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

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

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 m RNA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 m RNA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。

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.