Characterization of factors controlling the differentiation of periodontal ligament cells

控制牙周膜细胞分化的因素的表征

• 批准号: 11470460
• 负责人: KASUGAI Shohei
• 金额: $ 8.64万
• 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 Scientific Research (B).
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470460/
• 关键词: Periodontal Tissue; Regeneration; Amelogenin; S100A4; Calcium Binding Proteins; Mineralization; Mechanical Stress; Bone; 歯根膜; 分化; エナメルタンパク

The purpose of this research was to characterize molecules which control the differentiation and function of periodontal ligament (PDL) cells. We constructed cDNA library of bovine PDL tissue and cloned cDNA of S100A4, one of the calcium binding proteins. We observed that mRNA expression level of S100A4 in PDL of erupted teeth was high and that PDL cells secreted this protein, which localized adjacent to collagen fibers. Addition of recombinant S100A4 protein to oseteoblastic culture inhibited mineralization. In culture, PDL cells under mechanical stress inceased mRNA expression of S100A4 together with cytoskeletal proteins. Although the expression level of this protein in MC3T3-E1 cells (osteoblastic cells) was low, inhibition of S100A4 expression in MC3T3-E1 cells stimulated mineraliztion in culture. These results strongly indicate that S100A4 is an inhibitor of meranlization in PDL tissue. On the other hand, porcine enamel matrix derivative (EMD) stimulated growth and differentiation of bovine PDL cells and Kusa cells (derived from mouse bone marrow), resulting the stimulation of mineralization in these culture. The stimulatory effects on osteoblastic cells could also contribute to periodontal tissue regeneration. It is important to clarify effective molecule (s) in EMD and Kusa cells are useful for this investigation. Porcine recombinant amelogenin stimulated the differentiation of Kusa cells indicating the possibility that amelogenin is the effective molecule.Although further studies are necessary, we could apply recombinant amelogenin to periodontal tissue regeneration.

本研究的目的是表征控制牙周韧带（PDL）细胞分化和功能的分子。构建了牛PDL组织cDNA文库，克隆了钙结合蛋白S100A4的cDNA。我们观察到S100A4 mRNA在突牙PDL中的表达水平较高，并且PDL细胞分泌该蛋白，该蛋白位于胶原纤维附近。在成骨细胞培养中加入重组S100A4蛋白抑制矿化。在培养中，机械应力作用下的PDL细胞增加了S100A4 mRNA和细胞骨架蛋白的表达。尽管该蛋白在MC3T3-E1细胞（成骨细胞）中的表达水平较低，但抑制MC3T3-E1细胞中S100A4的表达可刺激培养中的矿化。这些结果强烈表明S100A4是PDL组织中mermerization的抑制剂。另一方面，猪牙釉质基质衍生物（EMD）刺激牛PDL细胞和小鼠骨髓库萨细胞的生长和分化，从而刺激这些细胞的矿化。对成骨细胞的刺激作用也有助于牙周组织的再生。阐明EMD和库萨细胞中的有效分子对研究有重要意义。猪重组淀粉原蛋白刺激了库萨细胞的分化，表明淀粉原蛋白可能是有效分子。虽然还需要进一步的研究，但我们可以将重组淀粉原蛋白应用于牙周组织再生。

项目成果

Ikeda K,Kuroda S,Oida S,Iimura T,Duarte WR,Ohya K,Ishikawa I,Kasugai S: "rhFGF4 promotes ectopic bone formation induced by rhBMP2"Journal of Bone and Mineral Research. 15(suppl.1). S257 (2000)

Ikeda K、Kuroda S、Oida S、Iimura T、Duarte WR、Ohya K、Ishikawa I、Kasugai S：“rhFGF4 促进 rhBMP2 诱导的异位骨形成”《骨与矿物质研究杂志》。

春日井昇平: "Field遺伝子治療に向けた最新の研究から 骨髄ストローマ細胞の臨床応用"the Quintessence. 19・1. 210 (2000)

Shohei Kasugai：“来自现场基因治疗最新研究的骨髓基质细胞的临床应用”Quintessence 19・1（2000）。

Yoneda S,Kasugai S,Itoh D,Umezawa A,Kondo H,Kuroda S,Ohya K,Ohyama T: ".Effect of enamel matrix derivative (EMDOGAIN) on osteoblastic cells"Journal of Dental Research. 79(Special Issue). 474 (2000)

Yoneda S、Kasugai S、Itoh D、Umezawa A、Kondo H、Kuroda S、Ohya K、Ohyama T：“牙釉质基质衍生物 (EMDOGAIN) 对成骨细胞的影响”牙科研究杂志。

Duarte WR,Mikuni-Takagaki Y,Kawase T,Iimura T,Oida S,Ohya K,Takenaga K,Ishikawa I.Kasugai S: "Effects of mechnical stress on the mRNA expression of S100A4 and cytoskeletal components by periodontal ligament cells"Journal of Medical and Dental Sciences. 46

Duarte WR、Mikuni-Takagaki Y、Kawase T、Iimura T、Oida S、Ohya K、Takenaga K、Ishikawa I.Kasugai S：“机械应力对牙周膜细胞 S100A4 和细胞骨架成分 mRNA 表达的影响”杂志

Duarte WR,池田かおり,石川烈,春日井昇平,大谷啓一,飯村忠浩,大井田新一郎: "歯周組織におけるS100A4カルシウ結合タンパクの発現とその生理的役割"口腔病学会雑誌. 66. 223 (1999)

Duarte WR、Kaori Ikeda、Retsu Ishikawa、Shohei Kasugai、Keiichi Otani、Tadahiro Iimura、Shinichiro Oida：“牙周组织中 S100A4 钙结合蛋白的表达及其生理作用”口腔学会杂志 66. 223 (1999)。