Carbodiimide Inactivation of MMPs and Effect on Dentin Bonding
Carbodiimide Inactivation of MMPs and Effect on Dentin Bonding
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DOI：
10.1177/0022034513516465
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发表时间：
2014-03-01
影响因子：
7.6
通讯作者：
Breschi, L.
中科院分区：
文献类型：
--
作者：
Mazzoni, A.;Apolonio, F. M.;Breschi, L.
最近提出在粘合过程中使用蛋白质交联剂来改善粘合耐久性。本研究旨在利用酶谱和原位酶谱技术评价1-乙基-3-（3-二甲氨基丙基）碳二亚胺（EDC）交联剂抑制基质金属蛋白酶（MMP）活性的能力。测试的假设是：（1）粘合程序增加牙本质明胶分解活性和（2）EDC预处理防止这种酶活性。酶谱分析是对从经Optibond FL或Scotchbond 1XT处理的牙本质粉末中获得的蛋白质提取物进行的，所述牙本质粉末具有或不具有0.3M EDC预处理。对于原位酶谱法，使用相同的粘合剂创建粘合剂/牙本质界面，所述粘合剂应用于酸蚀刻牙本质板，所述牙本质板用EDC调节剂预处理或未处理。酶谱分析显示，应用粘合剂后牙本质内源性MMP-2和MMP-9的表达增加，而使用EDC作为引物使牙本质明胶酶失活。原位酶谱分析结果表明，所测试的粘合剂的混合层表现出强烈的胶原溶解活性，而几乎没有检测到荧光信号时，标本用EDC预处理。本研究中使用的相关性分析表明，EDC可以促进内源性牙本质MMPs在由蚀刻-冲洗粘合剂形成的混合层中的粘附。
The use of protein cross-linking agents during bonding procedures has been recently proposed to improve bond durability. This study aimed to use zymography and in situ zymography techniques to evaluate the ability of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) cross-linker to inhibit matrix metalloproteinase (MMP) activity. The hypotheses tested were that: (1) bonding procedures increase dentin gelatinolytic activity and (2) EDC pre-treatment prevents this enzymatic activity. The zymographic assay was performed on protein extracts obtained from dentin powder treated with Optibond FL or Scotchbond 1XT with or without 0.3M EDC pre-treatment. For in situ zymography, adhesive/dentin interfaces were created with the same adhesives applied to acid-etched dentin slabs pre-treated or not with EDC conditioner. Zymograms revealed increased expression of dentin endogenous MMP-2 and -9 after adhesive application, while the use of EDC as a primer inactivated dentin gelatinases. Results of in situ zymograpy showed that hybrid layers of tested adhesives exhibited intense collagenolytic activity, while almost no fluorescence signal was detected when specimens were pre-treated with EDC. The correlative analysis used in this study demonstrated that EDC could contribute to inactivate endogenous dentin MMPs within the hybrid layer created by etch-and-rinse adhesives.