pDNA-巯基壳聚糖纳米粒的制备及Box-Behnken效应面法工艺优化

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0.80 mg·mL^-1,TPP质量浓度为0.65 mg·mL^-1,pH=5.3,转速为2 000 r·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.

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

Preliminary exploration to prepare plasmid DNA-loaded thiolated chitosan nanoparticles(pDNA-TCS-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 0.80 mg·mL^-1, TPP concentration 0.65 mg·mL^-1, pH 5.3, and stirring speed 2 000 r·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.