MECHANISM FOR THE INHIBITION OF ODONTOCLAST FORMATION IN PULP TISSUE

抑制牙髓组织中破牙细胞形成的机制

• 批准号: 05671539
• 负责人: KASUGAI Shohei
• 金额: $ 1.41万
• 依托单位: TOKYO MEDICAL AND DENTAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05671539/
• 关键词: DENTAL PULP; ODONTOCLAST; OSTEOCLAST; CYTOKINES; サイトカイン

Our working hypothesis is the existence of the mechanism for the inhibition of odontoclast formation in pulp tissue, and it is possible that dental pulp cells produce some cytokines which inhibit odontoclast formation. First, we added the conditioned medium of dental pulp cells (RPC-C2A cells) into osteoclast-formation system using mouse bone marrow culture, and we observed the inhibition of osteoclast formation. After dialysis of the conditioned medium this inhibition was also observed, supporting our hypothesis. We repeated this experiment several times, however, stimulation of osteoclast formation was observed when we increased the amount of conditioned medium. Morever, co-culture experimant, in which dental pulp cells and bone marrow cells were separated with membrane demonstrated the stimulation of osteoclast formation. Although the results were complicated, we can conclude that dental pulp cells produce some cytokines which affect osteoclast formation. Characterization of such cytokines is remained to be investigated.On the other hand, various cytokines which affect the formation and/or activation of osteoclast have been reported. We did the preliminary experiment concerning the expression of various cytokines in human dental pulp tissues which were clinically obtained, using RT-PCR.We feel usefulness of this method and this experiment is underway. Using RNA from mouse dental pulp, we did the same experiment, and we found the expressions of TGF-beta and IL-6 but not MCSF which is necessary for osteoclast formation. Now, we are confirming our findings using Northern Blotting.

我们的工作假设是，牙髓组织中存在抑制破牙本质细胞形成的机制，牙髓细胞可能产生抑制破牙本质细胞形成的细胞因子。首先，我们将牙髓细胞条件培养液（RPC-C2 A细胞）加入小鼠骨髓培养的破骨细胞形成系统中，观察破骨细胞形成的抑制作用。透析条件培养基后，也观察到这种抑制，支持我们的假设。我们重复这个实验几次，然而，当我们增加条件培养基的量时，观察到破骨细胞形成的刺激。在牙髓细胞和骨髓细胞用膜分离的共培养实验中，证实了破骨细胞的形成。虽然实验结果比较复杂，但我们可以得出结论，牙髓细胞产生的一些细胞因子影响破骨细胞的形成。这些细胞因子的特性尚待进一步研究，而影响破骨细胞形成和/或活化的各种细胞因子已被报道。我们用RT-PCR对临床获得的人牙髓组织中各种细胞因子的表达进行了初步实验，我们认为这种方法是有用的，这项实验正在进行中。我们用小鼠牙髓RNA做了同样的实验，发现TGF-β和IL-6的表达，但没有发现破骨细胞形成所必需的MCSF。现在，我们正在用北方印迹法证实我们的发现。

项目成果

春日井 昇平: "培養ラット歯髄細胞による石灰化組織形成系の確立とこの実験系において産生される基質に関する研究" 歯科基礎医学会雑誌. 36(抄). 75 (1994)

Shohei Kasugai：“使用培养的大鼠牙髓细胞建立矿化组织形成系统以及对该实验系统中产生的基质的研究”日本基础牙科医学会杂志 36（摘要）75。

A.Soekanto 他: "Toxicity of camphorated phenol and camphorated parachlorophonol in dental pulpcell culture" Journal of Endodontics. (印刷中).

A. Soekanto 等人：“牙髓细胞培养物中樟脑酚和樟脑对氯酚的毒性”《牙髓学杂志》（正在出版）。

菊川 郁雄 他: "培養ラット歯根膜細胞の性質に関する研究" 歯科基礎医学会雑誌. 36(抄). 168 (1994)

Ikuo Kikukawa 等：“培养的大鼠牙周膜细胞特性的研究”日本基础牙科医学会杂志 36（摘要）168。

Kasugai S,Shibata S,Suzuki S,Susami T,Ogura H.: "Characterization of a system of mineralized-tissue formation by rat dental pulp cells in culture" Archives of Oral Biology. 38. 769-777 (1993)

Kasugai S、Shibata S、Suzuki S、Susami T、Ogura H.：“培养中大鼠牙髓细胞矿化组织形成系统的表征”口腔生物学档案。

Kasugai S et al.: "Characterization of a system of mineralized-tissue formation by rat dental pulp cells in culture" Archives of Oral Biology. 38. 769-777 (1993)

Kasugai S 等人：“培养中大鼠牙髓细胞矿化组织形成系统的表征”口腔生物学档案。