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Basic Research for Tissue Engineering of Periodontal Tissue Regeneration : Validity of Enamel Matrix Proteins and Periodontal Ligament Fibroblast Synthesized Proteins.

牙周组织再生的组织工程基础研究：牙釉质基质蛋白和牙周膜成纤维细胞合成蛋白的有效性。

基本信息

批准号：
14370714
负责人：
KAWASE Toshio
金额：
$ 8.51万
依托单位：
Kanagawa Dental College
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

Enamel matrix proteins (enamel matrix derivative, Emdogain^<【○!R】>, EMD) have been clinically used regenerate the periodontal tissues. Although it is clamed that EMD can be used to promote new attachment formation around periodontally invoved teeth, the underlying biological mechanism is not understood. We have demonstrated that the secreted proteins of HPLF have a potency for the cell attachment and spreading activity factor (CASF) promoting tissue regeneration.The present study investigated the effect of EMD on the behavior of rat bone marrow cells(RBMC) in vitro and compared it with that for the conditioned medium protein(CMP) obtained from human periodontal ligament-derived fibroblasts (HPLF), especially their attachment property, alkaline phosphatase(ALPase) activity and the mRNA levels for ALPase, bone sialoprotein(BSP), Osteopontin(OPN), and osteocalcin(OCN). The hydrophobic dishes were coated EMD (25μg/ml) or CMP (25μg/ml) for 24 hrs at 4℃. The dishes were treated with heat inactivated BSA.RBMCs were obtained from the femora of 7-week-old male rats and cultured in α-MEM containing 10% FCS and 0.25mM ascorbic acid phosphate on hydrophobic culture plates coated with EMD or CMP. RBMCs attached and spread within 3 hours. RBMCs cultured onto EMD coated dishes expressed high ALPase activity and mRNA levels when compared with CMP coated ones. Furthermore, the expression of OPN and OCN mRNAs of RBMCs was significantly enhanced by the presence of EMD. EMD did not play a chemotactic activity. CMP stimulate the migration activity of HPLF.Our results indicated that RBMCs responded differently to EMD or CMP, and that EMD stimulated osteogenic differentiation of RBMCs. Therefore, HPLF may interacted with EMD coated on dentin surface, proliferate and differentiate. And mesenchymal cells (RBMC) couled be migrated by CMP and differentiated to osteogenic cells.

釉质基质蛋白（釉质基质衍生物，Emdogain^[0！R]>、EMD）在牙周组织再生中的应用。尽管有研究认为EMD可以促进牙周损伤牙周围新附着体的形成，但其生物学机制尚不清楚。本研究观察了EMD对体外培养的大鼠骨髓细胞（RBMC）行为的影响，并与人牙周膜成纤维细胞（HPLF）的条件培养基蛋白（CMP）进行了比较，特别是对细胞粘附和铺展活性因子（CASF）促进组织再生的作用。碱性磷酸酶（ALP 3）活性和ALP 3、骨唾液蛋白（BSP）、骨桥蛋白（OPN）和骨钙素（OCN）的mRNA水平。疏水皿用EMD（25μg/ml）或CMP（25μg/ml）在4℃下包被24小时。取7周龄雄性大鼠股骨，用含10%FCS和0.25mM抗坏血酸磷酸盐的α-MEM培养液培养于EMD或CMP包被的疏水培养板上。RBC在3小时内附着并扩散。与CMP包被的相比，EMD包被的培养皿上培养的RBMCs表达高的ALP 3活性和mRNA水平。此外，EMD的存在显著增强了RBMCs的OPN和OCN mRNA的表达。EMD不具有趋化活性。结果表明，EMD和CMP对红细胞的迁移有不同的影响，EMD能促进红细胞的成骨分化。因此，HPLF可能与包被在牙本质表面的EMD相互作用，增殖和分化。骨髓间充质细胞（RBMC）可被CMP迁移并向成骨细胞分化。