Poly(D,L-lactide-co-glycolide acid) nanoparticles for DNA delivery:: Waiving preparation complexity and increasing efficiency
Poly(D,L-lactide-co-glycolide acid) nanoparticles for DNA delivery:: Waiving preparation complexity and increasing efficiency
复制标题
DOI：
10.1002/bip.20697
复制
发表时间：
2007-04-05
期刊：
影响因子：
2.9
通讯作者：
Machluf, Marcelle
中科院分区：
文献类型：
--
作者：
Gvili, Koby;Benny, Ofra;Machluf, Marcelle
在设计基于聚合物纳米颗粒(NPs)的非病毒基因传递系统时，重要的是要记住与未来临床应用相关的障碍。简化NPs生产程序并考虑毒性是需要解决的最重要的问题。当使用阳离子聚合物/脂类、缓冲剂和蛋白质等添加剂时，临床试验中的毒性问题可能会引起关注。因此，本研究的目的是通过缩短超声等步骤并在保持其效率的同时避免使用添加剂来简化聚丙交酯-乙交酯共聚物纳米粒的配方。采用改进的w/o/w工艺制备了300 nm的NPS，DNA包封率为80%。一旦获得这样的纳米粒，处方参数，如DNA载量、释放动力学、DNA完整性和生物活性、细胞摄取和毒性都得到了解决。这些NPs很容易被几个细胞系摄取，并且主要定位于它们的内溶性室。这些纳米颗粒不影响细胞的活力。最重要的是，在COS-7和Cf2th细胞中的转基因研究结果显示，与对照组相比，蛋白质表达水平增加了250倍。这些表达水平高于使用更复杂的NPs系统获得的表达水平，证明了我们简化的NPs用于基因传递的效率。(C)2007年威利期刊公司。
When designing a nonviral gene delivery system based on polymeric nanoparticles (NPs), it is important to keep in mind obstacles associated with future clinical applications. Simplifying the procedure of NPs production and taking toxicity into account are the most important issues that need to be addressed. Toxicity concerns in clinical trials may be raised when using additives such as cationic polymers/lipids, buffering reagents, and proteins. Therefore, the aim of this study was to simplify the formulation of poly (lactide-co-glycolide) acid NPs by shortening steps such as sonication time and by avoiding the use of additives while preserving its efficiency. NPs (300 nm) were formulated using a modified w/o/w technique with DNA entrapment efficiency of 80%. Once achieving such NPs, formulation parameters such as DNA loading, release kinetics, DNA integrity and bioactivity, uptake by cells, and toxicity were addressed. The NPs were readily taken by several cell lines and were localized mostly in their endolysosornal compartments. The NPs did not affect cells viability. Most importantly, transfection studies in COS-7 and Cf2th cells resulted with a 250-fold protein expression levels when compared with the control. These expression levels are higher than ones achieved with more complication NPs systems, demonstrating the efficiency of our simplified NPs for gene delivery. (c) 2007 Wiley periodicals, Inc.