Zhao Xia, Yu Shi-Bing, Wu Feng-Lan, Mao Ze-Bin, Yu Chang-Long
The Department of Pharmaceutics, Peking University School of Pharmaceutical Sciences, Beijing 100083, China.
J Control Release. 2006 May 15;112(2):223-8. doi: 10.1016/j.jconrel.2006.01.016. Epub 2006 Mar 23.
Chitosan-pEGFP nanoparticles were synthesized through the complex coacervation of the cationic polymer with pEGFP, in order to examine the potential of chitosan as a non-viral gene delivery vector to transfer exogenous gene into primary chondrocytes for the treatment of joint diseases. The nanoparticles were prepared at an N/P ratio of 3.8 and showed a spherical or irregular shape. The mean particle size and zeta potential of the nanoparticles freshly prepared with chitosan of different molecular weight were in the range of 100-300 nm and varied from +1 to +23 mV, respectively. Both the particle size and the zeta potential altered in DMEM of different pH. The transfection of primary chondrocytes was performed in different conditions by varying pH of transfection medium, molecular weight of chitosan and different plasmid dosage. Analysis of FACS demonstrated that the transfection efficiency could reach a much high level and the percentage of positive cells could exceed 50% in certain condition. These results suggest that chitosan-DNA nanoparticles have favorable characteristics for non-viral gene delivery to primary chondrocytes, and have the potential to deliver therapeutic genes directly into joint.
壳聚糖-pEGFP纳米粒子通过阳离子聚合物与pEGFP的复凝聚法合成，以研究壳聚糖作为非病毒基因递送载体将外源基因导入原代软骨细胞用于治疗关节疾病的潜力。纳米粒子在N/P比为3.8的条件下制备，呈球形或不规则形状。用不同分子量壳聚糖新制备的纳米粒子的平均粒径和zeta电位分别在100-300nm范围内和+1至+23mV之间变化。在不同pH值的DMEM中，粒径和zeta电位均发生改变。通过改变转染培养基的pH值、壳聚糖的分子量和不同的质粒剂量，在不同条件下对原代软骨细胞进行转染。流式细胞术分析表明，转染效率可达到很高水平，在某些条件下阳性细胞百分比可超过50%。这些结果表明，壳聚糖-DNA纳米粒子具有将非病毒基因递送至原代软骨细胞的良好特性，并且具有将治疗性基因直接递送至关节的潜力。
