| Project/Area Number |
13671985
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
病態科学系歯学(含放射線系歯学)
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| Research Institution | Nihon University |
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Principal Investigator |
MAENO Masao Nihon University, School of Dentistry, Department of Oral Health Sciences, Professor, 歯学部, 教授 (60147618)
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| Co-Investigator(Kenkyū-buntansha) |
SUZUKI Naoto Nihon University, School of Dentistry, Department of Biochemistry, Assistant Professor, 歯学部, 講師 (10226532)
ITO Koichi Nihon University, School of Dentistry, Department of Periodontology, Professor, 歯学部, 教授 (90102607)
OTSUKA Kichibee Nihon University, School of Dentistry, Department of Biochemistry, Professor, 歯学部, 教授 (50059995)
YAMAGUCHI Yoko Nihon University, School of Dentistry, Department of Biochemistry, Assistant, 歯学部, 副手 (00239922)
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2003: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2002: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2001: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | 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 zinc / carnosine / bone morphogenetic proteins / 未分化間葉系細胞 / 筋芽細胞 / オステオカルシン / アルカリホスファターゼ / enamel matrix-derived protein / ヒト歯根膜細胞 / bone sialoprotein / integrin αvβ5 / RGD配列ペプチド / osteopontin |
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| Research Abstract |
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
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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
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