中国组织工程研究

中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (18): 2843-2848.doi: 10.3969/j.issn.2095-4344.0776

• 组织工程口腔材料 tissue-engineered oral materials • 上一篇    下一篇

生物活性牙本质替代物Biodentine对MG-63细胞增殖及成骨作用的影响

翟姣妍1，王丽娜2，刘启成2，史 春2，牛卫东2   

  1. 1大连医科大学口腔医学院附属口腔医院，辽宁省大连市 116023；2大连医科大学口腔医学院，辽宁省大连市 116044
  • 收稿日期:2018-01-28 出版日期:2018-06-28 发布日期:2018-06-28
  • 通讯作者: 牛卫东，博士，教授，大连医科大学口腔医学院，辽宁省大连市 116044
  • 作者简介:翟姣妍，女，1987年生，河南省开封市人，汉族，2015年大连医科大学毕业，硕士，主要从事牙体牙髓病学的临床与基础研究。
  • 基金资助:

    国家自然科学基金(81171538)

Effects of Biodentine on proliferation and osteogenesis of MG-63 cells

Zhai Jiao-yan1, Wang Li-na2, Liu Qi-cheng2, Shi Chun2, Niu Wei-dong2   

  1. 1Hospital of Stomatology, School of Stomatology, Dalian Medical University, Dalian 116023, Liaoning Province, China; 2College of Stomatology, Dalian Medical University, Dalian 116044, Liaoning Province, China
  • Received:2018-01-28 Online:2018-06-28 Published:2018-06-28
  • Contact: Niu Wei-dong, M.D., Professor, College of Stomatology, Dalian Medical University, Dalian 116044, Liaoning Province, China
  • About author:Zhai Jiao-yan, Master, Hospital of Stomatology, School of Stomatology, Dalian Medical University, Dalian 116023, Liaoning Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81171538

PDF

173

摘要：

文章快速阅读：

 

文题释义：
生物活性物质Biodentine：是2009年由法国赛普敦公司研发的一种生物活性牙本质替代物，由粉、液混合聚合后使用，粉剂中主要成分为硅酸三钙，并含有硅酸二钙、碳酸钙、氧化锆、氧化铁，其中硅酸三钙是其主要成分；液剂由氯化钙和水溶性聚合物组成；其改善了矿物三氧化凝聚体的一些如操作不便、聚合时间长、机械性能低的潜在缺点，成为国内外学者们关注的主要产品之一。
细胞的成骨活性：矿化结节的形成能力作为反映成骨细胞晚期分化的指标，代表着成骨细胞分化成熟、进入到行使功能的阶段。钙离子水平变化是骨细胞增殖分化、骨组织成骨潜能的标志之一。茜素红和钙离子以鳌和方式形成复合物，产生红色矿化结节，矿化结节数目可反映成骨细胞的矿化程度。
 
背景：生物活性物质Biodentine具有良好的抗压强度、粘接强度及较少的微渗漏等诸多优点，已被成功应用于多种临床适应证，然而有关Biodentine是否可促进成骨细胞成骨的研究罕见。
目的：研究不同浓度Biodentine对人骨肉瘤细胞MG-63增殖及成骨作用的影响。
方法：制备不同浓度梯度(1、1/2、1/4、1/8和1/16)的Biodentine浸提液。将人骨肉瘤细胞MG-63分6组培养，分别加入MEM 培养液(对照组)及5种浓度的Biodentine浸提液，采用CCK-8法检测培养第1，3，5，7天的细胞增殖，筛选Biodentine浸提液最佳浓度。将人骨肉瘤细胞MG-63分2组培养，分别加入MEM 培养液(空白对照组)及最佳浓度的Biodentine材料浸提液(实验组)，培养第1，3，5，7天，采用Real-time PCR法检测细胞中成骨因子Runx2 mRNA表达；培养第10，14天行茜素红染色，观察细胞钙化结节形成情况。

结果与结论：①培养第1，3天时，不同浓度Biodentine材料浸提液组活细胞数量与对照组相比无差异；培养第5天，1浓度材料浸提液组活细胞数量明显低于对照组(P < 0.05)，1/2、1/4、1/8浓度材料浸提液组活细胞数量与对照组比较无差异，1/16浓度材料浸提液组活细胞数量明显高于对照组(P < 0.05)，因此实验选择1/16浓度材料浸提液进行后续实验；②实验组培养第1，3，5，7天的Runx2 mRNA表达量分别为空白对照组的1.14，5.29，1.08，2.11倍(P均< 0.05)；③培养第10天时，实验组与空白对照组均未见矿化结节；培养第14天，两组均可见矿化结节，实验组的面积更大、颜色更深；④结果表明在一定浓度下，Biodentine可促进人骨肉瘤细胞MG-63增殖及成骨活性。

ORCID: 0000-0001-8915-7335(翟姣妍) 

关键词: Biodentine, MG-63细胞, 细胞增殖, 成骨作用, 国家自然科学基金, 生物材料

Abstract:

BACKGROUND: As a bioactive substance, Biodentine has good compressive strength, bonding strength and less micro-leakage. It has been successfully applied to a variety of clinical indications. However, much less is known about whether Biodentine can promote osteogenesis.

OBJECTIVE: To explore the effects of Biodentine of different concentrations on proliferation and osteogenesis of MG-63 cells.
METHODS: The extracts of Biodentine with different concentration gradients (1, 1/2, 1/4, 1/8, 1/16) were prepared. MG-63 cells were cultured in six groups, with the addition of minimum essential medium (control group) and five concentrations of Biodentine extracts. Cell proliferation was detected by cell counting kit-8 assay at 1, 3, 5 and 7 days, and then the best concentration of Biodentine extract was screened. Another MG-63 cells were cultured in minimum essential medium (blank control group) and Biodentine extract of the optimal concentration (experimental group). The expression of osteogenic factor Runx2 mRNA in MG-63 cells was detected by real-time PCR at 1, 3, 5 and 7 days of culture. Then alizarin red staining was used to observe calcified nodules in MG-63 cells at 10 and 14 days of culture.
RESULTS AND CONCLUSION: (1) At 1 and 3 days of culture, the number of viable cells in different concentration groups was similar to that in the control group (P > 0.05). At 5 days of culture, compared with the control group, the number of viable cells in MG-63 cells was significantly lowered in the 1 concentration group, showed no significant changes in the 1/2, 1/4, 1/8 concentration groups (P > 0.05), and was significantly increased in the 1/16 concentration group (P < 0.05). Therefore, the 1/16 concentration of Biodentine extract was used for further experiment. (2) Runx2 mRNA expression of the experimental group at 1, 3, 5 and 7 days of culture was 1.14, 5.29, 1.08 and 2.11 times that of the control group, respectively (all P < 0.05). (3) At 10 days of culture, both the experimental group and blank control group showed no mineralized nodules; at 14 days of culture, mineralized nodules were observed in the two groups, and larger and darker nodules were seen in the experiment group. To conclude, Biodentine at certain concentrations can promote the proliferation and osteogenic activity of human osteosarcoma cell line MG-63. 

中图分类号: