Caries Res. 2018;52(5):367-377. doi: 10.1159/000486969. Epub 2018 Mar 6.
Changes in the protein profile of acquired enamel pellicles (AEP) formed in vivo over different time periods were evaluated after the application of hydrochloric acid (HCl).
Nine subjects were submitted to dental prophylaxis with pumice. After 3 or 120 min, the teeth were isolated with cotton rolls and 50 μL of 0.1 M HCl (pH 1.0), 0.01 M HCl (pH 2.0), or deionized water were applied on the buccal surface of the teeth for 10 s. The AEP was then collected using an electrode filter paper presoaked in 3% citric acid. After protein extraction, the samples were submitted to reverse-phase liquid chromatography coupled to mass spectrometry (nano LC-ESI-MS/MS). Label-free quantification was performed (Protein Lynx Global Service software).
A total of 180 proteins were successfully identified in the AEP samples. The number of identified proteins increased with the time of pellicle formation. Only 4 proteins were present in all the groups (isoforms of IgA, serum albumin, and statherin). The greatest number of proteins identified uniquely in one of the groups was obtained for the groups treated with HCl after 2 h of pellicle formation (approx. 50 proteins).
Proteins resistant to removal by HCl, such as serum albumin and statherin, were identified even in the short-term AEP. In addition, 120-min pellicles present many proteins that are resistant to removal by HCl. This suggests an increase in protection against intrinsic acids with the time of pellicle formation, which should be evaluated in future studies.
评估盐酸(HCl)作用后不同时间形成的获得性牙釉质获得性牙釉质(AEP)蛋白图谱的变化。
9 名受试者接受浮石牙洁治。3 或 120 分钟后,用棉卷隔离牙齿,在牙齿的颊面用 50μL 0.1M HCl(pH 1.0)、0.01M HCl(pH 2.0)或去离子水作用 10s。然后使用预先用 3%柠檬酸浸泡的电极滤纸收集 AEP。在进行蛋白质提取后,将样品提交进行反相液相色谱-电喷雾质谱联用(nano LC-ESI-MS/MS)。采用无标记定量法(Protein Lynx Global Service 软件)进行分析。
在 AEP 样本中成功鉴定出 180 种蛋白质。形成牙斑时间越长,鉴定出的蛋白质数量就越多。所有组中都存在 4 种蛋白质(IgA 同工型、血清白蛋白和唾液素)。在形成牙斑 2 小时后用 HCl 处理的组中,唯一鉴定出的蛋白质数量最多(约 50 种)。
即使是在短期的 AEP 中,也能识别出对 HCl 去除具有抗性的蛋白质,如血清白蛋白和唾液素。此外,120 分钟的牙斑具有许多对 HCl 去除具有抗性的蛋白质。这表明随着牙斑形成时间的增加,对内在酸的保护作用增加,这应该在未来的研究中进行评估。