Biodegradation of composite resin with ester linkages: Identifying human salivary enzyme activity with a potential role in the esterolytic process
Biodegradation of composite resin with ester linkages: Identifying human salivary enzyme activity with a potential role in the esterolytic process
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DOI：
10.1016/j.dental.2014.05.031
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发表时间：
2014-08-01
期刊：
影响因子：
5
通讯作者：
Santerre, J. Paul
中科院分区：
文献类型：
--
作者：
Cai, Kuihua;Delaviz, Yasaman;Santerre, J. Paul
目标。牙科树脂单体（如双酚a -甲基丙烯酸缩水甘油酯（BisGMA）和三甘醇二甲基丙烯酸酯（TEGDMA））中的酯键容易被唾液酯酶水解降解，但对这一过程中涉及的特定酯酶活性知之甚少。这项工作的目的是从唾液中分离和鉴定与酯酶活性相关的显性蛋白，这些蛋白被证明参与了bisgma的降解。采集人全唾液，在HiPrep 16/60 Sephacryl S-200 HR柱分离前进行处理。通过阴离子交换柱（Mono-Q (10/100G)）进一步分离酯酶活性最高的部分。分离的部分然后通过凝胶电泳分离，并与常见的bench marker酯酶，胆固醇酯酶（CE）和已报道表达酯酶活性的商业白蛋白进行比较。用质谱法对怀疑含有酯酶活性的蛋白进行了分析。使用市售的类似于MS鉴定的唾液酯酶蛋白的蛋白质来复制酶复合物并确认其对bisgma的降解活性。质谱分析结果表明，酯酶活性最高的酶组分为白蛋白、zn -a2糖蛋白、a-淀粉酶、TALD01蛋白、转铁蛋白、脂酰化蛋白2和催乳素诱导蛋白。研究得出结论，每个部分凝胶上的主要酯酶带与CE活性不重叠，白蛋白活性成为相对于BisGMA降解具有显著酯酶活性的主要候选物，特别是当它在微碱性条件下与zn -a2糖蛋白形成复合物时。这些酶复合物可以作为生理相关的配方来测试复合树脂的生物稳定性。(C) 2014年牙科材料学会。Elsevier Ltd.出版。版权所有。
Objectives. The ester linkages contained within dental resin monomers (such as Bisphenol A-glycidylmethacrylate (BisGMA) and triethylene glycol dimethacrylate (TEGDMA)) are susceptible to hydrolytic degradation by salivary esterases, however very little is known about the specific esterase activities implicated in this process. The objective of this work was to isolate and identify the dominant proteins from saliva that are associated with the esterase activities shown to be involved in the degradation of BisGMA.Methods. Human whole saliva was collected and processed prior to separation in a HiPrep 16/60 Sephacryl S-200 HR column. The fraction with the highest esterase activity was further separated by an anion exchange column (Mono-Q (10/100G)). Isolated fractions were then separated by gel electrophoresis, and compared to a common bench marker esterase, cholesterol esterase (CE), and commercial albumin which has been reported to express esterase activity. Proteins suspected of containing esterase activity were analyzed by Mass Spectroscopy (MS). Commercially available proteins, similar to the salivary esterase proteins identified by MS, were used to replicate the enzymatic complexes and confirm their degradation activity with respect to BisGMA.Results. MS data suggested that the enzyme fraction with the highest esterase activity was contained among a group of proteins consisting of albumin, Zn-a2-glycoprotein, a-amylase, TALD01 protein, transferrin, lipocalin2, and prolactin-induced protein. Studies concluded that the main esterase bands on the gels in each fraction did not overlap with CE activity, and that albumin activity emerged as a lead candidate with significant esterase activity relative to BisGMA degradation, particularly when it formed a complex with Zn-a2-glycoprotein, under slightly basic conditions.Signcance. These enzyme complexes can be used as a physiologically relevant formulation to test the biostability of composite resins. (C) 2014 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.