Karimi Mahdi, Avci Pinar, Mobasseri Rezvan, Hamblin Michael R, Naderi-Manesh Hossein
Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
Wellman Center for Photomedicine, Massachusetts General Hospital, Boston, MA 02114, USA.
J Nanopart Res. 2013 Apr 1;15(4):1651. doi: 10.1007/s11051-013-1651-0.
Natural polymers and proteins such as chitosan (CS) and albumin (Alb) have recently attracted much attention both in drug delivery and gene delivery. The underlying rationale is their unique properties such as biodegradability, biocompatibility and controlled release. This study aimed to prepare novel albumin-chitosan-DNA (Alb-CS-DNA) core-shell nanoparticles as a plasmid delivery system and find the best conditions for their preparation. Phase separation method and ionic interaction were used for preparation of Alb nanoparticles and Alb-CS-DNA core-shell nanoparticles, respectively. The effects of three important independent variables (1) CS/Alb mass ratio, (2) the ratios of moles of the amine groups of cationic polymers to those of the phosphate groups of DNA (N/P ratio), and (3) Alb concentration, on the nanoparticle size and loading efficiency of the plasmid were investigated and optimized through Box-Behnken design of response surface methodology (RSM). The optimum conditions were found to be CS/Alb mass ratio = 3, N/P ratio = 8.24 and Alb concentration = 0.1 mg/mL. The most critical factors for the size of nanoparticles and loading efficiency were Alb concentration and N/P ratio. The optimized nanoparticles had an average size of 176 ± 3.4 nm and loading efficiency of 80 ± 3.9 %. Cytotoxicity experiments demonstrated that the prepared nanoparticles were not toxic. The high cellular uptake of nanoparticles (~85 %) was shown by flow cytometry and fluorescent microscopy.
天然聚合物和蛋白质，如壳聚糖（CS）和白蛋白（Alb），近年来在药物递送和基因递送方面备受关注。其潜在原理是它们具有诸如生物可降解性、生物相容性和控释等独特性质。本研究旨在制备新型白蛋白 - 壳聚糖 - DNA（Alb - CS - DNA）核壳纳米粒子作为质粒递送系统，并找到其制备的最佳条件。分别采用相分离法和离子相互作用制备Alb纳米粒子和Alb - CS - DNA核壳纳米粒子。通过响应面法（RSM）的Box - Behnken设计，研究并优化了三个重要自变量（1）CS/Alb质量比、（2）阳离子聚合物胺基与DNA磷酸基团的摩尔比（N/P比）以及（3）Alb浓度对纳米粒子大小和质粒负载效率的影响。发现最佳条件为CS/Alb质量比 = 3、N/P比 = 8.24和Alb浓度 = 0.1 mg/mL。影响纳米粒子大小和负载效率的最关键因素是Alb浓度和N/P比。优化后的纳米粒子平均大小为176 ± 3.4 nm，负载效率为80 ± 3.9 %。细胞毒性实验表明所制备的纳米粒子无毒。流式细胞术和荧光显微镜显示纳米粒子具有较高的细胞摄取率（约85 %）。
