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Investigation and application of new recombinant materials inducing periodontal tissue regeneration

诱导牙周组织再生的新型重组材料的研究及应用

基本信息

批准号：
18592273
负责人：
TANAKA Akio
金额：
$ 2.44万
依托单位：
Osaka Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18592273/
关键词：
Dentistry Regenerative medicine Pathology Cell Tisue Biomolecule 細胞組織

项目摘要

Research conducted in 2007 revealed that the optimal conditions for peptide to form hard tissues were 7.5-15mg/mL for peptide concentration and 7-14 days for time interval after peptide injection into the back skins of rats. In the tissue reaction to new付ype Emdogain[○!R], eosinophilic amorphous substances (EAS) and many mononuclear cells were observed about 7 days after injection of peptide into the rat backs, and on day 14 a small amount of EAS remained, mononuclear cells and fibroblasts were increased in number, and scattered eosinophils were observed. However, hard tissue formation was not confirmed. Ultrastructurally, some cells showed abundant rough endoplasmic reticulum, and the others many mitochondria. In studies of the effect of peptide on human periodontal ligament fibroblasts (HPdLF), alkaline phosphatase activity after 7 days of culture was increased 8-fold as compared with that after one day of culture. Total RNA was extracted 1 and 3 days after culture of HPdLF with 100mg/mL of peptide, and cDNA reverse transcription was carried out, followed by microarray analysis. On culture day 3, compared with on day 1, mRNA of bone morphogeneticprotein receptor type 1(BMPR1) Awas increased 34.18-fold, that of BAP41.97-fold, that of osteonectin 1.55-fold, and that of BMPR 1B 1.05-fold, but that of fibroblast growth factor receptor-like protein 1(FGFRL1) was reduced to 1/25. Thus, hard tissue formation was observed in the tissue reaction to peptide, BMPR 1A gene expression, which is associated with bone formation, was increased to a high level, and that of FGFRL1 was markedly reduced. These results might indicate that the peptide make HPdLF induce the hard tissue formation.

2007年的研究表明，多肽形成硬组织的最佳条件是多肽浓度为7.5-15 mg/mL，多肽注射到大鼠背部皮肤后间隔7-14天。在对新付[0！R]的组织反应中，大鼠背部注射多肽后7d左右可见嗜酸性无定形物质(EAS)和大量单核细胞，第14天仍有少量EAS残留，单核细胞和成纤维细胞增多，可见散在的嗜酸性粒细胞。然而，硬组织的形成还没有得到证实。超微结构上，有的细胞粗面内质网丰富，有的细胞线粒体较多。在多肽对人牙周膜成纤维细胞(HPdLF)影响的研究中，培养7d的碱性磷酸酶活性是培养1d的8倍。含100 mg/mL肽的HPdLF培养1天和3天后提取总RNA，进行cDNA逆转录后进行基因芯片分析。培养3d时，骨形态发生蛋白受体1型(BMPR1)A、BAP41.97、On-ectin、BMPR-1B的mRNA水平分别为第1天的34.18倍、41.97倍、1.55倍、1.05倍，而成纤维细胞生长因子受体样蛋白1(FGFRL1)的水平仅为1/25。因此，在多肽的组织反应中观察到了硬组织的形成，与骨形成相关的BMPR1a基因的表达增加到高水平，而FGFRL1的表达明显减少。这些结果可能表明，HPdLF能诱导硬组织的形成。