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设计以温敏凝胶作为颗粒酶B质粒 (pGZMB) 和盐酸多柔比星 (DOX·HCl) 的共载载体以期实现基因和化疗药物联合治疗.选择支链型聚乙烯亚胺 (bPEI) 作为pGZMB的载体, 自组装形成bPEI/pGZMB纳米粒, 通过琼脂糖凝胶电泳和粒径电位结果研究形成bPEI/pGZMB纳米粒的最优质量比并对最优处方的外观形态、粒径分布、Zeta电位、转染能力进行考察.以胶凝温度为指标考察温敏凝胶基质普朗尼克F127与普朗尼克F68的用量及共载入bPEI/pGZMB纳米粒和DOX·HCl后对胶凝温度的影响, 结合胶凝时间和通针性结果优化共载bPEI/pGZMB纳米粒和DOX·HCl的F127/F68温敏凝胶 (DOX·HCl&bPEI/pGZMB-G) 处方, 并对其流变学、体外释放特性、凝胶材料的细胞毒性等进行评价.结果表明最优质量比90∶32制备的bPEI/pGZMB纳米粒形态圆整, 大小较为均一, 平均粒径为 (60.19±0.68) nm, 平均电位为 (8.86±1.08) m V, 且具有较高的转染效率.最优处方制备的DOX·HCl&bPEI/pGZMB-G在室温下为溶液状态, 当温度在37℃时可转变为凝胶状态, 流变学性质考察结果也说明DOX·HCl&bPEI/pGZMB-G具有良好的温度敏感特性并表现为假塑性流体特征;体外释放结果显示与溶液组相比温敏凝胶组具有一定的缓释特性;体外抗增殖实验证明了温敏凝胶材料细胞毒性较低, 安全性良好.以上结果表明以F127/F68为基质的温敏凝胶可作为DOX·HCl和pGZMB肿瘤治疗局部给药良好的共递送载体.“,”The objective of this study was to design pGZMB and DOX·HCl co-loaded thermo-sensitive hydrogel in order to achieve combination therapy of gene and chemotherapeutic drug.Branched polyethylenimine (bPEI) was used as the carrier of pGZMB to form bPEI/pGZMB nanoparticles by self-assembly.The optimal mass ratio of bPEI and pGZMB was studied by agarose gel electrophoresis, particle size and Zeta potential.The morphology of bPEI/pGZMB nanoparticles prepared by optimal prescription was observed via Transmission electron microscope (TEM).Particle size and Zeta potential of bPEI/pGZMB nanoparticles were measured by dynamic light scattering using a Malvern Zeta-Sizer Nano-ZS instrument.The transfection efficiency of bPEI/pGZMB nanoparticles was also conducted on Hep G2 cells.The dosage of pluronic F127 and pluronic F68 asthermo-sensitive hydrogel matrixes was investigated by gelation temperature.In addition, the influence of bPEI/pGZMB nanoparticles and DOX·HCl on the gelation temperature after co-loaded into hydrogel was also evaluated.The formulation of bPEI/pGZMB and DOX·HCl co-loaded F127/F68 thermo-sensitive hydrogel (DOX·HCl & bPEI/pGZMB-G) was optimized by gelation temperature, gelation time and inject ability.Moreover, physicochemical evaluation in terms of rheology, in vitro release characteristics as well as the cytotoxicity of thermosensitive hydrogel materials was also conducted.Rheological properties of thermo-sensitive hydrogel were performed by the method of vertebral lamina.In vitro release characteristics of DOX·HCl were studied by dialysis bag diffusion technique.The cytotoxicity of thermo-sensitive hydrogel materials was assessed via MTT assay.The results showed that bPEI/pGZMB nanoparticles prepared by the optimal mass ratio of 90∶ 32 exhibited spherical shape, mean particle size and Zeta potential of bPEI/pGZMB nanoparticles were (60.19 ±0.68) nm and (8.86 ± 1.08) m V, respectively.Besides, the transfection efficiency of bPEI/pGZMB nanoparticles was relatively high.DOX·HCl & bPEI/pGZMB-G prepared by the optimal prescription was in solution state at room temperature while it could turn into gel state when DOX·HCl & bPEI/pGZMB-G was maintained at a temperature of 37 °C.The rheology evaluation also indicated good temperature sensitive property of DOX·HCl & bPEI/pGZMB-G and appeared as pseudoplastic fluid.In vitro drug release exhibited that hydrogel groups had slower release characteristics compared with solution groups.Moreover, in vitro cytotoxicity study verified that the materials of thermo-sensitive hydrogel were safe enough.It was concluded that F127/F68 based thermo-sensitive hydrogel could serve as excellent co-delivery carrier of DOX·HCl and pGZMB for local cancer therapy.