中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (4): 516-520.doi: 10.12307/2022.085

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

上调骨形态发生蛋白2促进成骨细胞增殖分化的硅酸钙类材料Biodentine

杨斯迪1，2，王  倩1，许  诺3，王榕焓1，靳传奇1，陆  颖1，董  明1   

  1. 1大连医科大学口腔医学院，辽宁省大连市   116044； 2沈阳和平思麦尔口腔门诊部，辽宁省沈阳市   110000；3大连医科大学中山学院，辽宁省大连市   116000
  • 收稿日期:2021-01-06 修回日期:2021-01-08 接受日期:2021-02-05 出版日期:2022-02-08 发布日期:2021-11-03
  • 通讯作者: 董明，博士，大连医科大学口腔医学院，辽宁省大连市 116044
  • 作者简介:杨斯迪，女，1988年生，辽宁省沈阳市人，汉族，硕士，主要从事牙体牙髓病研究。 王倩，女，1985年生，辽宁省大连市人，汉族，硕士，讲师，主要从事龋病的病因与防治研究。
  • 基金资助:
    国家青年科学基金项目(81700962)，项目负责人：陆颖

Biodentine enhances the proliferation and differentiation of osteoblasts through upregulating bone morphogenetic protein-2

Yang Sidi1, 2, Wang Qian1, Xu Nuo3, Wang Ronghan1, Jin Chuanqi1, Lu Ying1, Dong Ming1   

  1. 1School of Stomatology, Dalian Medical University, Dalian 116044, Liaoning Province, China; 2Shenyang Heping Simaier Dental Clinic, Shenyang 110000, Liaoning Province, China; 3Zhongshan College of Dalian Medical University, Dalian 116000, Liaoning Province, China
  • Received:2021-01-06 Revised:2021-01-08 Accepted:2021-02-05 Online:2022-02-08 Published:2021-11-03
  • Contact: Dong Ming, PhD, School of Stomatology, Dalian Medical University, Dalian 116044, Liaoning Province, China
  • About author:Yang Sidi, Master, School of Stomatology, Dalian Medical University, Dalian 116044, Liaoning Province, China; Shenyang Heping Simaier Dental Clinic, Shenyang 110000, Liaoning Province, China Wang Qian, Master, Lecturer, School of Stomatology, Dalian Medical University, Dalian 116044, Liaoning Province, China
  • Supported by:
    the National Youth Science Foundation Project, No. 81700962 (to LY)

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摘要：

文题释义：
Biodentine：属于硅酸钙类材料，主要由硅酸三钙、碳酸钙和氧化锆组成，具有硅酸钙的生物和物理特性，被认为是效果较佳的根管穿孔修复材料。与传统的硅酸钙类材料三氧化矿化物凝聚体相比，Biodentine在理化性质、生物学功能及操作时间等方面效果更佳。Biodentine具有类似于牙本质的机械性能，可以用于牙冠及牙根部位的牙本质修复，作为牙髓腔垫底材料也可用于直接盖髓，以及作为封闭牙髓与牙周膜的修补材料。
骨形态发生蛋白2：属于转化生长因子β超家族的成员，是有效的骨诱导细胞因子，在骨修复中起关键作用。
背景：Biodentine是一种具有生物相容性的新型硅酸钙口腔修复材料，它不仅可以增加牙髓细胞中生长因子的分泌，也可以诱导牙本质的矿化，但是其在骨修复方面的研究较少。
目的：通过构建干扰骨形态发生蛋白2的小鼠成骨前体细胞株，研究Biodentine对于干扰骨形态发生蛋白2小鼠成骨前体细胞株的增殖及分化能力的影响。
方法：将0.1，1，10，100 g/L 的Biodentine浸提液作用于小鼠成骨前体细胞24 h，通过CCK-8与碱性磷酸酶活性实验筛选最佳的作用质量浓度10 g/L，用于以下实验。将Biodentine浸提液作用于小鼠成骨前体细胞1，3，5，7 d，以未干预组为对照，通过CCK-8与碱性磷酸酶活性实验观察作用前后细胞的增殖及成骨分化。构建干扰骨形态发生蛋白2的小鼠成骨前体细胞株，通过CCK-8与碱性磷酸酶活性实验观察干扰前后细胞的增殖及成骨分化。构建干扰骨形态发生蛋白2的小鼠成骨前体细胞株，Biodentine组加入10 g/L 的Biodentine浸提液，设置单纯干扰组为对照，作用24，48 h后，通过CCK-8与碱性磷酸酶活性实验观察细胞的增殖及成骨分化；作用24 h后，Real-time PCR、Western blot检测碱性磷酸酶mRNA与蛋白的表达。
结果与结论：①10 g/L的Biodentine浸提液在1，3，5，7 d可促进小鼠成骨前体细胞的增殖与成骨分化；②干扰骨形态发生蛋白2可抑制小鼠成骨前体细胞的增殖与成骨分化；③Biodentine组作用24，48 h后的细胞增殖与成骨分化能力高于干扰组(P < 0.05)，作用24 h后的碱性磷酸酶mRNA与蛋白表达高于干扰组(P < 0.05)；④结果表明，10 g/L的Biodentine可以通过骨形态发生蛋白2增强成骨细胞的增殖及成骨分化。

