中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (10): 1522-1529.doi: 10.12307/2022.198

• 组织工程骨材料Tissue-engineered bone • 上一篇    下一篇

聚多巴胺辅助红景天苷改善微弧氧化纯钛的成骨性能

梁鹏晨1，2，3，史俊峰2，4，孙苗苗1，2，梁冬雨2，3，沙  爽2，5，易清清2，3，常  庆1，2   

  1. 1上海中医药大学研究生院，上海市   200120；2上海交通大学医学院附属第九人民医院·上海口腔医学先进技术与材料工程技术研究中心，上海市   200011
  • 收稿日期:2020-10-04 修回日期:2020-10-12 接受日期:2020-11-19 出版日期:2022-04-08 发布日期:2021-10-25
  • 通讯作者: 常庆，博士，副研究员，上海中医药大学研究生院，上海市 200120；上海交通大学医学院附属第九人民医院·上海口腔医学先进技术与材料工程技术研究中心，上海市 200011 易清清，硕士，主管技师，上海交通大学医学院附属第九人民医院·上海口腔医学先进技术与材料工程技术研究中心，上海市 200011；上海健康医学院附属嘉定区中心医院临床科研中心，上海市 201800
  • 作者简介:梁鹏晨，男，1997年生，四川省资阳市人，汉族，上海中医药大学在读硕士，主要从事中药与医学生物材料研究
  • 基金资助:
    国家自然科学基金资助项目(81670968)，项目负责人：常庆；上海市嘉定区卫健委重点项目(2020-ZD-03)，项目负责人：常庆；上海市嘉定区自然科学基金(JDKW-2020-0013)，项目负责人：易清清

Polydopamine assisted salidroside to improve bone formation on micro-arc oxidation of pure titanium

Liang Pengchen1, 2, 3, Shi Junfeng2, 4, Sun Miaomiao1, 2, Liang Dongyu2, 3, Sha Shuang2, 5, Yi Qingqing2, 3, Chang Qing1, 2   

  1. 1Graduate School, Shanghai University of Traditional Chinese Medicine, Shanghai 200120, China; 2Shanghai Engineering Research Center of Advanced Dental Technology and Materials, Shanghai Ninth People’s Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
  • Received:2020-10-04 Revised:2020-10-12 Accepted:2020-11-19 Online:2022-04-08 Published:2021-10-25
  • Contact: Chang Qing, MD, Associate researcher, Graduate School, Shanghai University of Traditional Chinese Medicine, Shanghai 200120, China; Shanghai Engineering Research Center of Advanced Dental Technology and Materials, Shanghai Ninth People’s Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
  • About author:Liang Pengchen, Master candidate, Graduate School, Shanghai University of Traditional Chinese Medicine, Shanghai 200120, China; Shanghai Engineering Research Center of Advanced Dental Technology and Materials, Shanghai Ninth People’s Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China; Clinical Research Center, Jiading District Central Hospital Affiliated to Shanghai University of Medicine & Health Sciences, Shanghai 201800, China
  • Supported by:
    he National Natural Science Foundation of China, No. 81670968 (to CQ); Key Project of Shanghai Jiading District Health Commission, No. 2020-ZD-03 (to CQ); the Natural Science Foundation of Jiading District of Shanghai, No. JDKW-2020-0013 (to YQQ)

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摘要： 文题释义：
红景天苷：是从红景天中提取的主要活性化合物，具有多种药理作用，近年来研究发现红景天苷对细胞成骨分化具有较好的促进作用。
聚多巴胺：一种被广泛用于生物材料表面生物功能改性中的黏合剂。多巴胺在碱性环境下可自聚合成聚多巴胺，结合于材料表面的聚多巴胺具有大量吸附包括生长因子在内的活性蛋白和小分子化合物的能力。

