男性避孕纳米粒的制备与体外实验研究

发布时间：2018-03-02 04:04

  本文关键词： FSHR PLGA 纳米疫苗 避孕　出处：《第三军医大学学报》2015年04期 　论文类型：期刊论文

【摘要】：目的构建聚乳酸-羟乙酸共聚物(polylactic acid-glycolic acid,PLGA)包载促卵泡生成素受体表位肽(FSHR32-44)的纳米粒,探讨PLGA材料作为避孕疫苗载体的可行性。方法使用PLGA为载体,制备出包载促卵泡生成素受体的B细胞表位(FSHR32-44)的纳米粒,透射电镜(TEM)观察表观形态,粒径仪检测粒径分布范围和Zeta电位,体外释放试验检测其理化特性。激光共聚焦显微镜观察小鼠骨髓来源树突状细胞(DCs)吞噬FSHR可溶性多肽和FSHR/PLGA NP的能力,流式细胞仪检测FSHR、PLGA NP和FSHR/PLGA NP促骨髓来源DCs成熟能力。结果单乳-溶剂挥发法制备的FSHR/PLGA NP大小粒径分布均匀、表面规整,粒径(296.3±3.5)nm,Zeta电位(-16.1±0.4)m V,FSHR包封率(62.71±2.83)%,并具有缓慢释放抗原的能力;体外细胞水平实验结果显示FSHR表位肽纳米粒易于被DCs摄取,有剂量和时间依赖性,能显著上调DCs表面CD40、CD86、CD83和MHC-Ⅱ功能分子的表达。结论 PLGA包载FSHR多肽有良好的包封率,易被DCs吞噬,并有强的促进DCs成熟作用,可以作为FSHR32-44避孕疫苗的载体。
[Abstract]:Objective to construct the nanoparticles of poly (lactic acid) -glycolactic acid-glycolic acidoga (PLGA) coated with follicle stimulating hormone (FSHR32-44), and to explore the feasibility of using PLGA as a contraceptive vaccine carrier. Methods PLGA was used as the carrier. The B cell epitope (FSHR32-44) loaded with follicle stimulating hormone receptor (FSHR32-44) was prepared. The apparent morphology was observed by transmission electron microscopy (TEM). The particle size distribution and Zeta potential were measured by particle size analyzer. Laser confocal microscopy was used to observe the ability of murine bone marrow-derived dendritic cells to phagocytosis of FSHR soluble polypeptides and FSHR/PLGA NP. Results the size distribution of FSHR/PLGA NP prepared by single emulsion solvent volatilization method was uniform, the surface of FSHR/PLGA NP was regular, the encapsulation efficiency of FSHR/PLGA NP was 62.71 卤2.83%, and the encapsulation efficiency of FSHR/PLGA NP was 62.71 卤2.83%. The results of cell level assay in vitro showed that FSHR epitope peptide nanoparticles were easy to be ingested by DCs in a dose-and time-dependent manner, and could significantly up-regulate the expression of CD40, CD86, CD83 and MHC- 鈪，

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