Effects of various factors produced by mechanically stressed human periodontal ligament fibroblasts (HPLF) on bone-related cells

机械应力下人牙周膜成纤维细胞（HPLF）产生的多种因子对骨相关细胞的影响

• 批准号: 07672215
• 负责人: SAITO Isao
• 金额: $ 1.6万
• 依托单位: Niigata University, School of Dentistry
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07672215/
• 关键词: Tooth movement; Mechanical stress; Periodontal fibroblasts; Calcitonin gene-related peptide; Alveolar osteoblasts; MC3T3-E1; Osteoclasts; ヒト歯根膜線維芽細胞; MC3T3-E1細胞; 持続的加圧刺激; アルカリフォスファターゼ活性; サイクリックAMP産生量

Various studies in vitro have shown that the physically stressed human periodontal ligament fibroblasts (HPLF) may have an important role in the regulation of bone remodeling evoked by orthodontic tooth movement. To determine whether conditioned medium (S-CM) obtained from the mechanically stressed HPLF would affect the osteoblastic function or activity, we investigated ALPase activity and cAMP production in clonal mouse osteoblast-like cells (MC3T3-E1), after an addition of S-CM.HPLF and MC3T3-E1 were routinely cultured in Dulbecco's Modified Eagle's Medium (D-MEM) containing 15% fetal bovine serum (FBS) at 37ﾟC under 5% CO_2 in air. At the confluent phase, continuously compressive stress of 1.0 g/cm^2 was applied to HPLF for 1,3,6,12 and 24 h. Each conditioned medium was gathered, and then added to MC3T3-E1. At the end of each experimental period, ALPase activity and cAMP production were measured by a modification of the method of Puzan and Brand, and the use of enzyme immunoassay kit for cAMP,respectively.The results obtained in this project were as follows :1.Addition of S-CM induced increases in both ALPase activity and cAMP production in MC3T3-E1, time-dependently, as observed in directly stressed MC3T3-E1.2.Especially in the presence of S-CM,significant increase in ALPase activity and cAMP production in MC3T3-E1 was recognized after 12h and 6h, respectively (p<0.01).3.S-CM in the presence of Indomethacin, an inhibitor of PGEs, increased neither ALPase activity nor cAMP production in MC3T3-E1.These findings suggest that several factors produced by HPLF subjected to continuously compressive stress might affect differentiation and/or cellular responses of MC3T3-E1. In addition, PGEs appear to be one of these factors and to act as a modulator of cellular activities of MC3T3-E1.

各种体外研究表明，受到物理压力的人牙周膜成纤维细胞（HPLF）可能在正畸牙齿移动引起的骨重塑的调节中发挥重要作用。为了确定从机械应激 HPLF 获得的条件培养基 (S-CM) 是否会影响成骨细胞功能或活性，我们研究了添加 S-CM 后克隆小鼠成骨细胞样细胞 (MC3T3-E1) 中 ALPase 活性和 cAMP 的产生。HPLF 和 MC3T3-E1 在含有 15% 胎儿的杜尔贝科改良伊格尔培养基 (D-MEM) 中常规培养 牛血清（FBS），37°C，5% CO_2，空气中。在汇合阶段，对 HPLF 连续施加 1.0 g/cm^2 的压应力 1、3、6、12 和 24 h。收集每种条件培养基，然后添加至MC3T3-E1。在每个实验期结束时，分别通过改进的 Puzan 和 Brand 方法以及使用 cAMP 酶联免疫分析试剂盒测量 ALPase 活性和 cAMP 产量。 本项目获得的结果如下：1. 添加 S-CM 诱导 MC3T3-E1 中 ALPase 活性和 cAMP 产量的增加，呈时间依赖性，如直接应激中观察到的那样。 MC3T3-E1.2.特别是在存在 S-CM 的情况下，12 小时和 6 小时后，MC3T3-E1 中的 ALPase 活性和 cAMP 生成分别显着增加 (p<0.01)。3.在 PGE 抑制剂吲哚美辛存在下，S-CM 既不增加 MC3T3-E1 中的 ALPase 活性，也不增加 cAMP 生成。这些发现表明，由HPLF 持续受到 压缩应力可能会影响 MC3T3-E1 的分化和/或细胞反应。此外，PGE 似乎是这些因素之一，并且充当 MC3T3-E1 细胞活性的调节剂。

项目成果

花見重幸: "実験的歯の移動時における三又神経節CGRP陽性ニューロンの反応に関する免疫組織化学的研究" 日本矯正歯科学会雑誌. 55(3). 211-222 (1996)

Shigeyuki Hanami：“实验性牙齿运动期间三叉神经节 CGRP 阳性神经元反应的免疫组织化学研究”日本正畸学会杂志 55（3）（1996）。

OCHI,Kanako: "Expression of TrKB-like immunoreactirty in non-neural cells of rat periodontal ligament" Archs oral Biol.42(in press). (1997)

OCHI，Kanako：“大鼠牙周膜非神经细胞中 TrKB 样免疫反应的表达”Archsoral Biol.42（印刷中）。

花見重幸: "実験的歯の移動時における三叉神経節CGRP陽性ニューロンの反応に関する免疫組織化学的研究" 日本矯正歯科学会雑誌. 55. 211-222 (1996)

Shigeyuki Hanami：“实验性牙齿运动期间三叉神经节中 CGRP 阳性神经元反应的免疫组织化学研究”日本正畸学会杂志 55. 211-222 (1996)。

SAITO, Isao: "The Biological Mechanisms of Tooth Movement and Craniofauol Adaptation" DAVIDOVITCH, Ze'ev, 623 (1996)

SAITO, Isao：“牙齿运动和颅骨适应的生物机制” DAVIDOVITCH, Zeev，623 (1996)

Isao SAITO: "Effects of calcitonin gene-related peptide (CGRP) on mechanically stressed osteoblasts in vivo and osteoblastic cells in vitro." In : Davidovich Z.ed. The biological mechanisms of tooth movement and craniofacialadaptation (an international (c

Isao SAITO：“降钙素基因相关肽（CGRP）对体内机械应激的成骨细胞和体外成骨细胞的影响。”