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目的制备防龋基因疫苗的DNA/PELA微球,并研究其在真核细胞中的表达情况。方法采用改进的溶剂挥发法双乳液(W1/O/W2)体系制备基因疫苗的DNA/PELA微球;测定了微球的形态大小、包封率及细胞毒性,将基因疫苗微球转染哺乳动物细胞,用流式细胞仪和RT-PCR测定在其中表达的情况。结果制备的DNA/PELA复合物多呈球状,平均粒径1.806μm,对细胞无毒性,可在体外转染哺乳动物细胞,并能正确地转录表达;结果证实基因疫苗微球转染效率低,提示微球中的DNA具有缓慢释放的特征。结论PELA可作为防龋基因疫苗的投递系统,是一种安全、有效的释放体系。
Objective To prepare DNA / PELA microspheres of anticaries caries gene vaccine and study its expression in eukaryotic cells. Methods DNA / PELA microspheres of gene vaccine were prepared by a modified solvent evaporation double emulsion (W1 / O / W2) system. Morphological size, entrapment efficiency and cytotoxicity of the microspheres were measured. Animal cells, in which the expression was measured by flow cytometry and RT-PCR. Results The DNA / PELA complexes were mostly spherical with an average particle diameter of 1.806μm. They were non-toxic to cells and transfected into mammalian cells in vitro and could be transcribed correctly. The results showed that the efficiency of transfection was low, Suggesting that DNA in the microspheres has a slow release profile. Conclusion PELA can be used as delivery system of anti-caries genetic vaccine, which is a safe and effective delivery system.