基础医学与临床 ›› 2024, Vol. 44 ›› Issue (2): 235-241.doi: 10.16352/j.issn.1001-6325.2024.02.0235

• 研究论文 • 上一篇    下一篇

Lir@BSA-PMF纳米颗粒的制备及其细胞功能验证

黄清昱, 陈奇英, 孙晟甲, 吴帮卫, 林杉, 阿力木江·买买提江*   

  1. 复旦大学附属华山医院 心内科,上海 200040
  • 收稿日期:2023-09-06 修回日期:2023-11-29 发布日期:2024-02-05
  • 通讯作者: *alimujiang2020@163.com

Preparation of Lir@BSA-PMF nanoparticles and verification of their cell functions

HUANG Qingyu, CHEN Qiying, SUN Shengjia, WU Bangwei, LIN Shan, Alimujiang·MAIMAITIJIANG*   

  1. Department of Cardiology, Huashan Hospital, Fudan University, Shanghai 200040, China
  • Received:2023-09-06 Revised:2023-11-29 Published:2024-02-05
  • Contact: *alimujiang2020@163.com

摘要: 目的 利用生物膜覆盖纳米粒子的“bottom-up”表面纳米工程化学技术合成牛血清白蛋白(BSA)负载利拉鲁肽(Lir)包裹血小板膜碎片(PMF)的纳米颗粒,并验证其细胞相容性以及抗氧化应激损伤的能力。方法 按照既往文献中的方法提取PMF;利用自组装法制备Lir@BSA纳米颗粒;通过共挤压的方式将PMF包覆在Lir@BSA纳米颗粒表面制备Lir@BSA-PMF。利用粒径、电位、透射电镜、粒径稳定性表征Lir@BSA-PMF颗粒的理化指标;利用酶联免疫吸附试验法计算利拉鲁肽的包封率、负载率和累计释放率;进一步利用SDS-PAGE分析Lir@BSA-PMF仿生纳米载体上是否存在血小板膜完整的膜蛋白质结构;同时利用CCK-8法验证材料的生物相容性;活性氧(ROS)实验探究了Lir@BSA-PMF仿生纳米载体对细胞氧化损伤的影响;最后通过细胞吞噬实验验证细胞对Lir@BSA-PMF仿生纳米载体的摄取效果。结果 制备的Lir@BSA-PMF的纳米粒子粒径可稳定在25 nm,形貌为球形,Zeta电位值为-25.5 mV。利拉鲁肽的包封率、负载率和累计释放率分别为85.56%、7.96%和77.06%。SDS-PAGE分析结果显示Lir@BSA-PMF仿生纳米载体上保留了血小板膜完整的膜蛋白质结构。CCK-8法验证纳米材料无细胞毒性;ROS结果显示Lir@BSA-PMF纳米材料具有明显的抗氧化性。细胞吞噬结果表明,细胞对Lir@BSA-PMF纳米颗粒的吞噬效果良好。结论 Lir@BSA-PMF复合纳米粒子合成成功,体外对细胞存活无影响,可被细胞摄取,并具有明显抗氧化损伤能力。

关键词: 血小板膜, 利拉鲁肽, 纳米颗粒, 细胞功能

Abstract: Objective To synthesize bovine serum albumin(BSA)-loaded liraqlutide (Lir)-nanoparticles coated with platelet membrane fragments (PMF) using a“bottom-up” nano-engineering chemistry technique, and to evaluate their cyto-compatibility and potential function of anti-oxidative stress. Methods PMF was extracted as reported previously. Lir@BSA nanoparticles were prepared by self-assembly method. PMF was coated on the surface of Lir@BSA nanoparticles by co-extrusion to prepare Lir@BSA-PMF. The physical and chemical properties of Lir@BSA-PMF particles were characterized as particle size, Zeta potential, transmission electron microscopy and particle size stability. The encapsulation efficiency, loading efficiency and cumulative release efficiency of liraglutide were calculated by enzyme-linked immunosorbent assay. Further, SDS-PAGE was used to analyze whether there was a similar membrane protein distribution of platelet membrane on Lir@BSA-PMF bionicnanocarrier.CCK-8 assay was used to verify the biocompatibility of the materials. Reactive oxygen species (ROS) experiment was used to explore the effect of Lir@BSA-PMF on cell oxidative damage. The uptake of cells on Lir@BSA-PMF bionic nano capsules was verified by cell phagocytosis experiment. Results Lir@BSA-PMF nanoparticles had a stable particle size of 25 nm with a spherical morphology, and a Zeta potential value of -25.5 mV. The encapsulation efficiency, loading efficiency and cumulative release efficiency of liraglutide were 85.56%, 7.96% and 77.06%, respectively. SDS-PAGE analysis showed that the Lir@BSA-PMF bio-mimetic nano capsules retained the similar membrane protein distribution as platelet membrane. CCK-8 assay verified that the nanomaterials were non-cytotoxic. ROS results showed that Lir@BSA-PMF nanomaterials had obvious antioxidant properties. The results of cell phagocytosis showed that the cells had a good phagocytosis effect on Lir@BSA-PMF nanoparticles. Conclusions The nanoparticles Lir@BSA-PMF are successfully synthesized and have no effects on cells viability in vitro. The particles are taken up by cells and show a significant function of antioxidant damage.

Key words: platelet membrane fragments, liraglutide, nanoparticles, cell functions

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