Csaba N, Sánchez A, Alonso M J
Department Pharmaceutical Technology, School of Pharmacy, University of Santiago de Compostela, 15782, Santiago, Spain.
J Control Release. 2006 Jun 28;113(2):164-72. doi: 10.1016/j.jconrel.2006.03.017. Epub 2006 Apr 5.
We have recently reported the formation of a new type of nanoparticles consisting of blends of poly (lactic-co-glycolic acid) (PLGA) and polyethylene oxide (PEO) derivatives, which exhibit the capacity to associate and release plasmid DNA in a controlled manner. In the present work our goal was to investigate the ability of these nanoparticles to overcome cellular and mucosal barriers (i.e. nasal mucosa) and thus, to work as gene delivery carriers. First, we studied the in vitro cellular uptake (HEK 293 cell line) of FITC-labelled plasmid DNA nanoencapsulated in PLGA: Pluronic F68 and PLGA: Tetronic T904 particles by confocal microscopy. Second, we investigated the uptake of rhodamine-labelled nanoparticles by the nasal mucosa following intranasal administration to mice. Third, we monitored the immune response generated by the nanoparticles containing a beta-galactosidase encoding gene, following nasal administration to mice, using the ELISA technique. The results of the in vitro cell culture studies showed the ability of these new nanoparticles to enter the cells and transport the associated DNA molecule across the cell membrane. Moreover, the results obtained following in vivo administration of the fluorescent nanoparticles evidenced their capability to overcome the nasal mucosal barrier. Finally, the results of the immunisation studies showed that DNA-loaded nanoparticles elicit a fast and strong response, significantly more pronounced than that corresponding to the naked plasmid DNA for up to 6 weeks. Overall, these results suggest that these new nanoparticles have a potential as carriers for the delivery of DNA across the nasal mucosa.
我们最近报道了一种新型纳米颗粒的形成，该纳米颗粒由聚（乳酸-乙醇酸共聚物）（PLGA）和聚环氧乙烷（PEO）衍生物的混合物组成，它们具有以可控方式结合和释放质粒DNA的能力。在本研究中，我们的目标是研究这些纳米颗粒克服细胞和粘膜屏障（即鼻粘膜）的能力，从而作为基因传递载体发挥作用。首先，我们通过共聚焦显微镜研究了包裹在PLGA: Pluronic F68和PLGA: Tetronic T904颗粒中的FITC标记质粒DNA的体外细胞摄取（HEK 293细胞系）。其次，我们研究了鼻内给药小鼠后鼻粘膜对罗丹明标记纳米颗粒的摄取。第三，我们使用ELISA技术监测了鼻内给药小鼠后含β-半乳糖苷酶编码基因的纳米颗粒产生的免疫反应。体外细胞培养研究结果表明，这些新型纳米颗粒能够进入细胞并将相关的DNA分子转运穿过细胞膜。此外，荧光纳米颗粒体内给药后获得的结果证明了它们克服鼻粘膜屏障的能力。最后，免疫研究结果表明，负载DNA的纳米颗粒引发快速而强烈的反应，在长达6周的时间内，这种反应明显比裸体质粒DNA更显著。总体而言，这些结果表明，这些新型纳米颗粒有潜力作为跨鼻粘膜递送DNA的载体。
