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

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.