Global proteome profiling of dental cementum under experimentally-induced apposition.
Global proteome profiling of dental cementum under experimentally-induced apposition.
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实验诱导并置下牙骨质的整体蛋白质组分析。
DOI：
10.1016/j.jprot.2016.03.036
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发表时间：
2016
影响因子：
3.3
通讯作者：
NocitiJr,FranciscoH
中科院分区：
文献类型：
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作者：
Salmon,CristianeR;Giorgetti,AnaPaulaO;PaesLeme,AdrianaFranco;Domingues,RomêniaR;Sallum,EnilsonAntonio;Alves,MarceloC;Kolli,TamaraN;Foster,BrianL;NocitiJr,FranciscoH
牙骨质（Dental cementum，DC）覆盖在牙齿的根部，对牙齿的附着和定位具有重要作用. DC可能会因疾病而丢失，由于对DC形成的了解有限，目前再生是不可预测的。本研究使用小鼠实验诱导沉积（EIA）模型来鉴定与新DC形成相关的蛋白质。拔除对侧上颌磨牙后，诱导下颌第一磨牙超萌6天和21天。制备脱钙和福尔马林固定石蜡包埋的下颌骨切片用于激光捕获显微切割。通过液相色谱-串联质谱法（LC-MS/MS）分析显微切割的蛋白质提取物，并将数据提交给重复测量ANOVA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位点发现，50种（9.6%）蛋白质仅在对照位点表达。56个（10.7%）蛋白质被RM-ANOVA差异调节（p < 0.05），其中24个蛋白质由EIA的排他性效应（12个蛋白质）或EIA和时间之间的相互作用（12个蛋白质）调节，包括serpin 1a、前胶原C-内肽酶增强子、腱生蛋白X（TNX）和asporin（ASPN）。总之，蛋白质组学分析表明，显着改变的蛋白质谱在DC下EIA，提供了新的见解DC生物学和潜在的候选人为组织工程application.SignificanceDental牙骨质（DC）是一种矿化组织，覆盖在牙根表面，并在牙齿附着和位置的重要功能。DC和其他牙周组织可能会因疾病而丢失，由于缺乏对DC形成的了解，目前再生是不可预测的。本研究采用小鼠实验诱导沉积（EIA）模型促进新牙骨质形成，然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。这种方法鉴定了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。
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 21 days 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.SignificanceDental 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.