Department of Hospital Pharmacy, Nagasaki University Hospital, 1-7-1 Sakamoto, Nagasaki, Japan.
J Pharm Pharm Sci. 2010;13(3):351-61. doi: 10.18433/j3gk52.
In this study, we developed various ternary complexes of encapsulated polyplexes and lipoplexes using chondroitin sulfate (CS) and investigated their universal usefulness for gene delivery.
To prepare the cationic complexes, pDNA was mixed with some cationic vectors such as poly-L-arginine, poly-L-lysine, N-[1-(2, 3-dioleyloxy) propyl]-N, N, N-trimethylammonium chloride (DOTMA)-cholesterol liposomes, and DOTMA- dioleylphosphatidylethanolamine (DOPE) liposomes. CS was added to the cationic complexes for constructions of ternary complexes. We examined in vitro transfection efficiency, cytotoxicity, hematotoxicity, and in vivo transfection efficiency of the ternary complexes.
The cationic polymers and cationic liposomes bound to pDNA and formed stable cationic polyplexes and lipoplexes, respectively. Those cationic complexes showed high transgene efficiency in B16-F10 cells; however, they also had high cytotoxicity and strong agglutination with erythrocytes. CS could encapsulate the polyplexes and lipoplexes and form stable anionic particles without disrupting their structures. The ternary complexes encapsulated by CS showed high transgene efficiency in B16-F10 cells with low cytotoxicity and agglutination. As the result of animal experiments, the polyplexes had little transgene efficiency after intravenous administration in mice, whereas polyplexes encapsulated by CS showed specifically high transgene efficiency in the spleen. The capsulation of CS, however, reduced the high transgene efficiency of the lipoplexes.
These results indicate that CS can contribute to polyplex-mediated gene delivery systems for effective and safe gene therapy.
在这项研究中，我们使用硫酸软骨素（CS）开发了各种包封的聚合物复合物和脂质体的三元复合物，并研究了它们在基因传递中的普遍适用性。
为了制备阳离子复合物，将 pDNA 与一些阳离子载体如聚精氨酸、聚赖氨酸、N-[1-(2,3-二油烯氧基)丙基]-N, N, N-三甲基氯化铵（DOTMA）-胆固醇脂质体和 DOTMA-二油酰基磷脂酰乙醇胺（DOPE）脂质体混合。CS 被添加到阳离子复合物中以构建三元复合物。我们研究了三元复合物的体外转染效率、细胞毒性、血液毒性和体内转染效率。
阳离子聚合物和阳离子脂质体与 pDNA 结合，分别形成稳定的阳离子聚合物复合物和脂质体复合物。这些阳离子复合物在 B16-F10 细胞中显示出高转染效率；然而，它们也具有高细胞毒性和与红细胞强烈聚集。CS 可以包裹聚合物复合物和脂质体，并形成稳定的阴离子颗粒，而不会破坏其结构。CS 包裹的三元复合物在 B16-F10 细胞中显示出高转染效率，同时具有低细胞毒性和低聚集性。动物实验的结果表明，聚合物复合物在静脉注射后在小鼠体内的转染效率较低，而 CS 包裹的聚合物复合物在脾脏中表现出特异性的高转染效率。然而，CS 的包裹降低了脂质体的高转染效率。
这些结果表明，CS 可以为有效的和安全的基因治疗提供聚合物介导的基因传递系统。
