Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, Brazil.
Caries Res. 2018;52(4):288-296. doi: 10.1159/000485390. Epub 2018 Jan 27.
This study detected changes in the protein profile of the acquired enamel pellicle (AEP) formed in vivo after rinsing with whole milk, fat-free milk, or water. Nine subjects in good oral condition took part in the study. The acquired pellicle was formed in the morning, for 120 min, after prophylaxis with pumice. Following this, the volunteers rinsed with 10 mL of whole milk, fat-free milk, or deionized water for 30 s, following a blinded crossover protocol. After 60 min, the pellicle was collected with filter paper soaked in 3% citric acid and processed for analysis by liquid chromatography-electrospray ionization tandem mass spectrometry. The obtained tandem mass spectrometry spectra were searched against a human protein database (Swiss-Prot). The proteomic data related to protein quantification were analysed using the PLGS software. A total of 260 proteins were successfully identified in the AEP samples collected from all groups. Forty-nine were common to all 3 groups, while 72, 62, and 49 were specific to the groups rinsing with whole milk, fat-free milk, and water, respectively. Some were typical components of the AEP, such as cystatin-B, cystatin-SN, isoforms of α-amylase, IgA and IgG, lysozyme C, protein S100 A78, histatin-1, proline-rich protein 27, statherin, and lactotransferrin. Other proteins are not commonly described as part of the AEP but could act in defence of the organism against pathogens. Distinct proteomic profiles were found in the AEP after rinsing with whole or fat-free milk, which could have an impact on bacterial adhesion and tooth dissolution. The use of fat-free milk could favourably modulate the adhesion of bacteria to the AEP as well as biofilm formation when compared with whole milk.
本研究检测了用全脂奶、脱脂奶或水漱口后体内获得性 enamel pellicle (AEP) 的蛋白质谱变化。9 名口腔状况良好的受试者参加了这项研究。在用浮石进行预防后，于上午形成获得性牙斑，持续 120 分钟。之后，志愿者们按照盲法交叉方案用 10 mL 全脂奶、脱脂奶或去离子水漱口 30 秒。60 分钟后，用滤纸收集牙斑，滤纸用 3%柠檬酸浸透，用液相色谱-电喷雾串联质谱法进行分析。获得的串联质谱图谱与人类蛋白质数据库（Swiss-Prot）进行比对。使用 PLGS 软件分析与蛋白质定量相关的蛋白质组学数据。从所有组收集的 AEP 样本中成功鉴定出 260 种蛋白质。其中 49 种在 3 组中均有，而 72、62 和 49 种分别为用全脂奶、脱脂奶和水漱口的组所特有。有些是 AEP 的典型成分，如胱抑素-B、胱抑素-SN、α-淀粉酶同工型、IgA 和 IgG、溶菌酶 C、S100A78 蛋白、组蛋白-1、富含脯氨酸的蛋白 27、牙本质蛋白、乳转铁蛋白。其他蛋白质通常不被描述为 AEP 的一部分，但可能在机体防御病原体方面发挥作用。用全脂奶或脱脂奶漱口后，AEP 的蛋白质谱明显不同，这可能会影响细菌黏附和牙齿溶解。与全脂奶相比，脱脂奶的使用可以更好地调节细菌对 AEP 的黏附以及生物膜的形成。
