Molecular biological study on the physiologic function of periodntal ligament of human deciduous tooth

人乳牙牙膜生理功能的分子生物学研究

• 批准号: 14571971
• 负责人: KIMOTO Shigenari
• 金额: $ 1.86万
• 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: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571971/
• 关键词: periodontal Ligament of deciduous teeth; fibroblast; osteoclast; vitamin D3; RANKL; osteoprotegerin; M-CSF; vitamin D_3; 破歯細胞

The expression of mRNA of osteoprotegerin(OPG), receptor activator nuclear kappa B ligand(RANKL) and macrophage colony stimulating factor(M-CSF) was investigated by RT-PCR on the cultured cells derived from periodontal ligament of human deciduous teeth(HPLF-D) and permanent teeth(HPLF-P), alveolar bone process(hOB) and gingival fibroblast(hGF). All of the cultured cells express mRNA of OPG at almost same level. The expression of M-CSF mRNA was also observed on all types of the cells. The mRNA level of secreted form of M-CSF was dominant compared to membrane-bound form. On the other hand, the expression of RANKL message was observed on HPLF-D, HPLF-P and hOB, excepted on hGF. However, the level of RANKL mRNA on HPLF-D was almost same level of hOB and it was higher than that on HPLF-P. The level of RANKL message was increased by the addition of 1 α,25 dihydroxyvitamin D_3(D_3) in dose dependent manner.These data suggested as follows1)The cells in periodontal tissues express a considerable level of mRNA of M-CSF and OPG, those are able to regulate osteoclast differentiation.2)RANKL, which is produced by osteoblasts or HPLF, is the most important factor on osteoclast differentiation in periodontal tissues.3)The level of RANKL mRNA increased by the factors playing important roll on bone metabolism (for example, D_3) is able to induce the differentiation of osteoclasts (or odontoclasts).4)Bone resorbing factors affect the cells in periodontal ligament of deciduous tooth on inducing the differentiation of osteoclst (or odontoclast), and the effect is greater than that on periodontal ligament of permanent tooth.

采用RT-PCR方法检测人乳牙牙周膜细胞（HPLF-D）、恒牙牙周膜细胞（HPLF-P）、牙槽骨细胞（hOB）和牙龈成纤维细胞（HGF）中骨保护素（OPG）、核受体活化因子κ B配体（RANKL）和巨噬细胞集落刺激因子（M-CSF）mRNA的表达。所有培养的细胞表达OPG的mRNA水平几乎相同。M-CSFmRNA在所有类型的细胞上也观察到表达。分泌型M-CSF的mRNA水平明显高于膜结合型M-CSF。另一方面，RANKL信使在HPLF-D、HPLF-P和hOB上观察到表达，但在HGF上观察到表达。HPLF-D组RANKL mRNA表达水平与hOB组基本相同，但高于HPLF-P组，1 α，25-二羟维生素D_3（D_3）可剂量依赖性地增加RANKL mRNA的表达，提示：1）牙周组织细胞表达M-CSF和OPG的mRNA，RANKL是牙周组织中破骨细胞分化的最重要的因子，由成骨细胞或HPLF产生; 3）RANKL mRNA的表达水平在骨代谢中起重要作用（如D_3）能诱导破骨细胞（或破牙细胞）的分化。4）骨吸收因子对乳牙牙周膜细胞诱导破骨细胞（或破牙细胞）分化的影响大于恒牙牙周膜细胞。