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以可生物降解材料聚乳酸聚乙醇酸共聚物(Poly-dl-lactic-co-glycolic,PLGA)为原料,采用多相乳化技术制备载VEGF纳米粒子。并对纳米粒子的粒径,VEGF含量,体外释放等进行了测定。VEGF纳米粒子和VEGF裸质粒被注射到兔下肢缺血模型的缺血部位,通过RT-PCR,免疫组化和血管造影等技术来验证基因治疗的效果,评价VEGF纳米粒子作为基因载体在动物模型基因治疗中的效率。制备的VEGF纳米粒子的平均粒径约为300nm,包埋效率在96%以上,纳米粒子中VEGF含量约4%。可在体外维持恒定释放约两周。两周基因注射结果表明VEGF-NP治疗组与裸质粒VEGF治疗组的毛细血管密度明显高于对照组,VEGF纳米粒子组(81.22permm2),对照组(29.54mm2),两者有显著性差异(P<0.05)。RT-PCR结果显示VEGF纳米粒子组表达(31.79au*mm)明显高于VEGF裸质粒组(9.15au*mm)。在动物模型中VEGF纳米粒子是比裸质粒DNA更好的基因载体系统,结果显示了纳米粒子可望在人类基因治疗中得到很好的应用。
Poly (dl-lactic-co-glycolic) (PLGA), a biodegradable material, was used as a raw material to prepare VEGF-loaded nanoparticle. The particle size, VEGF content and in vitro release of nanoparticles were also determined. VEGF nanoparticles and VEGF naked plasmid were injected into the ischemic region of the rabbit lower limb ischemia model to verify the effect of gene therapy by RT-PCR, immunohistochemistry and angiography and to evaluate the effect of VEGF nanoparticle as a gene carrier in animal model Gene therapy efficiency. The average diameter of the prepared VEGF nanoparticles was about 300 nm, the embedding efficiency was above 96%, and the content of VEGF in the nanoparticles was about 4%. A constant release can be maintained in vitro for about two weeks. The two-week gene injection results showed that the capillary density of VEGF-NP-treated group and naked plasmid VEGF-treated group was significantly higher than that of the control group, the VEGF nanoparticle group (81.22permm2) and the control group (29.54mm2) P <0.05). The results of RT-PCR showed that the expression of VEGF nanoparticles (31.79au * mm) was significantly higher than that of VEGF naked plasmid (9.15au * mm). In animal models, VEGF nanoparticles are better gene delivery systems than naked plasmid DNA. The results show that nanoparticles are expected to be well-suited for human gene therapy.