Research of Osteoclastogenesis in Priodontium induced by mechanical stress and its Clinical Application

机械应力诱导原牙体破骨细胞生成的研究及其临床应用

• 批准号: 12671984
• 负责人: MIKI Mirei
• 金额: $ 2.18万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671984/
• 关键词: Osteoclasts; Peiodontium; Bone resorption; Periodontal ligament; Mechanical stress; Orthodontic tooth movement; RANKL; OPG; 破骨細胞; 歯周組織; 骨吸収; 歯根膜細胞; メカニカルストレス; 歯科矯正学的歯の移動; メカニカルストレス受容; Osteoprotegerin

The periodontal tissue that is subject to mechanical loading during occlusion and mastication. The purpose of this study are to clarify the effects of mechanical stress on the osteoclastogenesis induced at a local site of stress, such as orthodontic tooth movement, and to search for there clinical application. Following results were obtained.1. The first molar of rat were moved by fixed orthodontic wire experimentally. lmmunohistochemical staining revealed that osteoprotegerin (OPG) protein accumulated adjacent to bone resorption area including osteoclasts and resorption lacunae at the compressive site of periodontal ligament. Furthermore, receptor activator of NF kappa B ligand (RANKL) protein was expressed strongly on the fibroblasts of compressive cite.2. As progenitors of osteoclasts, peripheral blood mononuclear cells (PBMCs) and human promyelocytic leukemia cell line, HL-60 were cultured directly with PDL cells. Formed multinucleated cells had osteoclastic characteristics and res … More orptive activity. Furthermore, PDL cells expressed both RANKL and OPG mRNA.3. Fifteen clones derived from periodontal ligament of SV40 tsA58 transgenic rat were established, and compared the characteristics of these cell lines by RT-PCR. RANKL and OPG were seen in 0 and 8 clones respectively and expression were changed in 3 and 0 clones respectively afler 1,25α(0H)_2D_3 (10^<-8>M) treatment.4. PDL cells under mechanical stress up-regulated osteoclastogenesis from PBMCS. Furthermore, the expression of RANKL mRNA and protein in PDL cells increased with compressive force. Cyclooxygenase 2 mRNA expression and prostaglandin E_2 production were also induced, and indomethacin inhibited the RANKL up-regulation resulting from compressive force. OPG expression remained constant.5. The number of TRAP-positive multinucleated cells cultured in the presence of conditioned media from stimulated PDL cells by tension force were reduced significantly. Tension force up-regulated OPG, RANKL and TGFβ mRNA expression in PDL cells, and increased OPG and TGFβ protein production from PDL cells. Less

咬合和咀嚼过程中承受机械负荷的牙周组织本研究的目的是阐明机械应力对局部应力部位（例如正畸牙齿移动）诱导的破骨细胞生成的影响，并探索其临床研究。获得以下结果： 1.固定正畸线移动大鼠第一磨牙，免疫组织化学染色显示骨吸收区附近有骨保护素(OPG)蛋白聚集。牙周膜受压部位的破骨细胞和吸收陷窝处的NFκB配体受体激活剂(RANKL)蛋白在受压部位的成纤维细胞上强烈表达。2．和人早幼粒细胞白血病细胞系HL-60直接与PDL细胞培养形成多核。细胞具有破骨细胞特征和抗骨活性。此外，PDL细胞均表达RANKL和OPG mRNA。建立了15个来自SV40 tsA58转基因大鼠牙周膜的克隆，并通过RT-PCR比较了这些细胞系的特征。 RANKL和OPG分别在0和8个克隆中可见，并且分别在3和0个克隆中表达变化后1，25α(0H)_2D_3(10^-8M)处理。4．机械应力下的PDL细胞上调PBMCS的破骨细胞生成。此外，PDL细胞中RANKL mRNA和蛋白质的表达随着压力而增加。 2 mRNA 表达和前列腺素 E_2 产生也被诱导，并且吲哚美辛抑制由 OPG 表达引起的 RANKL 上调。 5.在条件培养基存在下，张力刺激的PDL细胞培养的TRAP阳性多核细胞的数量显着减少。张力上调PDL细胞中的OPG、RANKL和TGFβmRNA表达，并增加OPG和PDL 细胞产生的 TGFβ 蛋白较少。

项目成果

H.KANZAKI: "RANKL Upregulation in the Compressed Area of Periodontal Ligament induces Orthodontic Tooth Movement"Journal of Dental Research. 81. A-135 (2002)

H.KANZAKI：“牙周膜受压区域的 RANKL 上调导致正畸牙齿移动”《牙科研究杂志》。

Tsuchiya, S., Chiba, M., Kishimoto, K., Nakamura, H., Mitani, H.: "Gene transfer into periodontal tissue by in vivo electroporation"Journal of Dental Research. 81. A-452 (2002)

Tsuchiya, S.、Chiba, M.、Kishimoto, K.、Nakamura, H.、Mitani, H.：“通过体内电穿孔将基因转移到牙周组织”牙科研究杂志。

Chiba, M.: "Molecular biological review on bone remodeling based on mechanical stress."the Quintessence. 21. 931-938 (2002)

Chiba, M.：“基于机械应力的骨重塑的分子生物学评论。”精髓。

C.MAISUMOTO: "Synergistic human Osteoclast differentiation inducing by the combined use of 1.25(OH)2D 3, RANKL. and M-CSF"Journal of Dental Research. 81. A105 (2002)

C.MAISUMOTO：“联合使用 1.25(OH)2D 3、RANKL. 和 M-CSF 诱导人类破骨细胞分化”《牙科研究杂志》。

千葉美麗: "メカニカルストレスによる歯根膜細胞の初期応答"日本機械学会[No.00-35]第13回バイオエンジニアリング講演会講演論文集. 192-193 (2001)

Mirei Chiba：“机械应力引起的牙周膜细胞的初始反应”第13届生物工程会议论文集，日本机械工程师学会[No.00-35]（2001年）。