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人牙和牛牙表面形成的获得性釉质 pellicle 的蛋白质组学分析:一项使用鲍鲁原位 pellicle 模型(BISPM)的研究

Proteomic analysis of the acquired enamel pellicle formed on human and bovine tooth: a study using the Bauru in situ pellicle model (BISPM).

作者信息

Pelá Vinícius Taioqui, Cassiano Luiza Paula Silva, Ventura Talita Mendes da Silva, Souza-E-Silva Cintia Maria de, Gironda Carlos Condarco, Rios Daniela, Buzalaf Marília Afonso Rabelo

机构信息

Universidade Federal de São Carlos, Departmento de Genetica e Evolução, São Carlos, SP, Brasil.

Faculdades Integradas Aparicio Carvalho, Porto Velho, RO, Brasil.

出版信息

J Appl Oral Sci. 2018 Dec 10;27:e20180113. doi: 10.1590/1678-7757-2018-0113.

Abstract

OBJECTIVES

The acquired enamel pellicle (AEP) is an organic film, bacteria-free, formed in vivo as a result of the selective adsorption of salivary proteins and glycoproteins to the solid surfaces exposed to the oral environment. Objective: This study aimed to compare the proteomic profile of AEP formed in situ on human and bovine enamel using a new intraoral device (Bauru in situ pellicle model - BISPM).

MATERIAL AND METHODS

Material and Methods: One hundred and eight samples of human and bovine enamel were prepared (4×4 mm). Nine subjects with good oral conditions wore a removable jaw appliance (BISPM) with 6 slabs of each substrate randomly allocated. The AEP was formed during the morning, for 120 minutes, and collected with an electrode filter paper soaked in 3% citric acid. This procedure was conducted in triplicate and the pellicle collected was processed for analysis by LC-ESI-MS/MS. The obtained mass spectrometry MS/MS spectra were searched against human protein database (SWISS-PROT).

RESULTS

Results: The use of BISPM allowed the collection of enough proteins amount for proper analysis. A total of 51 proteins were found in the AEP collected from the substrates. Among them, 15 were common to both groups, 14 were exclusive of the bovine enamel, and 22 were exclusive of the human enamel. Proteins typically found in the AEP were identified, such as Histatin-1, Ig alpha-1, Ig alpha 2, Lysozyme C, Statherin and Submaxillary gland androgen-regulated protein 3B. Proteins not previously described in the AEP, such as metabolism, cell signaling, cell adhesion, cell division, transport, protein synthesis and degradation were also identified.

CONCLUSIONS

Conclusion: These results demonstrate that the proteins typically found in the AEP appeared in both groups, regardless the substrate. The BISPM revealed to be a good device to be used in studies involving proteomic analysis of the AEP.

摘要

目的

获得性釉质 pellicle(AEP)是一种无细菌的有机膜,在体内形成,是唾液蛋白和糖蛋白选择性吸附到暴露于口腔环境的固体表面的结果。目的:本研究旨在使用一种新型口腔内装置(鲍鲁原位 pellicle 模型 - BISPM)比较在人和牛牙釉质上原位形成的 AEP 的蛋白质组学特征。

材料与方法

材料与方法:制备了 108 个(4×4 毫米)人和牛牙釉质样本。9 名口腔状况良好的受试者佩戴一个可摘式颌骨矫治器(BISPM),每个底物有 6 个平板随机分配。AEP 在早晨形成 120 分钟,并用浸泡在 3%柠檬酸中的电极滤纸收集。此过程重复进行三次,收集的 pellicle 进行处理以通过 LC-ESI-MS/MS 分析。将获得的质谱 MS/MS 谱图与人蛋白质数据库(SWISS-PROT)进行比对。

结果

结果:使用 BISPM 能够收集到足够量的蛋白质用于适当分析。从底物收集的 AEP 中总共发现了 51 种蛋白质。其中,15 种在两组中都有,14 种仅存在于牛牙釉质中,并 22 种仅存在于人类牙釉质中。鉴定出了通常在 AEP 中发现的蛋白质,如组蛋白 -1、免疫球蛋白α-1、免疫球蛋白α2、溶菌酶 C、statherin 和颌下腺雄激素调节蛋白 3B。还鉴定出了先前未在 AEP 中描述的蛋白质,如代谢、细胞信号传导、细胞粘附、细胞分裂、转运、蛋白质合成和降解相关的蛋白质。

结论

结论:这些结果表明,无论底物如何,通常在 AEP 中发现的蛋白质在两组中均出现。BISPM 被证明是一种用于涉及 AEP 蛋白质组学分析研究的良好装置。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dfd9/6296283/8fb843dacffe/1678-7765-jaos-27-e20180113-gf01.jpg

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