不同粘结条件下健康成人牙本质中MMP-2和MMP-9表达水平及其意义

doi:10.13481/j.1671-587x.20180612

摘要/Abstract

摘要： 目的：探讨不同粘结条件下健康成人牙本质中基质金属蛋白酶2（MMP-2）和基质金属蛋白酶9（MMP-9）表达水平，为临床应用基质金属蛋白酶（MMPs）抑制剂提高粘结效果提供依据。方法：收集20颗成人新鲜离体磨牙，液氮冷却条件下将其研磨为牙本质粉；模拟口内条件，对牙本质分别进行自酸蚀粘结（Single Bond Universal）和全酸蚀粘结（35%磷酸凝胶+Adpter Single Bond 2）处理，以不进行任何处理的牙本质作为阴性对照组，10%磷酸溶液（自酸蚀粘结）或10%磷酸溶液+35%磷酸凝胶处理的脱矿牙本质粉（全酸蚀粘结）处理的牙本质作为阳性对照组，只进行Single Bond Universal （自酸蚀粘结）处理和35%磷酸凝胶+Adpter Single Bone2（全酸蚀粘结）处理的牙本质作为空白对照组；在粘结过程中分别使用MMPs抑制剂氯己定（CHX）和米诺环素（MI）预处理，作为CHX预处理组和MI预处理组；采用10%次氯酸钠进行牙本质老化，作为老化组，并在老化组基础上加入CHX和MI作为CHX预处理老化组和MI预处理老化组。采用明胶酶谱法进行聚丙烯酰胺凝胶电泳，孵育染色脱色后，采用凝胶图像分析系统分析条带，计算各组牙本质中MMP-2和MMP-9表达水平。结果：在自酸蚀粘结条件下，与空白对照组比较，CHX预处理组牙本质中MMP-2和MMP-9表达水平降低（P<0.05），MI预处理组牙本质中MMP-2和MMP-9表达水平降低（P<0.05）；与空白老化对照组比较，CHX预处理老化组牙本质中MMP-2和MMP-9表达水平降低（P<0.05），MI预处理老化组牙本质中MMP-2和MMP-9表达水平降低（P<0.05）。在全酸蚀粘结条件下，与空白对照组比较，CHX预处理组和MI预处理组牙本质中MMP-2表达水平降低（P<0.05）；与空白老化对照组比较，CHX预处理老化组与MI预处理老化组牙本质中MMP-2表达水平降低（P<0.05）。结论：在牙本质粘结过程中，CHX和MI能够通过降低MMP-2和MMP-9表达水平减缓酶解反应，从而增强粘结强度。

关键词: 牙本质, 粘结, 基质金属蛋白酶, 基质金属蛋白酶抑制剂

Abstract: Objective: To investigate the expression levels of matrix metalloproteinase 2(MMP-2) and matrix metalloproteinase 9(MMP-9) in dentin of healthy adults under different adhesion conditions, and to provide the basis for improving the binding effect of matrix metalloproteinases(MMPs) inhibitors in clinic.Methods: A total of 20 adult freshlyextracted molars were collected and ground into dentin powder under liquid nitrogen cooling.The oral condition was simulated.The dentin was treated with self-etching bonding (Single Bond Universal) and total-etching bonding (35% phosphate gel +Adpter Single Bond 2). The dentin without any treatment was regarded as negative control group, the dentin treated with 10% phosphoric acid(self-etching bonding)or 10% phosphoric acid+ 35% phosphate gel (total-etching bonding) was regarded as positive control group, the dentin treated with Single Bond Universal(self-etching bonding) or 35% phosphate gel+Adper Single Bond 2 (total-etching bonding) was regarded as blank control group; the dentin pretreated with the MMPs inhibitors chlorhexidine(CHX) and minocycline(MI) during the bonding process respectively was regarded as CHX group and MI group,and the dentin treated with 10% sodium hypochlorite was regarded as aging group; on the basis of aging group, the dentin treated with CHX and MI was regarded as CHX aging group and MI aging group.Gelatin zymography was used to perform the polyacrylamide gel electrophoresis; after incubating, staining and decoloring, the bands were analyzed by gel image analysis system, and the expression levels of MMP-2 and MMP-9 in dentin were calculated.Results: Under the condition of self-etching bonding, compared with blank control group, the expression levels of MMP-2 and MMP-9 in dentin in CHX group were decreased (P<0.05), and the expression levels of MMP-2 and MMP-9 in the dentin powder in MI group were decreased (P<0.05);compared with aging blank control group, the expression levels of MMP-2 and MMP-9 in dentin in CHX aging group were decreased (P<0.05), and the expression levels of MMP-2 and MMP-9 in dentin in MI aging group were decreased (P<0.05). Under the condition of total-etching bonding, compared with blank control group, the expression levels of MMP-2 in dentin in CHX group and MI group were decreased (P<0.05); compared with aging blank control group, the expression levels of MMP-2 in dentin in CHX aging group and MI aging group were decreased (P<0.05).Conclusion: In the process of dentin bonding, CHX and MI could slow down the enzymatic reaction and improve the bonding strength by decreasing the expression levels of MMP-2 and MMP-9.

Key words: dentin, bonding, matrix metalloproteinase, matrix metalloproteinase inhibitor

中图分类号:

R783.1

王春萌, 李贺, 张志鹰, 李男男, 赵聪, 洪丽华, 张志民. 不同粘结条件下健康成人牙本质中MMP-2和MMP-9表达水平及其意义[J]. 吉林大学学报(医学版), 2018, 44(06): 1179-1184.

WANG Chunmeng, LI He, ZHANG Zhiying, LI Nannan, ZHAO Cong, HONG Lihua, ZHANG Zhimin. Expression levels of MMP-2 and MMP-9 in dentin of healthy adults under different bonding conditions and their significances[J]. Journal of Jilin University(Medicine Edition), 2018, 44(06): 1179-1184.

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