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实验性诱导牙骨质附着下的全蛋白质组分析

Global proteome profiling of dental cementum under experimentally-induced apposition.

作者信息

Salmon Cristiane R, Giorgetti Ana Paula O, Paes Leme Adriana Franco, Domingues Romênia R, Sallum Enilson Antonio, Alves Marcelo C, Kolli Tamara N, Foster Brian L, Nociti Francisco H

机构信息

Department of Prosthodontics and Periodontics, Division of Periodontics, Piracicaba Dental School, State University of Campinas, São Paulo, Brazil.

National Biosciences Laboratory, Brazilian Synchrotron Light Laboratory, Campinas, SP, Brazil.

出版信息

J Proteomics. 2016 Jun 1;141:12-23. doi: 10.1016/j.jprot.2016.03.036. Epub 2016 Apr 16.

Abstract

UNLABELLED

Dental cementum (DC) covers the tooth root and has important functions in tooth attachment and position. DC can be lost to disease, and regeneration is currently unpredictable due to limited understanding of DC formation. This study used a model of experimentally-induced apposition (EIA) in mice to identify proteins associated with new DC formation. Mandibular first molars were induced to super-erupt for 6 and 21days after extracting opposing maxillary molars. Decalcified and formalin-fixed paraffin-embedded mandible sections were prepared for laser capture microdissection. Microdissected protein extracts were analyzed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), and the data submitted to repeated measure ANOVA test (RM-ANOVA, alpha=5%). A total of 519 proteins were identified, with 97 (18.6%) proteins found exclusively in EIA sites and 50 (9.6%) proteins exclusively expressed in control sites. Fifty six (10.7%) proteins were differentially regulated by RM-ANOVA (p<0.05), with 24 regulated by the exclusive effect of EIA (12 proteins) or the interaction between EIA and time (12 proteins), including serpin 1a, procollagen C-endopeptidase enhancer, tenascin X (TNX), and asporin (ASPN). In conclusion, proteomic analysis demonstrated significantly altered protein profile in DC under EIA, providing new insights on DC biology and potential candidates for tissue engineering applications.

SIGNIFICANCE

Dental cementum (DC) is a mineralized tissue that covers the tooth root surface and has important functions in tooth attachment and position. DC and other periodontal tissues can be lost to disease, and regeneration is currently unpredictable due to lack of understanding of DC formation. This study used a model of experimentally-induced apposition (EIA) in mice to promote new cementum formation, followed by laser capture microdissection (LCM) and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) proteomic analysis. This approach identified proteins associated with new cementum formation that may be targets for promoting cementum regeneration.

摘要

未标记

牙骨质(DC)覆盖牙根,在牙齿附着和定位中具有重要功能。DC会因疾病而丧失,由于对DC形成的了解有限,目前其再生情况难以预测。本研究使用小鼠实验性诱导附着(EIA)模型来鉴定与新DC形成相关的蛋白质。拔除上颌对侧磨牙后,诱导下颌第一磨牙超萌出6天和21天。制备脱钙并经福尔马林固定的石蜡包埋下颌骨切片用于激光捕获显微切割。对显微切割得到的蛋白质提取物进行液相色谱-串联质谱分析(LC-MS/MS),并将数据提交至重复测量方差分析检验(RM-ANOVA,α = 5%)。共鉴定出519种蛋白质,其中97种(18.6%)蛋白质仅在EIA部位发现,50种(9.6%)蛋白质仅在对照部位表达。56种(10.7%)蛋白质经RM-ANOVA差异调节(p<0.05),其中24种受EIA的单独作用(12种蛋白质)或EIA与时间的相互作用(12种蛋白质)调节,包括丝氨酸蛋白酶抑制剂1a、原胶原C端肽酶增强剂、腱生蛋白X(TNX)和阿朴脂蛋白(ASPN)。总之,蛋白质组学分析表明EIA条件下DC中的蛋白质谱有显著改变,为DC生物学提供了新见解,并为组织工程应用提供了潜在候选物。

意义

牙骨质(DC)是一种覆盖牙根表面的矿化组织,在牙齿附着和定位中具有重要功能。DC和其他牙周组织会因疾病而丧失,由于对DC形成缺乏了解,目前其再生情况难以预测。本研究使用小鼠实验性诱导附着(EIA)模型促进新牙骨质形成,随后进行激光捕获显微切割(LCM)和液相色谱-串联质谱(LC-MS/MS)蛋白质组学分析。该方法鉴定出与新牙骨质形成相关的蛋白质,这些蛋白质可能是促进牙骨质再生的靶点。

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