生物活性玻璃和牙本质浸提蛋白对人牙髓细胞的作用

发布时间：2018-07-16 19:49

【摘要】：目的:明确生物活性玻璃(bioactive glass,BG)和牙本质浸提蛋白(extracted dentin proteins,EDP)对人牙髓细胞增殖、成牙本质向分化、矿化的作用。方法:采用酶消化法培养牙髓细胞,传代至第4代用于实验。分别用含BG-EDP浸提液、BG浸提液、EDP浸提液的达尔伯克必需基本培养基(Dulbecco’s minimum essential medium,DMEM)培养牙髓细胞,对照组为DMEM培养组;通过细胞活性噻唑蓝比色法检测细胞增殖能力,碱性磷酸酶(alkaline phosphatase,ALP)活性检测及real-time PCR检测细胞成牙本质向分化能力,茜素红钙化结节染色及氯化十六烷基吡啶半定量分析检测细胞矿化能力。结果:BG-EDP组3、7、9 d时(光密度值1.36±0.06、2.52±0.20、2.72±0.29)能够增强人牙髓细胞增殖活性,同BG组(光密度值1.20±0.26、2.33±0.26、2.50±0.30)、EDP组(光密度值1.13±0.15、2.10±0.13、2.38±0.22)和对照组(光密度值0.84±0.17、1.84±0.18、1.95±0.19)比较差异具有统计学意义(P0.05)。7 d时,BG-EDP组ALP活性与EDP组、对照组差异无统计学意义,分化相关基因(DSPP、DMP-1)表达无明显升高。14 d时,BG-EDP组ALP活性升高(56.67±1.83),显著高于EDP组(41.98±9.71)及对照组(30.82±6.70),P0.05,但同BG组(56.29±6.20)相比差异无统计学意义(P0.05);BG-EDP组DSPP表达量明显升高(5.79±1.94),显著高于BG组(2.62±0.46)、EDP组(2.66±1.06)及对照组(1.84±0.76),P0.05;BG-EDP组DMP-1表达升高(3.87±1.87),略高于BG组(1.89±0.90)、EDP组(2.38±1.04)和对照组(2.25±0.93),但差异无统计学意义(P0.05)。诱导2周后,BG-EDP组形成较多矿化结节,氯化十六烷基吡啶半定量分析显示BG-EDP组钙化量最高(0.27±0.01)。结论:同单纯的BG及EDP相比,BG-EDP复合后具有更强的促进人牙髓细胞的增殖、成牙本质向分化、矿化作用。
[Abstract]:Aim: to investigate the effects of bioactive glassglass (BG) and dentin extraction protein (extracted dentin proteinsen on the proliferation, differentiation and mineralization of human dental pulp cells. Methods: dental pulp cells were cultured by enzyme digestion. Dental pulp cells were cultured in Dulbeccos minimum essential mediumentum (DMEM) medium containing BG-EDP extract and BG extract, respectively, and the proliferation of dental pulp cells was detected by cell activity thiazolyl blue colorimetry. The activity of alkaline phosphatase (ALP) and the ability of odontoblast differentiation were detected by real-time, alizarin red calcified nodule staining and semi-quantitative analysis of cetylpyridine chloride. Results the proliferation activity of human dental pulp cells was significantly increased in the BG-EDP group (light density 1.36 卤0.06 卤2.52 卤0.202.72 卤0.29) on the 9th day (optical density 1.20 卤0.262.33 卤0.260.50 卤0.30). The ALP activity in the BG-EDP group was significantly higher than that in the EDP group (optical density 1.13 卤0.152.10 卤0.132.38 卤0.22) and the control group (0.84 卤0.17 卤1.84 卤0.18 卤1.95 卤0.19). The ALP activity in the BG-EDP group was significantly higher than that in the EDP group on the 7th day (P 0.05). There was no significant difference between the control group and the control group. The activity of ALP in BG-EDP group (56.67 卤1.83) was significantly higher than that in EDP group (41.98 卤9.71) and control group (30.82 卤6.70) (P0.05), but there was no significant difference between BG-EDP group and BG group (56.29 卤6.20) (P0.05). The expression of DSPP in BG-EDP group was significantly higher than that in BG group (5.79 卤1.94), significantly higher than that in BG group (2.62 卤0.46). DMP-1 expression in BG-EDP group (2.66 卤1.06) and control group (1.84 卤0.76) increased (3.87 卤1.87), slightly higher than that in BG group (2.38 卤1.04) and control group (2.25 卤0.93), but there was no significant difference (P0.05). After 2 weeks of induction, more mineralized nodules were formed in BG-EDP group. The results of semi-quantitative analysis of cetylpyridine chloride showed that the highest amount of calcification was found in BG-EDP group (0.27 卤0.01). Conclusion: compared with BG and EDP alone, BG-EDP can promote the proliferation, dentin differentiation and mineralization of human dental pulp cells.
【作者单位】：北京大学口腔医学院·口腔医院牙体牙髓科口腔数字化医疗技术和材料国家工程实验室口腔数字医学北京市重点实验室;
【基金】：国家自然科学基金(51372005)资助~~
【分类号】：R783.1

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