Musashino University, Research Institute of Pharmaceutical Sciences, Tokyo, Japan.
J Pharm Biomed Anal. 2010 Jan 5;51(1):268-72. doi: 10.1016/j.jpba.2009.07.024. Epub 2009 Jul 25.
We have developed a new type of ternary gene transfection system comprising plasmid, polyethyleneimine, and hyaluronic acid (HA). HA recharged the positively charged DNA complexes into negative, and diminished the adverse interactions with bio-components. HA also improved the transcription efficiency, stabilized the dispersion, and protected the particles against lyophilizing induced inactivation. It enabled the preparation of very small complex particles, which showed high in vivo gene expression. In spite of the many attractive properties of the ternary complex systems, their detailed structure is still not clear. In this study, surface structures of the complexes were analyzed by non-confocal laser Raman microscopy. The correlation between the zeta-potential and the Raman spectrum is discussed. The DNA complex that showed the highest gene-expression level was prepared at the mixing ratio of COOH/NH=0.5. At this ratio, all the amino groups were just protonated, and the surface was completely covered by HA attributed to the reduced nonspecific interaction of the complex in which all the amino groups have already interacted with carboxyls of HA, but not too much HA on the surface taken up by the cells.
我们开发了一种新型的三元基因转染系统，包括质粒、聚乙烯亚胺和透明质酸（HA）。HA 使带正电荷的 DNA 复合物带负电，并减少了与生物成分的不良相互作用。HA 还提高了转录效率，稳定了分散性，并保护颗粒免受冻干诱导的失活。它能够制备非常小的复合颗粒，这些颗粒表现出很高的体内基因表达。尽管三元复合物系统具有许多吸引人的特性，但它们的详细结构仍不清楚。在这项研究中，通过非共焦激光拉曼显微镜分析了复合物的表面结构。讨论了 ζ 电位和拉曼光谱之间的相关性。在 COOH/NH=0.5 的混合比下制备了表现出最高基因表达水平的 DNA 复合物。在这个比例下，所有的氨基都被质子化，表面完全被 HA 覆盖，这归因于复合物的非特异性相互作用减少，所有的氨基都已经与 HA 的羧基相互作用，但细胞表面没有太多的 HA 被吸收。
