四氧化三锰纳米颗粒抗氧化损伤保护骨髓间充质干细胞的伸展功能  

Protection of manganese oxide nanoparticles for bone marrow mesenchymal stem cell spreading against oxidative stress

作　　者：田沁玉 田兴贵 田壮 眭翔 刘舒云 鲁晓波 郭全义 Tian Qinyu;Tian Xinggui;Tian Zhuang;Sui Xiang;Liu Shuyun;Lu Xiaobo;Guo Quanyi(Department of Bone and Joint Surgery,Affiliated Hospital of Southwest Medical University,Luzhou 646000,Sichuan Province,China;Institute of Orthopedics,First Medical Center,Chinese PLA General Hospital,Beijing Key Lab of Regenerative Medicine in Orthopedics,Key Laboratory of Musculoskeletal Trauma&War Injuries,PLA,Beijing 100853,China;University Center of Orthopaedic,Trauma and Plastic Surgery,University Hospital Carl Gustav Carus at Technische Universität Dresden,Dresden 01307,Germany;Ninth Clinical School of Peking University,Beijing Shijitan Hospital,Beijing 100038,China)

机构地区：[1]西南医科大学附属医院骨与关节外科,四川省泸州市646000 [2]中国人民解放军总医院第一医学中心骨科研究所,骨科再生医学北京市重点实验室,全军骨科战创伤重点实验室,北京市100853 [3]德累斯顿工业大学附属大学医院骨科,德国德累斯顿01307 [4]北京大学第九临床医学院,北京世纪坛医院,北京市100038

出　　处：《中国组织工程研究》2023年第6期821-826,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家重点研发计划(2019YFA0110600),项目负责人:郭全义。

摘　　要：背景:由于干细胞移植微环境原位产生的大量自由基造成移植干细胞伸展功能受损,进而降低细胞的移植存活率。近10年来,大量抗氧化应激纳米酶研究的出现为保护干细胞伸展功能提供了新的方法。四氧化三锰(Mn_(3)O_(4))作为一种新型纳米酶,含有的锰作为人体微量元素之一,具有抗氧化应激性能以及生物可降解性,可作为一种保护干细胞伸展功能的新型纳米材料。目的:制备纳米酶四氧化三锰颗粒,检测其在氧化应激环境中对骨髓间充质干细胞抗氧化作用和伸展功能的影响。方法:水热法制备四氧化三锰纳米颗粒,用扫描电子显微镜、X射线衍射分别表征材料形貌和结构;动态光散射仪检测模拟人体环境下的颗粒粒径和Zeta电位;在中性环境下检测四氧化三锰的抗氧化能力,使用CCK-8和活-死染色法检测四氧化三锰对骨髓间充质干细胞的生物毒性;过氧化氢诱导骨髓间充质干细胞氧化应激,检测四氧化三锰对骨髓间充质干细胞在氧化应激状态下抗氧化能力及伸展能力的影响。结果与结论:①扫描电子显微镜和X射线衍射测试结果显示该材料为平均直径70-80 nm的四氧化三锰纳米颗粒;动态光散射仪检测显示,在模拟人体缓冲环境下的四氧化三锰的粒径约为100 nm,Zeta电位约为+20 mV,抗氧化性能实验显示四氧化三锰具有一定的抗氧化能力;②CCK-8、活-死染色、活性氧清除实验显示四氧化三锰在40 mg/L的质量浓度下对骨髓间充质干细胞无生物毒性且有一定的活性氧清除能力;③细胞伸展实验显示过氧化氢对骨髓间充质干细胞的伸展功能具有明显抑制作用,而四氧化三锰加入细胞培养微环境后,细胞伸展面积较对照组无明显统计学差异(P>0.05),且单独使用四氧化三锰不会抑制骨髓间充质干细胞的伸展功能(P>0.05);④结果表明,在细胞微环境中加入四氧化三锰纳米颗粒能够抵抗氧化应BACKGROUND:During stem cell transplantation,the generation of a large number of free radicals damages the cells in situ,and then reducing the cell survival.In the past decade,the emergence of nanozymes provides a new method for scavenging excess free radicals.Among them,manganese oxide(Mn_(3)O_(4))is one of the promising nanomaterials,because it could provide Mn trace elements in human body as well as its well antioxidative stress properties and biodegradability.Mn_(3)O_(4)can be used as a new type of nanomaterial to protect the extension function of stem cells.OBJECTIVE:To produce Mn_(3)O_(4)nanoparticles for detecting its effect on scavenging and spreading function of bone marrow mesenchymal stem cells during oxidative stress.METHODS:Mn_(3)O_(4)nanoparticles were prepared by hydrothermal method.Scanning electron microscope and X-ray diffraction were used to characterize the morphology and structure of Mn_(3)O_(4),respectively.The particle size and zetapotential data of the nanoparticle in human-simulated environment were detected by the dynamic light scattering.The antioxidant capacity of Mn_(3)O_(4)was tested in neutral environment.The CCK-8 and live-dead staining assays were utilized to verify the biological toxicity of Mn_(3)O_(4)to bone marrow mesenchymal stem cells.H_(2)O_(2)was used to induce oxidative stress of bone marrow mesenchymal stem cells.The effect of Mn_(3)O_(4)on antioxidant ability and spreading ability of bone marrow mesenchymal stem cells was detected under oxidative stress.RESULTS AND CONCLUSION:(1)The results of scanning electron microscope and X-ray diffraction exhibited that the material was Mn_(3)O_(4)with average particle size about 70-80 nm.The dynamic light scattering showed that the particle size of the nanoparticles in PBS solution was about 100 nm,and the Zeta potential was about+20 mV.Antioxidant performance experiments showed that Mn_(3)O_(4)had a certain antioxidant capacity.(2)The CCK-8 assay,live-death staining and reactive oxygen scavenging experiment demonstrated that Mn_(3

关 键 词：四氧化三锰 纳米颗粒 纳米酶 抗氧化应激 活性氧清除 干细胞移植 干细胞伸展 干细胞保护 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学] R318.08