机构地区:[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
出 处:《中国组织工程研究》2018年第18期2843-2848,共6页Chinese Journal of Tissue Engineering Research
基 金:国家自然科学基金(81171538)~~
摘 要: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, bot
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
分 类 号:R318[医药卫生—生物医学工程]
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