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Fundamental research of application of self periodontium derived cells spheroid to guided tissue regeneration

自体牙周源细胞球体应用于引导组织再生的基础研究

基本信息

批准号：
14571991
负责人：
DEGUCHI Shinji
金额：
$ 1.86万
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 (C)
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

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571991/
关键词：
periodontum derived cells regeneration spheroid cell growth

项目摘要

The ultimate goal of periodontal therapy is the regeneration of destroyed periodontal tissues, and is in the original form and recovery of a function. It is thought that the re-adhesion by the new bone, new cementum, and new connective tissue is required for the destroyed rebirth of an organization. We previously reported that the spheroid was produced with periodontal ligament fibroblasts and might be used for periodontal reconstructive therapy. Because spheroid has spherical 3-dimensional structure and that contained the cells which have cell proliferation activity, and produced extracellular matrix proteins.This time, we produced spheroid using the human alveolar bone derived cell(HABC) with containing fetal calf serum medium(D-MEM). We performed an immunohistochemical study for spheroids using the monoclonal antibody against collagen type-I, osteopontin(OP), osteocalcin(OC) and Ki-67 antibodies. Another approach is to establish of the culture method with growth factor which were a recombinant human platelet derived growth factor(rhPDGF) and a recombinant human b-fibroblast growth factor(rhb-FGF), in the serum free medium to introducing the periodontal tissue regeneration therapy with the spheroids.The positive reaction of Ki-67 was observed in the whole of spheroid on the first day and the third day, but observed only in the surface layer on the seventh day and the 21^<st> day. Collagen type-I was observed on the fist day to on the 21^<st> day and an alkaline phosphatase was observed in the whole spheroid on the first day to 7^<th> day. HABC could produce OC and OP at inside surface of the spheroid on the 21^<st> day. The culture with serum free medium contained with 100 ng/ml rhb-FGF appeared the highest growth rate was 42.0 % of the culture with D-MEM at the 5 day. HABC expressed Collagen type-I, OC and OP in the spheroid environment, so spheroids may be capable of being applied to periodontal reconstructive therapy.

牙周治疗的最终目标是使受损的牙周组织再生，并恢复原有的形态和功能。人们认为，新骨、新牙骨质和新结缔组织的重新粘附是组织被破坏的再生所必需的。我们之前报道过该球体是由牙周膜成纤维细胞产生的，可能用于牙周重建治疗。由于球体具有球形3维结构，并且含有具有细胞增殖活性的细胞，并产生细胞外基质蛋白。这次，我们使用人牙槽骨来源细胞（HABC）和含有胎牛血清培养基（D-MEM）的球体制作。我们使用抗 I 型胶原、骨桥蛋白 (OP)、骨钙素 (OC) 和 Ki-67 抗体的单克隆抗体对球体进行了免疫组织化学研究。另一种方法是建立生长因子（重组人血小板衍生生长因子（rhPDGF）和重组人b-成纤维细胞生长因子（rhb-FGF）在无血清培养基中的培养方法，以球体引入牙周组织再生治疗。第一天和第三天在整个球体中观察到Ki-67阳性反应，但仅在表层观察到Ki-67阳性反应第七天和21^<st>天。在第一天到第21天观察到I型胶原，并在第一天到第7天在整个球体中观察到碱性磷酸酶。 HABC在第21^<st>天可以在球体内表面产生OC和OP。在第5天时，用含有100ng/ml rhb-FGF的无血清培养基进行的培养出现了最高的生长率，为用D-MEM进行的培养的42.0％。 HABC在球体环境中表达I型胶原、OC和OP，因此球体可能能够应用于牙周重建治疗。