pDNA-巯基壳聚糖纳米粒的制备及Box-Behnken效应面法工艺优化  

Preparation of pDNA-Loaded Thiolated Chitosan Nanoparticles and Optimization by Using Box-Behnken Design and Response Surface Method

作　　者：曾皓月 王之 方美娟[3] 王立强 

机构地区：[1]华侨大学生物医学学院,福建泉州362021 [2]兰州军区总医院临床药学科,甘肃兰州730000 [3]厦门大学药学院,福建厦门361000

出　　处：《华侨大学学报（自然科学版）》2017年第5期676-681,共6页Journal of Huaqiao University(Natural Science)

基　　金：国家自然科学基金资助项目(81302652);福建省自然科学基金资助项目(2015J01342)

摘　　要：以巯基壳聚糖(TCS)为基因载体,采用离子交联法制备能用于基因口服研究的质粒DNA-巯基壳聚糖纳米粒(pDNA-TCS-NPs).分别以TCS质量浓度、三聚磷酸钠(TPP)质量浓度、pH值和转速为考察对象,以pDNA-TCS-NPs粒径和Zeta电位为评价指标,采用4因素3水平Box-Behnken效应面法筛选最佳制备工艺,并对其外观形态,包封率等体外性质进行考察.结果表明:TCS质量浓度为0.80mg·mL^(-1),TPP质量浓度为0.65mg·mL^(-1),pH=5.3,转速为2 000r·min^(-1)是最优制备工艺,可制得粒径为(134.21±1.34)nm,Zeta电位为(24.36±0.29)mV,包封率在(80.26±0.56)%,形状规则且分散良好的pDNA-巯基壳聚糖纳米粒;Box-Behnken实验设计可用于预测和优化pDNA-TCS-NP制备工艺优化筛选.Preliminary exploration to prepare plasmid DNA-loaded thiolated chitosan nanoparticles（pDNATCS-NPs）which can be used for gene oral administration was investigated.Using TCS as gene carriers,the pDNA-TCS-NPs were prepared by the ionic cross-linking method.The effects of the concentrations of TCS and TPP,pH value and stirring speed were investigated on the particle diameter and zeta potential of the pDNA-TCS-NPs.In addition,the formula was optimized by Box-Behnken design and response surface method with four factors and three levels,and the physic-chemical properties such as the shape and encapsulation efficiency were also studied.The results showed that the optimal formula was as follows：TCS concentration 0.80mg·mL（-1）,TPP concentration 0.65mg·mL（-1）,pH 5.3,and stirring speed 2 000r·min（-1）.Particle diameter was（134.21±1.34）nm,zeta potential was（24.36±0.29）mV,and efficiency was（80.26±0.56）%.under the optimal conditions.The pDNA-TCS-NPs showed uniform spherical solid particles with regular shape and ideal uniformly dispersion.It′s also confirmed that the Box-Behnken experimental design and response surface method could be used to predict and optimize the pDNA-loaded TCS nanoparticles.

关 键 词：巯基壳聚糖 质粒DNA 纳米粒 BOX-BEHNKEN效应面法 离子交联法 工艺优化 

分 类 号：R944[医药卫生—药剂学]