Proteoglycans affect monomer infiltration in the etch-and-rinse bonding technique
Proteoglycans affect monomer infiltration in the etch-and-rinse bonding technique
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蛋白聚糖影响蚀刻和冲洗键合技术中的单体渗透。
DOI：
10.1016/j.dental.2014.05.015
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发表时间：
2014-11-01
期刊：
影响因子：
5
通讯作者：
Chen, Ji-Hua
中科院分区：
文献类型：
--
作者：
Lu, Shuai;Zhao, San-Jun;Chen, Ji-Hua
目标.采用免疫荧光双标记技术检测牙本质中蛋白多糖（PG）和胶原纤维（CF）的含量，并研究两种蚀刻冲洗粘合剂对甲苯磺酰苯丙氨酸氯甲基酮（TPCK）处理的胰蛋白酶（TRY）预处理牙本质的单体浸润情况。取中冠牙本质30 μ m切片，用37%磷酸凝胶酸蚀15 s。经TRY酶切或不经TRY酶切预处理后，切片进行双重免疫荧光标记，激光共聚焦扫描显微镜（CLSM）观察。用场发射扫描电镜（FE-SEM）观察TRY处理前后脱矿牙本质基质的变化。两种蚀刻和冲洗粘合剂，Adper Single Bond 2（SB）和Prime & Bond NT（PBNT），应用于用或不用TRY预处理的牙本质表面。在共焦显微拉曼光谱下评估杂化层的厚度，并用双向方差分析进行分析。绿色和红色荧光用于表示在同一切片中共定位的具有不同分布的PG和CF。PG主要分布于牙本质小管内腔和小管周围牙本质，而I型胶原纤维主要分布于牙本质小管间和小管周围牙本质。TRY预处理组牙本质表面红色荧光减弱或消失，牙本质小管管腔内的有机丝消失，小管变大，粘接剂与TRY预处理组牙本质表面的混合层厚度显著增加（SB和PBNT均p < 0.001）。双重免疫荧光标记方法可用于研究人牙本质基质，而无需对整个牙本质碎片进行脱钙。牙本质小管腔内和小管周围牙本质中存在蛋白多糖，可抑制粘附树脂单体的浸润。TRY消化的使用增加了由所测试的两步蚀刻和冲洗粘合剂产生的混合层的厚度。(C)2014年牙科材料学院。由爱思唯尔有限公司出版。保留所有权利。
Objectives. To identify proteoglycans (PGs) and collagen fibrils (CF) within human dentin by means of a dual immunofluorescent labeling technique and to investigate the monomer infiltration of two etch-and-rinse adhesives to tosyl-phenylalanine chloromethyl-ketone(TPCK)-treated trypsin (TRY)-pretreated dentin.Methods. Thirty-micrometer sections of middle coronal dentin were obtained and etched with 37% phosphoric acid gel for 15 s. After preconditioning with or without TRY digestion, the sections were subjected to dual immunofluorescent labeling and observed witha confocal laser scanning microscope (CLSM). Demineralized dentin matrixes treated with or without TRY were observed with field emission scanning electron microscope (FE-SEM). Two etch-and-rinse adhesives, Adper Single Bond 2 (SB) and Prime & Bond NT (PBNT), were applied to the dentin surfaces that were pretreated with or without TRY. The thickness of the hybrid layers was evaluated under confocal micro-Raman spectroscopy and analyzed with a two-way ANOVA.Results. Green and red fluorescence was used to represent the PGs and the CF that were colocalized in the same section with different distributions. PGs were localized in the lumens of the dentin tubules and in peritubular dentin, while the type-I collagen fibrils were localized in intertubular dentin and peritubular dentin. After preconditioning with TRY digestion, the red fluorescence decreased or disappeared, the organic filaments in the lumens of the dentin tubules disappeared, the tubules were enlarged, and the hybrid layer thickness for adhesives bonded to the TRY-pretreated dentin surfaces were significantly increased (p < 0.001 for both SB and PBNT).Significance. The dual immunofluorescence labeling methodology can be used to study the human dentin matrix without decalcifying the entire dentin fragment. Proteoglycans were localized in the lumens of the dentin tubules and in peritubular dentin, which could depress the infiltration of the adhesive resin monomers. The use of TRY digestion increased the thickness of the hybrid layer created by the tested two-step etch-and-rinse adhesive. (C) 2014 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.