The elucidation of action mechanism of enamel matrix derivative on the reconstruction of alveolar bone.

釉质基质衍生物对牙槽骨重建作用机制的阐明。

• 批准号: 13671985
• 负责人: MAENO Masao
• 金额: $ 2.37万
• 依托单位: Nihon University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671985/
• 关键词: Enamel matrix derivative; Periodontal ligament cells; Cell attachment; Cell differentiation; C2C12 Cells; β-alanyl-L-hisitidinato zinc; Carnosine; Bone morphogenetic proteins; enamel matrix derivative; ヒト歯根膜由来細胞; 骨芽細胞; β-alanyl-L-histidinato zi

Although enamel matrix derivative (EMD) can stimulate attachment of human periodontal ligament (HPDL) cells to the root surface, the biological mechanism of this phenomenon is unclear. Therefore, we determined which molecules in EMD are involved in the attachment of HPDL cells, and which types of integrins on the cell surface mediatethe interaction between the cells and EMD. Our results suggest that the attachment of HPDL cells to EMD can be mediated by interaction between a bone sialoprotein-like molecule and integrin αvβ3 on the cell surface (J Periodontol 2001;72:1520-1526).Although EMD initiate de novo cementum and bone formation by stimulating and inducing differentiation of mesenchymal cells in the periodontal ligament, the molecular mechanism of this phenomenon is not fully understood. Therefore, we determined the effect of EMD on the differentiation of pluripotential mesenchymal cells(C2C12 cells). C2C12 cells cultured in differentiation medium without EMD altered their phenoty … More pe to myoblasts, exhibiting positive reactions to desmin and myosin heavy chains by immunological analysis. However, the cells cultured in the presence of EMD were strongly inhibited from developing into myoblasts, and showed high alkaline phosphatase (ALPase) avtivity that was approximately 2 to 4 times greater than that of the vehicle. The mRNA expression of ALPase, osteocalcin, and type X collagen was increased markedly by the EMD-stimulated medium, whereas the expression of desmin, MyoD, and lipoprotein lipase was drastically decreased. These results suggest that. EMD converts the differentiation pathway of C2C12 cells into the osteoblast and/or chondroblast lineage (J Periodontol 2002;73:543.550).We determined the effect of β-alanyl-L-histidinato zinc (AHZ) and β-alanyl-L-histidine (carnosine) on the differentiation of HPDL cells. RUNX2/Cbfa1 expression increased markedly in cells cultured with AHZ, while Sox9 expression increased slightly. By contrast, RUNX2/Cbfa1 and Sox9 expressions increased markedly with carnosine. Bone morphogenetic protein-2 (BMP-2) and BMP-7 expressions was much higher than that of controls in cultures with AHZ and carnosine. The expression of BMP receptors increased markedly in cells cultured with AHZand carnosine. The phosphorylation of Smad1, a signal-transducing molecule for BMP-2 and BMP-7, was increased markedly in cultures with AHZ and camosine. These results suggest that AHZ and carnosine divert the differentiation pathway of HPDL cells into the osteoblast and/or chondroblast line age via the autocrine action of BMP-2 or BMP-7 produced by the cells (Dent Jpn 2003;39:114-118, Life Sciences 2004;74:2493-2504, J Periodontol Res 2004;39:in press) Less

