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Control of expression of dentin phosphophoryn by cellular environment.

细胞环境对牙本质磷酸化表达的控制。

基本信息

批准号：
10671730
负责人：
FUJISAWA Ryuichi
金额：
$ 1.92万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671730/
关键词：
odontoblast dentin phosphorotein dentin sialoprotein cell attachment pulp cell

项目摘要

1) Establishment of a culture system for dental pulp cellsWe developed a culture system for differentiation of dental pulp cells into odontoblast-like cells. Pulp cells were prepared from pulps of rat incisors by protease treatment. The cells were cultured in a medium containing ascorbic acid, dexamethasone and 2-glycerophosphate. The cells formed cellular nodules and then formed calcified deposits in the nodules. The cells had a high alkaline phosphatase activity. Expression of phosphophoryn/DSP, a protein specific to odontoblasts, was detected by using RT-PCR. Expression of BGP, a protein specific to calcified tissues, was also detected. These phenotypes indicates that the cell were differentiated into odontoblast-like cells.2) Effects of bioactive factors on the expression of phosphophoryn/DSPBy using the above culture system, we examined effects of several reagents on the expression of odontoblast-specific genes. Dexamethasone, a differentiation factor for osteoblasts, enhanced the expression of the phosphophoryn/DSP. This agent promoted the differentiation of the pulp cells. Another osteotropic hormone, 1,25(OH)ィイD22ィエD2DィイD23ィエD2 had no effect on the expression. Bis-phosphonate, an agent inhibitory to calcification, activated the expression of the phosphophoryn/DSP, although it reduced the calcification. This result indicates that the bis-phosphonate promoted the differentiation of the pulp cells. Inhibition of the calcification can be caused by its physico-chemical activity.

1)牙髓细胞培养体系的建立我们建立了牙髓细胞向成牙本质细胞样细胞分化的培养体系。采用蛋白酶处理大鼠切牙牙髓，制备牙髓细胞。将细胞培养在含有抗坏血酸、地塞米松和2-甘油磷酸盐的培养基中。细胞形成细胞结节，然后在结节中形成钙化沉积物。细胞具有高碱性磷酸酶活性。RT-PCR检测成牙本质细胞特异性蛋白磷酸化蛋白（phosphophoryn/DSP）的表达。还检测了钙化组织特异性蛋白BGP的表达。这些表型表明细胞已分化为成牙本质细胞样细胞。2）生物活性因子对磷酸化蛋白/DSP表达的影响利用上述培养体系，我们检测了几种试剂对成牙本质细胞特异性基因表达的影响。地塞米松，成骨细胞的分化因子，增强磷酸化/DSP的表达。该试剂促进牙髓细胞的分化。另一种促骨激素1，25（OH）β D22 β D2 D β D23 β D2对表达无影响。双膦酸盐，一种抑制钙化的药物，激活了磷酸蛋白/DSP的表达，尽管它减少了钙化。这一结果表明，双膦酸盐促进了牙髓细胞的分化。抑制钙化可能是由于其物理化学活性。