甘露糖修饰壳聚糖对巨噬细胞摄取纳米粒的影响

目的制备和评价甘露糖修饰壳聚糖包衣乳酸-羟基乙酸共聚物（PLGA）纳米粒,考察其对巨噬细胞毒性和对巨噬摄取功能的影响。方法采用复乳法制备负载卵清蛋白（OVA）的PLGA纳米粒,并经甘露糖修饰壳聚糖包衣处理后用激光粒度测定仪测定该纳米粒的大小与ζ电位,透射电镜观察纳米粒的外观形态,BCA法测定OVA含量后计算载药量与释放度。负载异硫氰酸荧光素（FITC）标记的OVA纳米粒与巨噬细胞（RAW 264.7）共孵育,MTT法测定细胞存活率,荧光显微镜考察摄取程度。结果 OVA-PLGA纳米粒的大小和ζ电位均随壳聚糖包衣液浓度的增加而变大（P〈0.05）,OVA载药量范围为7.2%～8.4%。壳聚糖与甘露糖修饰壳聚糖包衣FITC-OVA-PLGA纳米粒与RAW 264.7共孵育后,对细胞存活率的影响不大（P〉0.05）,但可明显促进FITC-OVA-PLGA纳米粒的巨噬细胞摄取（P〈0.05）。结论初步建立了负载模型抗原的甘露糖修饰阳离子型纳米粒系统,这为体内抗原递呈细胞靶向性研究提供了依据。

Effects of Mannose Modified Chitosan on the Macrophage Uptake of Nanoparticles

Objective To prepare and evaluate mannose modified chitosan（Man-CS） coated poly lactic-co-glycolic acid（PLGA） nanoparticles（NPs）,observe its cytotocity and effects on the uptake of macrophages.Method Ovalbumin（OVA） loaded PLGA NPs were prepared by double emulsion method and coated with CS or Man-CS.The size and zeta potentional of NPs were detected by a laser particle size analyzer,BCA method was used to measure OVA concentration and calculate the drug loading and release rate.The cytotocixty and phagocytosis of FITC-OVA loaded NPs were investigated by MTT method and fluorescence microscope after incubating with macrophage cells（RAW 264.7）.Results Both of the particle sizes and zeta potentionals of the OVA-PLGA nanoparticles increased with increasing the concentration of CS,and the loading amount of OVA was in the range of 7.2%～8.4%.No significant cytotoxicity was observed after incubating CS or Man-CS coating FITC-OVA-PLGA nanoparticles with RAW 264.7（P0.05） but noticeable phagocytosis effects were obtained（P0.05）.Conclusions The model antigen loaded cationic nanoparticles modified with mannose-chitosan are prepared and it will provide evidences for the rsearch of antigen presentation cell targeting delivery systems.