Cell biological study on physiological root resorption of deciduous teeth ; potentiality to support odontoclast differentiation on periodontal ligament fibroblasts derived from of human deciduous teeth

乳牙生理性牙根吸收的细胞生物学研究；

• 批准号: 10671957
• 负责人: KIMOTO Shigenari
• 金额: $ 1.98万
• 依托单位: Kanagawa Dental College
• 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 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671957/
• 关键词: deciduous tooth; periodontal ligament; fibroblast; odontoclast; osteoclast; (1)乳歯; (2)歯根膜; (3)線維芽細胞; (4)破歯細胞; 生理的歯根吸収

It was suggested that the fibroblasts derived from periodontal ligament of human permanent teeth (HPLF) and deciduous teeth (HPLF-Y) inhibit the formation of odontoclast (or osteoclast)-like cells using coculture system with mouse bone marrow cells (BMC), while the osteoblasts derived from human alveolar bone (hOB) has no effect on that compared to the culture of BMC alone. The inhibitory effects on the osteoclast-like cells (OC) formation in contact with HPLF or HPLF-Y were more significant than the coculture at non-contact condition between BMC and HPLF or HPLF-Y.On the other hand, pretreated HPLF and HPLF-Y with 1 α 25(OH)_2 vitaminD_3 (D_3) and dexamethasone (Dex) did not inhibited tartrate- resistant acid phosphatase (TRAP) positive multinucleated cells (MNC) formation in the coculture system. Then we performed RT-PCR to examine the mRNA level for macrophage colony stimulating factor (M-CSF), one of the essential factors for the differentiation of osteoclast precursor in early stage, on total RNA isolated from HPLF-Y, hOB and the fibroblasts derived from human gingival tissue (hGF) treated with D_3, Dex or both of them. As the result of semi-quantitative RT-PCR, the expression of mRNA for secreted form M-CSF (M-CSFs) was predominant over that for membrane-bound form M-CSF (M-CSFm) on hOB and hGF, however mRNA expression of both forms was same level on HPLF- Y.Moreover, the level of mRNA for M-CSFm increased by D_3 treatment and the effect was more remarkable in the presence of Dex while mRNA expression for M-CSFs was not affected by D_3 and Dex treatment. On the other hand, Dex treatment increased the ratio of M-CSFm mRNA level in total expression of both M-CSFs and M-CSFm on HPLF-Y and hGF, while it was reduced by D_3 treatment. These results suggest that the proportion of mRNA level of M-CSFm to that of M-CSFs on the fibroblast is different from osteoblast and that the ratio is altered by soluble factors related to bone metabolism such as D_3 and Dex.

结果表明，人恒牙和乳牙牙周韧带成纤维细胞与小鼠骨髓细胞（BMC）共培养可抑制破牙细胞样细胞的形成，而人牙槽骨（hOB）成骨细胞与单独培养BMC相比，对破牙细胞样细胞的形成无明显影响。BMC与HPLF或HPLF- y接触时对破骨细胞样细胞（OC）形成的抑制作用比BMC与HPLF或HPLF- y非接触共培养更显著。另一方面，1 α 25(OH)_2维生素ind_3 （D_3）和地塞米松（Dex）预处理的HPLF和HPLF- y在共培养体系中没有抑制酒石酸盐抗性酸性磷酸酶（TRAP）阳性多核细胞（MNC）的形成。然后我们采用RT-PCR检测巨噬细胞集落刺激因子（M-CSF）的mRNA水平，巨噬细胞集落刺激因子（M-CSF）是早期破骨细胞前体分化的重要因子之一，对HPLF-Y、hOB和来源于人牙龈组织的成纤维细胞（hGF）的总RNA进行检测。半定量RT-PCR结果显示，分泌型M-CSF (M-CSF) mRNA在hOB和hGF上的表达高于膜结合型M-CSF (M-CSFm) mRNA，而两种形式的mRNA在HPLF- y上的表达水平相同，且D_3处理使M-CSFm mRNA水平升高，且在Dex存在时效果更为显著，而D_3和Dex处理对M-CSF mRNA表达没有影响。另一方面，Dex处理增加了M-CSFm mRNA水平在HPLF-Y和hGF上M-CSFm和M-CSFm总表达量中的比例，而D_3处理则降低了M-CSFm mRNA水平。这些结果表明，成纤维细胞上M-CSFm与M-CSFs mRNA水平的比例与成骨细胞不同，这一比例受与骨代谢相关的可溶性因子如D_3和Dex的影响。

项目成果

S.Kimoto, M.Matsuzawa, S.Matsubara, T.Komatsu, N.Uchimura, T.Kawase, S.Saito: "Cytokine secretion of periodontal ligament fibroblasts derived from human deciduous teeth : effect of mechanical stress on the secretion of transforming growth factor-b1 and ma

S.Kimoto、M.Matsuzawa、S.Matsubara、T.Komatsu、N.Uchimura、T.Kawase、S.Saito：“人乳牙牙周膜成纤维细胞的细胞因子分泌：机械应力对转化细胞分泌的影响

S.Kimoto,M.Matsuzawa,S.Matsubara,T.Komatsu,N.Uchimura,T.Kawase,S.Saito: "Cytokine secretion of periodontal ligament fibroblasts derived from human deciduous teeth:effet of mechanical stress on the secretion of transforming growth factor-β1 and macrophage

S.Kimoto，M.Matsuzawa，S.Matsubara，T.Komatsu，N.Uchimura，T.Kawase，S.Saito：“人乳牙牙周膜成纤维细胞的细胞因子分泌：机械应力对转化细胞分泌的影响生长因子-β1和巨噬细胞