Page 87 - 南京医科大学学报自然科学版

第43卷第5期 南京医科大学学报（自然科学版）
2023年5月 Journal of Nanjing Medical University（Natural Sciences） ·669 ·

·基础研究·

氧化微环境中普鲁士蓝纳米酶促进MC3T3⁃E1细胞成骨分化

陈 露 1，2，3 ，唐诗佳 1，2，3 ，钟彩英 ，胡 克 ，章非敏 1，2，3*
4
4
南京医科大学附属口腔医院口腔修复科，江苏省口腔疾病研究重点实验室，江苏省口腔转化医学工程研究中心，江苏
1 2 3
南京 210029；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
4

［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，0~100 μg/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。
［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
［中图分类号］ R318.08 ［文献标志码］ A ［文章编号］ 1007⁃4368（2023）05⁃669⁃09
doi：10.7655/NYDXBNS20230512

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

oxidative microenvironment
1，2，3 1，2，3 4 4 1，2，3*
CHEN Lu ，TANG Shijia ，ZHONG Caiying ，HU Ke ，ZHANG Feimin
1 Department of Prosthodontics，the Affiliated Stomatological Hospital of Nanjing Medical University，Jiangsu
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Province Key Laboratory of Oral Diseases，Jiangsu Province Engineering Research Center of Stomatological
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Translational Medicine，Nanjing 210029；Key Laboratory of Clinical Engineering，School of Biomedical Engineering
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and Informatics，Nanjing Medical University，Nanjing 211166，China

［Abstract］ 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⁃100 μg/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

［基金项目］ 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）
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通信作者（Corresponding author），E⁃mail：fmzhang@njmu.edu.cn