Ionov Maksim, Garaiova Zuzana, Waczulikova Iveta, Wróbel Dominika, Pędziwiatr-Werbicka Elżbieta, Gomez-Ramirez Rafael, de la Mata Francisco Javier, Klajnert Barbara, Hianik Tibor, Bryszewska Maria
Department of General Biophysics, University of Lodz, Lodz, Poland.
Biochim Biophys Acta. 2012 Sep;1818(9):2209-16. doi: 10.1016/j.bbamem.2012.04.019. Epub 2012 May 2.
One of the major limitations in gene therapy is an inability of naked siRNA to passively diffuse through negatively charged cell membranes. Therefore, the siRNA transport into a cell requires efficient carriers. In this work we analyzed the charge-dependent interaction of the complexes of cationic carbosilane dendrimers (CBD) and anti-HIV siRNA (dendriplexes) with the model membranes - large unilamellar vesicles (LUV). We used the second generation of branched with CBD carbon-silicon bonds (CBD-CS) which are water-stable and that of oxygen-silicon bonds (CBD-OS) which are slowly hydrolyzed in aqueous solutions. The LUVs were composed of zwitterionic dimyristoylphosphatidylcholine (DMPC), negatively charged dipalmitoylphosphatidylglycerol (DPPG) and their mixture (DMPC/DPPG, molar ratio 7:3). The interaction of dendriplexes with LUVs affected both zeta potential and size of the vesicles. The changes of these values were larger for the negatively charged LUV. CBD-CS resulted in the decrease of zeta potential values to more negative ones, whereas an opposite effect took place for CBD-OS suggesting a different kind of interaction between LUVs and the dendriplexes. The results indicate that both CBD-CS and CBD-OS can be used for transport of siRNA into the cells. However, CBD-CS are preferred due to a better stability in water and improved bioavailability of siRNA on their surface.
基因治疗的主要局限之一是裸露的小干扰RNA（siRNA）无法被动扩散穿过带负电荷的细胞膜。因此，将siRNA转运到细胞中需要高效的载体。在这项工作中，我们分析了阳离子碳硅烷树枝状大分子（CBD）与抗HIV siRNA的复合物（树枝状复合物）与模型膜——大单层囊泡（LUV）之间的电荷依赖性相互作用。我们使用了第二代带有碳硅键（CBD-CS）的支化树枝状大分子，其在水中稳定，以及带有氧硅键（CBD-OS）的树枝状大分子，其在水溶液中会缓慢水解。LUV由两性离子二肉豆蔻酰磷脂酰胆碱（DMPC）、带负电荷的二棕榈酰磷脂酰甘油（DPPG）及其混合物（DMPC/DPPG，摩尔比7:3）组成。树枝状复合物与LUV的相互作用影响了囊泡的zeta电位和大小。对于带负电荷的LUV，这些值的变化更大。CBD-CS导致zeta电位值降低至更负的值，而CBD-OS则产生相反的效果，这表明LUV与树枝状复合物之间存在不同类型的相互作用。结果表明，CBD-CS和CBD-OS都可用于将siRNA转运到细胞中。然而，由于CBD-CS在水中具有更好的稳定性以及其表面siRNA的生物利用度提高，因此更受青睐。
