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徐萍萍,杨利华,魏桂芝,张建祥,东亚君,梁志清.男性避孕纳米粒的制备与体外实验研究[J].陆军军医大学学报(原第三军医大学学报),2015,37(04):351-355.
Xu Pingping,Yang Lihua,Wei Guizhi,Zhang Jianxiang,Dong Yajun,Liang Zhiqing.Preparation and in vitro effect of FSHR32-44/PLGA nanoparticles as male contraceptive[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(04):351-355.
徐萍萍,杨利华,魏桂芝,张建祥,东亚君,梁志清.男性避孕纳米粒的制备与体外实验研究[J].陆军军医大学学报(原第三军医大学学报),2015,37(04):351-355. DOI:
Xu Pingping,Yang Lihua,Wei Guizhi,Zhang Jianxiang,Dong Yajun,Liang Zhiqing.Preparation and in vitro effect of FSHR32-44/PLGA nanoparticles as male contraceptive[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(04):351-355. DOI:
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(FSHR
32-44
)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(FSHR
32-44
)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)mV,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FSHR
32-44
避孕疫苗的载体。
Objective To synthesize nanoparticles (NP) with follicle-stimulating hormone receptor peptide (FSHR
32-44
) encapsulated by poly(lactic acid-glycolic acid) (PLGA)
which is biodegradable polymer material approved by Food and Drug Administration
and explore whether PLGA carrier is suitable for contraceptive vaccine. Methods Emulsification process was used for preparation of FSHR
32-44
-encapsulated PLGA NP (FSHR
32-44
/PLGA NP)
and the mo
rphology was observed with a transmission electron microscope. The particle size distribution and Zeta potential were detected with a laser potential particle size instrument
and the in vitro releasing characteristic of the NP was studied. A laser scanning confocal microscope was employed to observe the uptake capacity of bone marrow-derived dendritic cells (BMDCs) to soluble FSHR
32-44
and FSHR
32-44
/PLGA NP
and a flow cytometer was used to detect the phenotypic changes of BMDCs co-cultured with FSHR
32-44
PLGA NP and FSHR
32-44
/PLGA NP
respectively. Results FSHR
32-44
/PLGA NP had uniform particle size distribution and regular surface
and could slowly release antigens
with average particle size of 296.3±3.5 nm
Zeta potential of -16.1±0.4 mV
FSHR
32-44
encapsulation efficiency of 62.71%±2.83%
and total release rate of 98.3%. The in vitro cell test showed that FSHR
32-44
was easily up-taken by BMDCs in a dose- and time-dependent manner
and the expressions of CD40
CD86
CD83 and MHC-Ⅱ on the surface of BMDCs were up-regulated. Conclusion The FSHR
32-44
/PLGA NP have good encapsulation efficiency
and can be easily up-taken by BMDCs to promote maturation of BMDCs. PLGA can be used as the carrier of FSHR
32-44
contraceptive vaccine.
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