Opanasopit Praneet, Apirakaramwong Auayporn, Ngawhirunpat Tanasait, Rojanarata Theerasak, Ruktanonchai Uracha
Nanotechnology for Drug/Gene Delivery Systems Group, Faculty of Pharmacy, Silpakorn University, Nakhon Pathom, Thailand.
AAPS PharmSciTech. 2008;9(1):67-74. doi: 10.1208/s12249-007-9007-7. Epub 2008 Jan 9.
The aim of this study was to investigate the possibility of using pectinate micro/nanoparticles as gene delivery systems. Pectinate micro/nanoparticles were produced by ionotropic gelation. Various factors were studied for their effects on the preparation of pectinate micro/nanoparticles: the pH of the pectin solution, the ratio of pectin to the cation, the concentration of pectin and the cation, and the type of cation (calcium ions, magnesium ions and manganese ions). After the preparation, the size and charge of the pectin micro/nanoparticles and their DNA incorporation efficiency were evaluated. The results showed that the particle sizes decreased with the decreased concentrations of pectin and cation. The type of cations affected the particle size. Sizes of calcium pectinate particles were larger than those of magnesium pectinate and manganese pectinate particles. The DNA loading efficiency showed that Ca-pectinate nanoparticles could entrap DNA up to 0.05 mg when the weight ratio of pectin:CaCl(2):DNA was 0.2:1:0.05. However, Mg-pectinate could entrap only 0.01 mg DNA when the weight ratio of pectin:MgCl(2):DNA was 1:100:0.01 The transfection efficiency of both Ca-pectinate and Mg-pectinate nanoparticles yielded relatively low levels of green fluorescent protein expression and low cytotoxicity in Huh7 cells. Given the negligible cytotoxic effects, these pectinate micro/nanoparticles can be considered as potential candidates for use as safe gene delivery carriers.
本研究的目的是探讨使用果胶微/纳米颗粒作为基因传递系统的可能性。果胶微/纳米颗粒通过离子凝胶法制备。研究了各种因素对果胶微/纳米颗粒制备的影响：果胶溶液的pH值、果胶与阳离子的比例、果胶和阳离子的浓度以及阳离子的类型（钙离子、镁离子和锰离子）。制备完成后，对果胶微/纳米颗粒的大小、电荷及其DNA包封效率进行了评估。结果表明，随着果胶和阳离子浓度的降低，颗粒尺寸减小。阳离子的类型影响颗粒大小。果胶钙颗粒的尺寸大于果胶镁颗粒和果胶锰颗粒。DNA负载效率表明，当果胶:氯化钙:DNA的重量比为0.2:1:0.05时，果胶钙纳米颗粒可包封高达0.05 mg的DNA。然而，当果胶:氯化镁:DNA的重量比为1:100:0.01时，果胶镁只能包封0.01 mg的DNA。果胶钙和果胶镁纳米颗粒的转染效率在Huh7细胞中产生的绿色荧光蛋白表达水平相对较低且细胞毒性较低。鉴于细胞毒性作用可忽略不计，这些果胶微/纳米颗粒可被视为用作安全基因传递载体的潜在候选物。
