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破坏的牙周组织的再生，并且是功能的原始形式和恢复。人们认为，新骨，新的胶质和新结缔组织的重新粘附是组织被破坏的重生所必需的。我们先前报道说，球体是用牙周韧带成纤维细胞产生的，可用于牙周重建治疗。因为球体具有球形3维结构，并且包含具有细胞增殖活性并产生细胞外基质蛋白的细胞。这段时间，我们使用人肺泡骨衍生的细胞（HABC）与含有胎儿的胎儿小腿培养基（D-MEM）产生了球体（D-MEM）。我们使用针对胶原蛋白I型，骨桥蛋白（OP），骨钙蛋白（OC）和KI-67抗体进行了一项免疫组织化学研究，对球体进行了免疫组织化学研究。另一种方法是建立具有生长因子的培养方法，该方法是重组人血小板衍生的生长因子（RHPDGF）和无血清中的重组人B纤维细胞生长因子（RHB-FGF），在无血清中引入牙周组织再生治疗与斑点的正面反应。在第七天和21^<st>天。在拳头天观察到胶原蛋白I型在21^<ST>日观察到，并且在整个球体中观察到碱性磷酸酶的第一天到7^<th>天。 HABC可以在21^<ST>天在球体内表面产生OC和OP。含有100 ng/ml的Rhb-FGF的无血清培养基的培养速率最高的生长速率为42.0％的培养时间为5天的D-MEM。 HABC在球体环境中表示胶原蛋白I型，OC和OP，因此球体可能能够应用于牙周重建疗法。