文章摘要
隆 梅,张 钟,张 攀,陆 琴,方 超.巨噬细胞膜伪装纳米颗粒的制备和功能表征[J].,2021,(14):2601-2606
巨噬细胞膜伪装纳米颗粒的制备和功能表征
Preparation and Functional Characterization of Macrophage Membrane Camouflaged Nanoparticles
投稿时间:2020-12-06  修订日期:2020-12-28
DOI:10.13241/j.cnki.pmb.2021.14.001
中文关键词: 巨噬细胞  细胞膜  纳米颗粒  阿霉素  药物递送
英文关键词: Macrophages  Cell membrane  Nanoparticles  Doxorubicin  Drug delivery
基金项目:国家自然科学基金项目(81773274;81572998)
作者单位E-mail
隆 梅 上海交通大学医学院药理学与化学生物学系 上海 200025 maysitu@163.com 
张 钟 上海交通大学医学院药理学与化学生物学系 上海 200025  
张 攀 上海交通大学医学院药理学与化学生物学系 上海 200025  
陆 琴 上海交通大学医学院药理学与化学生物学系 上海 200025  
方 超 上海交通大学医学院药理学与化学生物学系 上海 200025  
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中文摘要:
      摘要 目的:巨噬细胞具有炎症趋化能力,近年来巨噬细胞膜伪装的纳米递送载体引起研究者的广泛关注。本文提供了一种巨噬细胞膜伪装纳米颗粒的方法,即摄取-挤出法,并对该法制得的纳米颗粒进行表征,考察纳米颗粒在不同细胞中的摄取。方法:利用溶胶-凝胶法制备装载阿霉素的介孔硅(DMSN)纳米颗粒,再利用RAW 264.7巨噬细胞吞噬DMSN,最后将巨噬细胞连续挤出制得巨噬细胞膜伪装的载有阿霉素的介孔硅(DMSN@CM)纳米颗粒。动态光散射激光粒度仪(DLS)测定DMSN@CM颗粒的粒径和表面电位,透射电子显微镜(TEM)观察纳米颗粒形态,聚丙烯酰胺凝胶电泳(SDS-PAGE)验证细胞膜的成功伪装。然后通过激光共聚焦显微镜与流式细胞术共同考察了DMSN@CM在不同细胞中的摄取情况。结果:成功制备了DMSN和DMSN@CM纳米颗粒。DMSN粒径为116.7±3.2 nm,zeta表面电势为 -29.5± 1.3 mV;MSN@CM粒径为128.0±9.3 nm,zeta表面电势为 -26.7 ±1.2 mV。TEM与SDS-PAGE共同验证了DMSN@CM表面细胞膜的成功包覆。细胞摄取试验表明巨噬细胞膜的伪装可以抑制RAW 264.7细胞对DMSN@CM的摄取;促进MDA-MB-231细胞对DMSN@CM的摄取。结论:利用摄取-挤出法成功构建了DMSN@CM纳米颗粒,该法简便高效,为纳米颗粒的细胞膜伪装提供了一种新的手段。
英文摘要:
      ABSTRACT Objective: Macrophage is a kind of immune cells with the ability of inflammatory chemotaxis. In recent years, macrophage membrane-camouflaged nanoparticle delivery system has attracted wide attention of researchers. Herein, a new method for macrophage membrane to camouflage nanoparticle-uptake-extrusion method is developed, and then we characterize the nanoparticles prepared by this method, and investigate the uptake of nanoparticles in different cells. Methods: DMSN nanoparticles loaded with doxorubicin were prepared by sol-gel method, and then DMSN was phagocytized by RAW 264.7 macrophages. Finally, macrophages were continuously extruded to prepare DMSN@CM nanoparticles. The particle size and zeta potential of DMSN@CM were measured by nanoparticle size analyzer. The morphology of nanoparticles was observed by transmission electron microscope. Polyacrylamide gel electrophoresis was used to verify the successful camouflage of cell membrane. Then the uptake of DMSN@CM in different cells was investigated by laser confocal microscope and flow cytometry. Results: DMSN and DMSN@CM nanoparticles were successfully prepared. The size of DMSN was 116.7±3.2 nm, and its zeta potential was -29.5±1.3 mV; The size of MSN@CM was 128.0±9.3 nm, and its zeta potential was -26.7±1.2 mV. TEM and SDS-PAGE jointly verified the successful coating of cell membrane. Cell uptake assay showed that the camouflage of macrophage membrane could inhibit the uptake of DMSN@CM in RAW 264.7 cells, and promote the uptake of DMSN@CM in MDA-MB-231 cells. Conclusion: Thus, we provide a simple and efficient method for macrophage membrane to camouflage nanoparticles-uptake-extrusion method, and successful constructed DMSN@CM nanoparticles based on this method.
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