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。
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.
作者
黄清昱
陈奇英
孙晟甲
吴帮卫
林杉
阿力木江·买买提江
HUANG Qingyu;CHEN Qiying;SUN Shengjia;WU Bangwei;LIN Shan;Alimujiang·MAIMAITIJIANG(Department of Cardiology,Huashan Hospital,Fudan University,Shanghai 200040,China)
出处
《基础医学与临床》
2024年第2期235-241,共7页
Basic and Clinical Medicine
关键词
血小板膜
利拉鲁肽
纳米颗粒
细胞功能
platelet membrane fragments
liraglutide
nanoparticles
cell functions
