Helium ion microscopy of enamel crystallites and extracellular tooth enamel matrix.
Helium ion microscopy of enamel crystallites and extracellular tooth enamel matrix.
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DOI：
10.3389/fphys.2014.00395
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发表时间：
2014
影响因子：
4
通讯作者：
Goetze B
中科院分区：
文献类型：
--
作者：
Bidlack FB;Huynh C;Marshman J;Goetze B
牙釉质研究中的一个未解决的问题是同时分析矿物和有机相在给定样本中的三维（3D）组织中的空间分辨率。本研究旨在解决这一需求，使用高分辨率成像来分析釉质基质的3D结构组织，特别是釉原蛋白，与形成釉质晶体有关。将来自野生型小鼠的化学固定的半下颌骨包埋在LR白色丙烯酸树脂中，抛光并简单蚀刻以暴露发育中的牙釉质中的有机基质。全长釉原蛋白用特异性抗体和10 nm免疫金标记。这使我们能够使用和比较两种不同的高分辨率成像技术来分析未涂层样品。氦离子显微镜（HIM）应用于研究有机和矿物结构的空间组织，而场发射扫描电子显微镜（FE-SEM）在各种模式下，包括背散射电子检测，使我们能够辨别金标记的蛋白质。在分泌晚期至成熟早期阶段的野生型釉质显示邻近成釉细胞的釉质基质蛋白（包括全长釉原蛋白）的纵向平行排列，其随后随着离矿化前沿的距离增加而转变成更异质的外观。邻近晶体束的基质形成光滑的蕾丝鞘，而在釉质棱柱之间，它被组织成散布着棒状蛋白质的球形成分。这些发现首先突出了，釉质基质的异质组织可以在矿化的整块样品中可视化。其次，我们的研究结果表明，这些技术的组合是一个强大的方法来阐明在纳米分辨率的矿化组织中的有机基质分子的三维结构组织。
An unresolved problem in tooth enamel studies has been to analyze simultaneously and with sufficient spatial resolution both mineral and organic phases in their three dimensional (3D) organization in a given specimen. This study aims to address this need using high-resolution imaging to analyze the 3D structural organization of the enamel matrix, especially amelogenin, in relation to forming enamel crystals. Chemically fixed hemi-mandibles from wild type mice were embedded in LR White acrylic resin, polished and briefly etched to expose the organic matrix in developing tooth enamel. Full-length amelogenin was labeled with specific antibodies and 10 nm immuno-gold. This allowed us to use and compare two different high-resolution imaging techniques for the analysis of uncoated samples. Helium ion microscopy (HIM) was applied to study the spatial organization of organic and mineral structures, while field emission scanning electron microscopy (FE-SEM) in various modes, including backscattered electron detection, allowed us to discern the gold-labeled proteins. Wild type enamel in late secretory to early maturation stage reveals adjacent to ameloblasts a lengthwise parallel alignment of the enamel matrix proteins, including full-length amelogenin proteins, which then transitions into a more heterogeneous appearance with increasing distance from the mineralization front. The matrix adjacent to crystal bundles forms a smooth and lacey sheath, whereas between enamel prisms it is organized into spherical components that are interspersed with rod-shaped protein. These findings highlight first, that the heterogeneous organization of the enamel matrix can be visualized in mineralized en bloc samples. Second, our results illustrate that the combination of these techniques is a powerful approach to elucidate the 3D structural organization of organic matrix molecules in mineralizing tissue in nanometer resolution.
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DOI：
10.1155/2013/684607
发表时间：
2013-09-16
期刊：
ISRN dentistry
影响因子：
--
作者：
Bartlett JD
通讯作者：
Bartlett JD
影响因子：
3
作者：
Gallon, Victoria;Chen, Lisha;Yang, Xiudong;Moradian-Oldak, Janet
通讯作者：
Moradian-Oldak, Janet
影响因子：
5.6
作者：
Lacruz, Rodrigo S.;Smith, Charles E.;Moffatt, Pierre;Chang, Eugene H.;Bromage, Timothy G.;Bringas, Pablo, Jr.;Nanci, Antonio;Baniwal, Sanjeev K.;Zabner, Joseph;Welsh, Michael J.;Kurtz, Ira;Paine, Michael L.
通讯作者：
Paine, Michael L.
影响因子：
3
作者：
Fang, Ping-An;Margolis, Henry C.;Conway, James F.;Simmer, James P.;Beniash, Elia
通讯作者：
Beniash, Elia
影响因子：
3
作者：
He, Xiaodong;Li, Wu;Habelitz, Stefan
通讯作者：
Habelitz, Stefan