氧化微环境中普鲁士蓝纳米酶促进MC3T3-E1细胞成骨分化  

Prussian blue nanozyme promotes osteogenic differentiation of MC3T3⁃E1 cells in an oxidative microenvironment

作　　者：陈露 唐诗佳 钟彩英 胡克 章非敏[1,2,3] CHEN Lu;TANG Shijia;ZHONG Caiying;HU Ke;ZHANG Feimin(Department of Prosthodontics,the Affiliated Stomatological Hospital of Nanjing Medical University,Nanjing 210029;Jiangsu Province Key Laboratory of Oral Diseases,Nanjing 210029;Jiangsu Province Engineering Research Center of Stomatological Translational Medicine,Nanjing 210029;Key Laboratory of Clinical Engineering,School of Biomedical Engineering and Informatics,Nanjing Medical University,Nanjing 211166,China)

机构地区：[1]南京医科大学附属口腔医院口腔修复科,江苏南京210029 [2]南京医科大学附属口腔医院,江苏省口腔疾病研究重点实验室,江苏南京210029 [3]南京医科大学附属口腔医院,江苏省口腔转化医学工程研究中心,江苏南京210029 [4]南京医科大学生物医学工程与信息学院,临床医学工程校重点实验室,江苏南京211166

出　　处：《南京医科大学学报（自然科学版）》2023年第5期669-677,共9页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家重点研发计划纳米科技重点专项(2021YFA1201302/2021YFA1201300);江苏高校优势学科建设工程资助项目(2018-87)。

摘　　要：目的:探索氧化微环境中普鲁士蓝纳米酶对MC3T3-E1细胞成骨分化的影响。方法:CCK-8法和流式细胞仪检测不同浓度普鲁士蓝纳米酶的生物相容性;使用二甲酚橙法、WST-8法检测不同浓度普鲁士蓝纳米酶的模拟酶活性,DCFH-DA探针免疫荧光法检测普鲁士蓝纳米酶对细胞内活性氧的清除能力;通过碱性磷酸酶(alkaline phosphatase,ALP)染色及活性检测、茜素红染色、实时荧光定量逆转录PCR评估其氧化微环境下对MC3T3-E1细胞成骨分化的影响。结果:CCK-8和流式凋亡结果显示,0~100μg/mL普鲁士蓝纳米酶具有良好的生物相容性;二甲酚橙法、WST-8法结果显示,普鲁士蓝具有良好的模拟酶活性并且呈时间和剂量依赖性;DCFH-DA探针免疫荧光结果显示,普鲁士蓝纳米酶对细胞内活性氧也有较好的清除能力;ALP染色及活性检测、茜素红染色、实时荧光定量逆转录PCR结果显示,氧化微环境中普鲁士蓝纳米酶对MC3T3-E1细胞的成骨分化能力具有显著增强作用。结论:普鲁士蓝纳米酶可以增强MC3T3-E1细胞的抗氧化活性,促进氧化环境中细胞的成骨分化能力。Objective:This study aims to explore the effect of Prussian blue nanozyme on osteogenic differentiation of MC3T3⁃E1 cells in an oxidative microenvironment.Methods:Biocompatibility of different concentrations of Prussian blue nanozymes was detected by CCK⁃8 assay and flow cytometry;mimetic enzyme activity of Prussian blue nanozymes at different concentrations was measured by xylenol orange assay and WST⁃8 assay,and intracellular reactive oxygen species scavenging activity of Prussian blue nanozymes was detected by DCFH⁃DA probe immunofluorescence assay;and the osteogenesis differentiation of MC3T3⁃E1 cells under oxidative microenvironment was evaluated by alkaline phosphatase(ALP)staining and activity assay,alizarin red staining and real⁃time fluorescence quantitative reverse transcription PCR.Results:CCK⁃8 and flow cytometry showed good biocompatibility of Prussian blue nanozyme between 0⁃100μg/mL concentrations;xylenol orange assay and WST⁃8 assay showed good mimetic enzymatic activity of Prussian blue nanozyme in a concentration and time⁃dependent manner;DCFH⁃DA probe immunofluorescence showed that Prussian blue nanozyme also had good ability to scavenge intracellular reactive oxygen species;ALP staining and activity assay,alizarin red staining and real⁃time fluorescence quantitative reverse transcription PCR showed that Prussian blue nanozyme can significantly enhance osteogenesis differentiation of MC3T3⁃E1 cells in oxidative microenvironment.Conclusion:Prussian blue nanozyme can enhance the antioxidant activity of MC3T3⁃E1 cells and promotes the osteogenic differentiation of cells in an oxidative environment.

关 键 词：普鲁士蓝纳米酶 活性氧 抗氧化 成骨分化 骨组织工程 

分 类 号：R318.08[医药卫生—生物医学工程]