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The study of the factors of root development and reparative dentine formation

牙根发育及修复性牙本质形成因素的研究

基本信息

批准号：
18592098
负责人：
MASUDA Yoshiko
金额：
$ 2.59万
依托单位：
Showa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18592098/
关键词：
reparative dentine odontoblast-like cell proteozlycan

项目摘要

We prepared the cDNA library as odontoblast-like cell enriched cDNA in vivo with mRNA from the laser irradiated pulp of a rat incisor. The glypican-1 (GPC-1) was detected among the identical genes in the cDNA library as odontoblast-like cell enriched cDNA in vivo. GPC-1 is a member of a family of glycosylphosphatidylinositol anchored cell surface heparin sulfate proteoglycans (HSPG) .Objectives : The purpose of this study was to identify the expression of HSPG in odontoblast-like cells and evaluate their relation to the formation of reparative dentine.Methods: 6 and 10 days after Nd: YAG laser irradiation in the pulp, mandibles of rats were dissected and histological procedure was proceeded. We detected the expression of the GPC-1 and Syndecan-1 inununohistologically. To investigate the expression of the GPC-1 gene, Syndecan-1, -2, -3, -4 gene comparing with Tgf-beta 1, osteocalcin, Dspp, we used the cell culture system that reflects the dentine formation in dental pulp tissue. The mandibles were removed from seven male wistar rats (5 weeks old) from the same litter. The pulp tissues were cut into several pieces and incubated in a sterile enzyme solution containing 0.1 % collagenase, 0.05% trypsin. The total RNA were extracted from the cultured cells on days 3, 6, 10 and 15. The RT-PCR amplification was performed.Results : The immunohistological examination showed stainings of GPC-1 and Syndecan-1 were observed in odontoblast-like cells. In the cell culture system, the expression of mRNA of GPC-1, Syndecan-land Syndecan-4 gradually increased by 15 days, but the expression of Syndecan-2 and Syndecan-3 were almost same during 15 days. The levels of relative quantity mRNA of GPC-1, Syndecan-land syndecan-4 were increased with the process of the odontoblast-like cell differentiation.Conclusion : These findings suggest that GPC-1, Syndecan-land synclecan-4 might be associated with differentiation of odontoblast-like cell.

我们用激光照射大鼠门牙牙髓的mRNA在体内制备成牙母细胞样细胞富集cDNA文库。glypican-1 （GPC-1）在体内作为牙髓细胞样细胞富集的cDNA在cDNA文库中被检测到。GPC-1是糖基磷脂酰肌醇锚定细胞表面硫酸肝素蛋白聚糖（HSPG）家族的成员。目的：本研究旨在鉴定HSPG在成牙细胞样细胞中的表达，并评价其与修复性牙本质形成的关系。方法：在Nd: YAG激光照射牙髓后6、10 d解剖大鼠下颌骨并进行组织学检查。病理检测GPC-1和Syndecan-1的表达。为了比较GPC-1基因、Syndecan-1、-2、-3、-4基因与tgf - β 1、骨钙素、Dspp基因的表达，我们采用反映牙髓组织牙本质形成的细胞培养系统。取同一窝产的7只5周龄雄性wistar大鼠下颌骨。将牙髓组织切成几块，在含有0.1%胶原酶、0.05%胰蛋白酶的无菌酶溶液中培养。分别于第3、6、10、15天提取总RNA。进行RT-PCR扩增。结果：免疫组织学检查显示成牙细胞样细胞GPC-1和Syndecan-1染色。在细胞培养体系中，GPC-1、Syndecan-4和Syndecan-4 mRNA的表达量在15 d内逐渐升高，而Syndecan-2和Syndecan-3 mRNA的表达量在15 d内基本持平。GPC-1、syndecan-和syndecan-4 mRNA的相对表达量随着成牙细胞样细胞的分化而升高。结论：GPC-1、Syndecan-land synclecan-4可能与成牙细胞样细胞的分化有关。