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Histochemical studies on the distribution of calcium in the cells related to the formation of dental and periodontal hard tissues.

对与牙齿和牙周硬组织形成相关的细胞中钙分布的组织化学研究。

基本信息

批准号：
63570847
负责人：
TAKANO Yoshiro
金额：
$ 1.34万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63570847/
关键词：
ENAMEL ORGAN RAT INCISOR ENAMEL CALCIUM HISTOCHEMISTRY RAPID FREEZING FREEZE-SUBSTITUTION 急速凍結凍結置換

项目摘要

The distribution of calcium in the enamel organ, pulp and periodontal tissues was examined by means of histochemical, electron microscopic and X-ray microanalytical methods. All specimens were rapidly frozen/freeze-substituted and embedded in epoxy resin for the preservation of morphology and cell calcium. This method allowed a histochemical demonstration of cell calcium as diffuse or discrete red Ca-GBHA precipitates under the light microscope, and an electron microscopic examination of adjacent ultrathin sections.Our study revealed intense calcium reactions in the cytoplasm of both secretory and maturation ameloblasts, and its absence in other enamel organ cells. Electron microscopy and X-ray microanalytical studies confirmed the presence of calcium-containing granules and crystalline figures in the tubulovesicular structures of secretory ameloblasts, implicating the contribution of such organelles in transcellular translocation of calcium. In the pulp, Ca-GBHA reactins were shown in differentiated odontoblasts but not in other pulp cells. In molar tooth germs at the stage of root formation, Ca-GBHA reactions appeared in cementoblasts and precementoblasts located at disrupted portions of unreactive epithelial root sheath. Osteoblasts and osteoclasts on the forming and absorbing surfaces of the alveolar bone also showed intense Ca-GBHA reactions in the cytoplasm.In this project, we thus established a tissue preparation method whereby both cell morphology and unstable cell calcium is preserved. A histochemical calcium staining method was also established. With these methods, data suggesting the exclusive localization of high concentrations of calcium in cells directly related to formation or absorption of enamel, dentin or bone were presented. The elucidation of the physiological significance of present findings awaits further studies.

应用组织化学、电镜和X线微区分析方法，观察了钙在成釉器、牙髓和牙周组织中的分布。所有标本均快速冷冻/冷冻置换，并包埋在环氧树脂中以保存形态和细胞钙。这种方法允许在光镜下弥散或离散的红色Ca-GBHA沉淀的细胞钙的组织化学示范，并在相邻的ESTA切片的电子显微镜检查，我们的研究揭示了强烈的钙反应在分泌和成熟的成釉细胞的细胞质中，它的情况下，在其他成釉器官细胞。电子显微镜和X射线微量分析研究证实，分泌型成釉细胞的管状泡状结构中存在含钙颗粒和晶体结构，暗示这些细胞器在钙的跨细胞转运中的贡献。在牙髓中，Ca-GBHA反应在分化的成牙本质细胞中显示，而在其他牙髓细胞中不显示。在牙根形成阶段的磨牙牙胚中，Ca-GBHA反应出现在位于无反应的上皮根鞘破坏部分的成牙骨质细胞和前成牙骨质细胞中。牙槽骨形成面和吸收面上的成骨细胞和破骨细胞也显示出胞浆中强烈的Ca-GBHA反应。因此，在本项目中，我们建立了一种组织制备方法，该方法既保留了细胞形态，又保留了不稳定的细胞钙。同时建立了组织化学钙染色法。用这些方法，数据表明专门定位的高浓度的钙直接相关的细胞形成或吸收的釉质，牙本质或骨。本研究结果的生理意义有待进一步研究。