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identification of Growth and Differentiation Factors in Enamel Matrices andApplication of These Factors to Periodontal Tissue Regeneration

牙釉质基质中生长和分化因子的鉴定及其在牙周组织再生中的应用

基本信息

批准号：
13470460
负责人：
KASUGAKI Shohei
金额：
$ 9.09万
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：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

项目摘要

The purpose of the present study was to identify growth and differentiation factors in enamel matrices and to apply these factors to periodontal tissue regeneration.EMDOGAIN, a commercially available product, which is derived from porcine enamel matrices, stimulated proliferation and osteoblastic differentiation of KUSA cells (mouse bone marrow derived cells). Furthermore, it promoted bone regeneration in rats. Results in the experiments with the antibodies against BMP-2 and TGF-b indicated that EMDOGAIN contains these factors; however, BMP activity of EMDOGAIN is not remarkable because subcutaneous implantation of EMDOGAIN did not induce ectopic bone formation. It is possible that an effective factor in EMDOGAIN is amelogenin; however, amelogenin did not promote bone regeneration in rats. Therefore, it is likely that EMDOGAIN contains various effective factors and the combination of these factors promotes periodontal tissue regeneration.Biochemical separation of effective factors from porcine enamel matrices has not been accomplished because of low solubility of porcine enamel matrices. Thus, we developed unique gene transfer method with plasmid expression vector. Furthermore, we extracted RNA from mouse incisol tooth germs and subcutaneous tissue and compared gene expression with DNA chips. Genes, which are specifically expressed in cementoblasts, would be useful markers of cementoblast differentiation. To identify cementoblast specific gene, we dissected cementoblasts and periodontal ligament cells from cryo-sections with laser-capture microscope and extracted RNA. Although the amount of the available RNA was very small, we confirmed the possibility of the analysis with DNA chips after the amplification. Promoting these studies would proceed to the effective way of periodontal tissue regeneration in the near future.

本研究的目的是鉴定釉质基质中的生长和分化因子，并将这些因子应用于牙周组织再生。EMDOGAIN是一种市售产品，来源于猪釉质基质，刺激KUSA细胞（小鼠骨髓来源的细胞）的增殖和成骨细胞分化。此外，它还促进了大鼠的骨再生。用抗BMP-2和TGF-β的抗体进行的实验结果表明，EMDOGAIN含有这些因子;然而，EMDOGAIN的BMP活性并不显著，因为皮下植入EMDOGAIN不会诱导异位骨形成。EMDOGAIN中的一个有效因子可能是釉原蛋白;然而，釉原蛋白并不促进大鼠的骨再生。因此，很可能EMDOGAIN含有各种有效因子，这些因子的组合促进牙周组织再生。由于猪牙釉质基质的溶解度低，尚未完成从猪牙釉质基质中分离有效因子的生化分离。因此，我们开发了独特的基因转移方法与质粒表达载体。此外，我们从小鼠门牙牙胚和皮下组织中提取RNA，并与DNA芯片比较基因表达。在成牙骨质细胞中特异表达的基因将成为成牙骨质细胞分化的有用标记。为了鉴定成牙骨质细胞特异性基因，我们用激光捕获显微镜从冰冻切片中分离成牙骨质细胞和牙周膜细胞，提取RNA。尽管可用RNA的量很少，但我们证实了扩增后用DNA芯片分析的可能性。促进这些研究将在不久的将来为牙周组织再生的有效途径进行研究。

项目成果

期刊论文数量（72）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Yokozeki M, Afanador E, Nishi M, Kaneko K, Shimokawa H, Yokote K, Deng C, Tsuchida K, Sugino H, Moriyama K: "Smad3 is required for enamel biomineralization."Biochem Biophys Res Commun.. 305(3). 684-690 (2003)

Yokozeki M、Afanador E、Nishi M、Kaneko K、Shimokawa H、Yokote K、Deng C、Tsuchida K、Sugino H、Moriyama K：“牙釉质生物矿化需要 Smad3。”Biochem Biophys Res Commun. 305(3)。