尽管釉质基质衍生物(Emd)可以促进人牙周膜(HPDL)细胞在牙根表面的附着，但其生物学机制尚不清楚。因此，我们确定了EMD中哪些分子参与了HPDL细胞的附着，以及细胞表面哪些类型的整合素介导了细胞与EMD之间的相互作用。我们的结果表明，HPDL细胞与牙周膜间充质细胞的黏附可以通过骨涎蛋白样分子与细胞表面整合素αvβ3之间的相互作用来介导(J Perodontol 2001；72：1520-1526)。虽然骨膜间充质细胞通过刺激和诱导牙周膜间充质细胞的分化来启动牙骨质和骨的形成，但这种现象的分子机制尚不完全清楚。因此，我们确定了EMD对多潜能间充质细胞(C2C12细胞)分化的影响。分化培养的C2C12细胞表型…发生改变免疫分析显示结蛋白和肌球蛋白重链呈阳性反应。然而，在EMD的存在下培养的细胞被强烈地抑制向成肌细胞的发育，并且表现出高的碱性磷酸酶(ALPase)活性，大约是载体的2-4倍。EMD刺激后，ALPase、骨钙素和X型胶原的mRNA表达显著增加，而结蛋白、MyoD和脂蛋白脂肪酶的表达显著降低。这些结果表明。EMD将C2C12细胞的分化途径转化为成骨细胞和/或软骨母细胞系(J Perodontol 2002；73：543.550)。我们检测了β-丙氨酰-L-组氨酸锌和β-丙氨酰-L-组氨酸(肌肽)对人牙周膜细胞分化的影响。在AHZ作用下，Runx2/Cbfa1的表达显著增加，而Sox9的表达略有增加。相反，RUNX2/Cbfa1和Sox9的表达随着肌肽的增加而显著增加。骨形态发生蛋白-2(BMP-2)和骨形态发生蛋白-7(BMP-7)在AHZ和肌肽作用下的表达明显高于对照组。在AHZ和肌肽作用下，BMP受体的表达明显增加。BMP-2和BMP-7的信号转导分子Smad1在AHZ和Camosine的作用下，其磷酸化水平显著增加。这些结果表明，AHZ和肌肽通过细胞产生的BMP-2或BMP-7的自分泌作用将HPDL细胞的分化途径转向成骨细胞和/或软骨母细胞系AGE(Dent Jpn 2003；39：114-118，Life Science 2004；74：2493-2504，J Perodontol res 2004；39：in Press)

项目成果

Masao Maeno, Emi Ito-Kato, Naoto Suzuki, Tsuyoshi Takada, Tadahiro Takayama, Koichi Ito, Kichibee Otsuka: "Effect of β-alanyl-L-histidinato zinc on the differentiation pathway of human periodontal ligament cells"Life Sciences. 74. 2493-2504 (2004)

Masao Maeno、Emi Ito-Kato、Naoto Suzuki、Tsuyoshi Takada、Tadahiro Takayama、Koichi Ito、Kichibee Otsuka：“β-丙氨酰-L-组氨酸锌对人牙周膜细胞分化途径的影响”生命科学 74. 2493。 -2504 (2004)

Mariko Ohyama, Naoto suzuki, Yoko Yamaguchi, Masao Maeno, Kichibee Otsuka, Koichi Ito: "Effect of enamel matrix derivative on the differentiation of C2C12 cells"Journal of Periodontology. Vol.73 No.5. 543-550 (2002)

Mariko Ohyama、Naoto suzuki、Yoko Yamaguchi、Masao Maeno、Kichibee Otsuka、Koichi Ito：“牙釉质基质衍生物对 C2C12 细胞分化的影响”牙周病学杂志。

Masao Maeno, Emi Ito, Naoto Suzuki, Tsuyoshi Takada, Koichi Ito, Kichibee Otsuka: "Carnosine modulates osteoblastic and chondroblastic differentiation of human periodontal ligament cells"Dentistry in Japan. 39. 114-118 (2003)

Masao Maeno、Emi Ito、Naoto Suzuki、Tsuyoshi Takada、Koichi Ito、Kichibee Otsuka：“肌肽调节人牙周膜细胞的成骨细胞和软骨细胞分化”日本牙科。

Naoto Suzuki.Mariko Ohyama, Masao Maeno, Koichi Ito, Kichibee Otsuka: "Attachment of human periodontal ligament cells to enamel matrix-derived protein is mediated via interaction between BSP-like molecules and integrin αvβ3"Journal of Periodontology. Vol.

Naoto Suzuki.Mariko Ohyama、Masao Maeno、Koichi Ito、Kichibee Otsuka：“人牙周膜细胞与牙釉质基质衍生蛋白的附着是通过 BSP 样分子和整合素 αvβ3 之间的相互作用介导的”《牙周病学杂志》卷。