纳米乳剂的制备及特性鉴定

目的: 制备包裹BSA- FITC的纳米乳剂 (NEBSA -FITC), 研究其特性以及树突状细胞 (DC)和巨噬细胞对其摄取的效率。方法: 采用界面聚合法, 制备包裹BSA- FITC的纳米乳剂(NEBSA- FITC), 于电镜下观察其形态并测量其粒径。用凝胶层析法检测纳米乳剂的包裹率、载药量及稳定性。将小鼠骨髓DC及腹腔巨噬细胞与NEBSA -FITC共育后, 用流式细胞仪检测DC及腹腔巨噬细胞摄取NEBSA- FITC的阳性率。用激光共聚焦显微镜观察DC摄取NEBSA -FITC的能力。结果: 成功地制备出粒径为(25±10)nm的纳米乳剂, 其包裹率为 91%, 载药量为 0. 091g/L, 于 4℃放置 6个月后性质稳定。流式细胞仪检测显示, DC和腹腔巨噬细胞摄取NEBSA FITC的能力明显强于对游离BSA- FITC的摄取。激光共聚焦显微镜下观察到DC可摄取NEBSA -FITC。结论: 纳米乳剂具有良好的包裹率和载药量, 可有效地增强抗原递呈细胞对所载药物的摄取, 有望作为肿瘤抗原疫苗的新型载体。

Preparation and characterization of nanoemulsion

AIM: To prepare nanoemulsion-encapsulated BSA-FITC (NEBSA-FITC), study its characteristics, and measure its uptake by dendritic cells (DCs) and peritoneal macrophages. METHODS: NEBSA-FITC was prepared by a method of interfacial polymerization.The encapsulation rate, drug-carrying capacity and stability of the nanoemulsion were determined by Sephadex-G100 chromatography. The shape and size of NEBSA-FITC were observed under electron microscope. The uptake of NEBSA-FITC by DCs and macrophage cells was detected by FACS and laser confocal microscopy. RESULTS: The mean size of NEBSA-FITC was (25+/-10) nm. The encapsulation rate was 91%, the drug-carrying capacity was 0.091 g/L and NEBSA-FITC had a good stability. The FACS analysis showed that DCs and macrophage cells could take in more NEBSA-FITC than free BSA. The observation under laser confocal microscope found that NEBSA-FITC was located in the cytoplasm of DCs. CONCLUSION: Nanoemulsion can be efficiently taken by DCs and macrophage cells, and therefore may be promising efficient carrier of APCs-targeted antitumor vaccine.