出良好的凝聚能力和低毒性，甚至比二元复合物（无 CM-PLH 的 PbAE/DNA）提供更高的转染效率，在 HEK293 中达到（89.6±4.45）%，在 B16-F10 中达到（57.1±2.10）%。三元复合物显著增强了细胞摄取和内涵体逃逸，这可以通过荧光共聚焦显微镜（FCM）观察到复合物可以逃避内涵体腔并定位于处理细胞的核中来证明。上述结果表明，具有 pH 敏感咪唑基团的 CM-PLH 在增强内涵体逃逸和转染效率方面发挥了重要作用。体内基因转染证实，具有 pGL3 启动子的三元复合物通过 EPR 效应有效地沉积在肿瘤部位，在 B16-F10 肿瘤中显示出比二元复合物高 4 倍的荧光素酶表达。因此，与非包被的 PbAE/DNA 相比，CM-PLH/PbAE/DNA 三元复合物系统在体外和体内均显著提高了转染效率，突出了其功能前景。我们的方法和基因传递系统的构建有可能对靶向细胞的有效基因传递和治疗有用。