背景：微弧氧化技术目前已被较多用于提高钛植入体骨整合性的研究中，在钛表面构建出多种含不同活性元素的多孔涂层。近年来研究发现，红景天苷具有较好的促成骨活性。聚多巴胺作为材料表面优良的黏合剂和二次吸附的载体具有较好负载活性分子的特性。
目的：为进一步提高钛表面微弧氧化涂层的促成骨能力，在多孔形貌表面构建骨整合性优良的复合涂层。
方法：在纯钛表面制作微弧氧化涂层，记为MAO组；进一步在微弧氧化涂层表面制作聚多巴胺涂层，记为PDA组；在聚多巴胺涂层表面加载1，2，4 g/L 的红景天苷，依次记为Sal-1组、Sal-2组、Sal-4组，利用扫描电镜及X射线能谱仪分析各组涂层的微观形貌、表面元素含量及分布；通过CCK-8及荧光染色实验评估各组涂层表面的细胞增殖活性，通过碱性磷酸酶染色、茜素红染色实验观察各组涂层的促成骨分化及矿化能力；利用计算机分子对接技术预测红景天苷分子的潜在促成骨靶点。
结果与结论：①扫描电镜显示，5组试件表面为多孔微米形貌结构，平均孔径相似；X射线能谱仪分析显示，通过聚多巴胺载体可将红景天苷加载到微弧氧化涂层表面；②CCK-8实验显示，Sal-2组、Sal-4组培养第3，5天的MC3T3-E1细胞存活率高于MAO组(P < 0.05)；荧光染色实验显示，5组试件表面MC3T3-E1间形态差异较明显，与MAO组相比，Sal-2组、Sal-4组细胞形态更好、铺展面积大，与相邻细胞连接较好；③Sal-2组、Sal-4组培养第4，7天的的碱性磷酸酶活性高于MAO组(P < 0.05)；培养21 d时的茜素红染色显示，各组均有一定程度的钙结节出现，其中Sal-2组、Sal-4组的钙结节数量更加明显；④计算机分子对接结果表明，细胞外调节蛋白激酶2、c-Jun氨基末端激酶3、雌激素受体β、孕激素受体可能是红景天苷发挥促成骨作用时活性较强的靶点；⑤结果表明，通过聚多巴胺将红景天苷负载到钛微弧氧化涂层表面，适当加载量的红景天苷较好地改善了微弧氧化涂层的促成骨活性。

关键词: 材料, 纯钛, 表面改性, 微弧氧化, 聚多巴胺, 红景天苷, 成骨活性, 分子对接

Abstract: BACKGROUND: Micro-arc oxidation technology has been used to improve the osseointegration of titanium implants. A variety of porous coatings containing different active elements are constructed on the surface of titanium. In recent years, studies have found that salidroside has good bone-promoting activity. Polydopamine, as an excellent binder on the surface of the material and a carrier for secondary adsorption, has the characteristics of loading active molecules.  
OBJECTIVE: To further improve the bone-promoting ability of the micro-arc oxidation coating on the titanium surface, a composite coating with excellent osseointegration was constructed on the porous surface.
METHODS:  The micro-arc oxidation coating was made on the surface of pure titanium, which was marked as MAO group, and the polydopamine coating was further prepared on the surface of micro-arc oxidation coating, which was marked as PDA group. Salidroside (1, 2, 4 g/L) was loaded on the surface of polydopamine coating, which was divided into Sal-1 group, Sal-2 group and Sal-4 group. The micro-morphology, surface element content and distribution of each coating were analyzed by scanning electron microscope and X-ray energy dispersive spectrometer. The proliferation activity of cells on the surface of each group was evaluated by CCK-8 assay and fluorescence staining, and the abilities of bone differentiation and mineralization were observed by alkaline phosphatase staining and alizarin red staining. The potential bone-promoting targets of salidroside were predicted by computer molecular docking technique.  
RESULTS AND CONCLUSION: (1) Scanning electron microscope showed that the surface of the five groups of specimens had porous micron morphology and the average pore diameter was similar, and X-ray energy dispersive spectrometer analysis showed that salidroside could be loaded on the surface of micro-arc oxidation coating by polydopamine carrier. (2) CCK-8 experiment showed that the survival rate of MC3T3-E1 cells in Sal-2 group and Sal-4 group was higher than that in MAO group on the 3rd and 5th days of culture (P < 0.05). Fluorescence staining showed that there were significant differences in MC3T3-E1 morphology among the five groups, and compared with MAO group, Sal-2 group and Sal-4 group had better cell morphology, larger cell spreading area and better connection with neighboring cells. (3) The activity of alkaline phosphatase in Sal-2 group and Sal-4 group was higher than that in MAO group on the 4th and 7th days (P < 0.05). Alizarin red staining showed that calcium nodules appeared in all groups on the 21st day, especially in Sal-2 group and Sal-4 group. (4) Computer molecular docking results show that extracellular regulated protein kinases 2, c-Jun N-terminal kinase 3, estrogen receptor β, and progesterone receptor may be the more active targets for salidroside to promote bone formation. (5) The results showed that salidroside was loaded on the surface of titanium micro-arc oxidation coating by polydopamine, and the bone-promoting activity of micro-arc oxidation coating was improved by proper amount of salidroside.

Key words: material, pure titanium, surface modification, micro-arc oxidation, polydopamine, salidroside, osteogenic activity, molecular docking

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