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载GOD/Fe3O4的PLGA纳米粒子对视网膜母细胞瘤疗效的体外研究
作者:全鲁亚 乔斌 王志刚 张屿 杜之渝 
单位:400010  重庆 重庆医科大学附属第二医院眼科(全鲁亚、杜之渝) 400010  重庆 重庆医科大学超声分子影像重庆市重点实验室(全鲁亚、乔斌、王志刚) 400010  重庆 重庆明达眼科医院眼科(张屿) 
关键词:视网膜母细胞瘤 葡萄糖氧化酶 PLGA纳米粒子 超顺磁性四氧化三铁纳米粒子 
分类号:R739.7+2
出版年,卷(期):页码:2019,44(5):382-387
摘要:

[摘要]  目的  探讨一种新型的包裹葡萄糖氧化酶(GOD)/超顺磁性四氧化三铁纳米粒(Fe3O4Nps)的聚乳酸-羟基乙酸共聚物(PLGA)纳米粒子对视网膜母细胞瘤(RB)体外治疗的效果。方法  利用双乳化法制备包裹GOD/Fe3O4PLGA纳米粒子,对其粒径大小、电位、外部形态和内部结构等相关表征进行检测。依据包含的不同纳米粒子设立空白对照组、Fe3O4/PLGA(PF)组、GOD/PLGA(PG)组、GOD/Fe3O4/PLGA(PFG)组;设立不同浓度(00.06250.1250.25 μg/ml)PFG纳米粒子组,以验证其毒性作用。检测不同纳米粒子组与Y79细胞共孵育后对Y79细胞的毒性作用、存活情况及活性氧产生情况。结果  成功制备出包裹了GOD/Fe3O4PLGA纳米粒子,其平均粒径为299.3 nm, 形态均一呈圆形。纳米粒子与Y79细胞共孵育后被Y79细胞吞噬,产生大量活性氧;细胞毒性实验结果显示,PFG Y79细胞存活率[(53.648±2.565)%]明显低于空白对照组[(100.028±4.491)%]PF[(97.782±17.520)%]PG[(87.438±3.537)%] (F=21.226P<0.01)0.25 μg/mlPFG纳米粒子组Y79细胞存活率[(51.770±1.529)%]明显低于对照组[(100.000±5.021)%]0.0625 μg/ml[(85.723±6.903)%]0.125 μg/ml[(74.535±8.282)%](F=34.593P<0.01)。激光共聚焦显微镜下观察,结果显示PFGY79细胞大量死亡。结论  成功制备了包裹GOD/Fe3O4的新型PLGA纳米粒子;该纳米粒子具有良好的活性氧产生能力,能杀死Y79肿瘤细胞,有望成为一种治疗RB的新方法。

 [Abstract]  Objective  To evaluate the therapeutic potential of a novel type of Poly (lactic-co-glycolic acid) (PLGA) nanoparticles loaded with glucose oxidase (GOD)/superparamagnetic iron oxide nanoparticles (Fe3O4Nps) on retinoblastoma (RB) cells in vitro. Methods  PLGA nanoparticles loaded with GOD/Fe3O4 (PFG) were prepared by double emulsification. Their particle size, potential, external morphology, and internal structure were examined. Particles that made of PLGA (control), PLGA loaded with GOD (PG), and PLGA loaded with Fe3O4 (PF) are served as control. Y79 cells that were incubated with different particles are termed control group, PF group, PG group, and PFG group. After co-incubation with nanoparticles, cell viability, and reactive oxygen species production were detected. Results  PLGA nanoparticles loaded with GOD/Fe3O4 were successfully prepared. The form of PLGA nanoparticles was uniform and showed a round shape with a diameter of 299.3 nm. The nanoparticles were engulfed by Y79 cells after co-incubation with Y79 cells, producing a large number of reactive oxygen species. Cytotoxicity test results showed that the cell viability of Y79 cells in PLGA nanoparticle group coated with GOD/Fe3O4 [(53.648±2.565)%] was significantly lower than that in control group [(100.028±4.491)%], PF group [(97.782±17.520)%] or PG group [(87.438±3.537)%](F=21.226, P<0.01); The cell viability of Y79 cells in 0.25 μg/ml PFG nanoparticle group [(51.770±1.529)%] was significantly lower than that in control group [(100.000±5.021)%], 0.0625 μg/ml group [(85.723±6.903)%] and 0.125 μg/ml group [(74.535±8.282)%] (F=34.593, P<0.05). Massive cell death was detected in the PFG group under laser confocal microscope. Conclusions  The novel type of PLGA nanoparticles loaded with GOD/ Fe3O4 toxic to Y79 tumor cells with a good reactive oxygen generation ability. It provides a potential treatment for RB.

基金项目:
重庆市科学技术委员会基础与前沿项目(cstc2018jcyjAX0562)
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