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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。
Objective To prepare and evaluate mannan-modified chitosan coated lactic acid-glycolic acid copolymer (PLGA) nanoparticles to investigate their effects on macrophage cytotoxicity and macrophage uptake. Methods The ovalbumin (OVA) loaded PLGA nanoparticles were prepared by double emulsion method. The size and zeta potential of nanoparticles were determined by laser particle sizing and coating with mannose-modified chitosan. Transmission electron microscopy Appearance morphology, BCA method to measure the content of OVA drug loading and release. FITC-labeled OVA nanoparticles were co-incubated with macrophages (RAW 264.7), cell viability was measured by MTT assay, and the degree of uptake was assessed by fluorescence microscopy. Results The size and zeta potential of OVA-PLGA NPs increased with the increase of the concentration of chitosan coating solution (P <0.05). The drug load of OVA ranged from 7.2% to 8.4%. Chitosan and mannose-modified chitosan-coated FITC-OVA-PLGA nanoparticles co-incubated with RAW 264.7 had little effect on cell viability (P> 0.05), but could significantly promote the FITC-OVA-PLGA nanoparticles Granulocyte uptake (P <0.05). Conclusion The mannose-modified cationic nanoparticle system loaded with model antigens was preliminarily established, which provided a basis for the targeted study of antigen presenting cells in vivo.