https://orcid.org/0000-0002-8762-7644 (杨斯迪) 

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

关键词: Biodentine, 硅酸钙材料, 成骨前体细胞, MC3T3-E1细胞, 骨形态发生蛋白2, 细胞增殖, 细胞分化

Abstract: BACKGROUND: Biodentine is a new kind of biocompatible calcium silicate dental restoration material. It can not only increase the secretion of growth factors in dental pulp cells, but also induce the mineralization of dentin. However, the research of Biodentine in bone repair is few.
OBJECTIVE: To investigate the effects of Biodentine on the proliferation and differentiation of bone morphogenetic protein-2 interfered mouse osteoblast precursor cell lines by constructing mouse osteoblast precursor cell lines in which bone morphogenetic protein-2 was interfered. 
METHODS: The 0.1, 1, 10, and 100 g/L Biodentine extract was applied to mouse osteoblast precursor cells for 24 hours. The optimal concentration of 10 g/L was selected by CCK-8 and alkaline phosphatase for the following experiments. Biodentine extract was used to treat mouse osteoblast precursor cells for 1, 3, 5 and 7 days, and the non-intervention group was used as the control group. The cell proliferation and osteogenic differentiation were observed by CCK-8 assay and alkaline phosphatase activity test. Mouse osteoblast precursor cell lines were constructed by interfering with bone morphogenetic protein-2. The proliferation and osteogenic differentiation of cells were observed by CCK-8 assay and alkaline phosphatase activity test. The mouse osteoblast precursor cell lines interfering with bone morphogenetic protein-2 were constructed. The Biodentine group was added with 10 g/L Biodentine extract, and the non-intervention group was set as the control. After 24 and 48 hours of treatment, the proliferation and osteogenic differentiation of the cells were observed by CCK-8 assay and alkaline phosphatase activity test. After 24 hours of treatment, the expression levels of alkaline phosphatase mRNA and protein were detected by real-time PCR and western blot assay. 
RESULTS AND CONCLUSION: (1) 10 g/L Biodentine extract could promote the proliferation and osteogenic differentiation of mouse osteoblast progenitor cells at 1, 3, 5 and 7 days. (2) Interfering with bone morphogenetic protein-2 could inhibit the proliferation and osteogenic differentiation of mouse osteoblast progenitor cells. (3) The cell proliferation and osteogenic differentiation abilities of Biodentine group were higher than those of control group at 24 and 48 hours (P < 0.05), and the expression levels of alkaline phosphatase mRNA and protein of Biodentine group were higher than those of the control group (P < 0.05). (4) The results showed that 10 g/L Biodentine could enhance the proliferation and osteogenic differentiation of osteoblasts through bone morphogenetic protein-2.

Key words: Biothentine, calcium silicate material, osteoblast precursor cell, MC3T3-E1 cell, bone morphogenetic protein-2, cell proliferation, cell differentiation

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