IN VITRO EVALUATION OF INTERLEUKIN-10 GENE DELIVERY INTO DORSAL ROOT GANGLION CELLS MEDIATED BY PEI-g-MPEG
PEI-g-MPEG/白细胞介素-10基因传递系统转染背根神经节细胞的体外性能研究

The use of non-viral carrier,polyethyleneimine-g-methoxypoly(ethylene glycol) (PEI-g-MPEG) for interleukin-10 (IL-10) gene delivery into rat dorsal root ganglion cells (DRGs) was examined in vitro.Electrophoresis,ethidium bromide exclusion,SEM,size and zeta (ζ) potential measurements were performed to characterize the nanocomplex,PEI-g-MPEG/pDNA.In addition,the cytotoxicity,transfection efficiency and IL-10 protein expression level in comparison with lipofectamine as a delivery agent were determined.Results showed that plasmid DNA was completely complexed by PEI-g-MPEG at a nitrogen-to-phosphor (N/P) ratio of 5.The PEI-g-MPEG/pDNA complexes were well-distributed and spherical with smooth surface.As the proportion of PEI-g-MPEG increased,i.e.when N/P ratio increased from 1 to 30,the size of the PEI-g-MPEG/DNA complexes decreased from 229 nm to 130 nm,while the surface charge increased from -19 mV to 32 mV,which led to the increased cytotoxicity.Flow cytometry revealed that the highest transfection efficiency (24%) using PEI-g-MPEG as a delivery agent was obtained at N/P charge ratio of 15,which was higher than that of lipofectamine (17%) at the same N/P value.After transfection,rat IL-10 expression in DRGs primary culture assessed using ELISA was observed to localize in the cytoplasm at 48 h in concordance with the result obtained for EGFP expression.These results indicated that PEI-g-MPEG could be a potential candidate for gene delivery during the therapy of neuropathic pain.