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Study of bone-forming factors and their clinical evaluation

成骨因子的研究及其临床评价

基本信息

批准号：
04454496
负责人：
AMAGASA Teruo
金额：
$ 4.22万
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 General Scientific Research (B)
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454496/
关键词：
bone-forming factors organ culture insulin bone fracture healing rat calvaria clinicostatistical study

项目摘要

1.Serum-free bone-forming organ culture system using twenty-day fetal rat parietal bones was established. Bones were cultured on grids and placed on a rocking platform under the aerobic condition. This condition increased the bone-forming activities of cultured rat parietal bones. The calcium content, dry weight of bone, and lactate produced by bone tissue in the medium were measured to study the precise bone-forming process of vital bones.2.Using this improved bone-forming organ culture system, the effects of insulin on bone formation were studied. Insulin increased calcium content and dry weight of bone. These effects were maximum in bones treated with insulin at 10^<-6>M.Histomorphometrical analysis showed that the areas of mineralized bone and bone matrix increased, and that many osteoblasts and few osteoclasts appeared in bones treated with insulin at 10^<-6>Mor higher.Insulin inhibited the release of ^<45>Ca into the medium. From these results, it was suggested that insulin has a stimulatory effect on bone formation enhancing both bone mineralization and bone matrix formation.3.Cloning of bone morphogenetic protein (BMP) cDNA by PCR method was performed. Ectopic bone formation induced by BMP,was investigated, and also the molecular cloning of BMP type I receptor was done.4.In situ hybridization of the rat tibial bone marrow cvity, in which bone formation and resorption were indluced by colchicine, demonstrated that osteopontin mRNA was expressed during bone resorption process.5.Clinicostatistical and long-term follow-up studies of the pediatric fracture cases were performed.

1.建立了20d胎鼠顶骨无血清成骨器官培养体系。骨在网格上培养，并在有氧条件下放置在摇摆平台上。这种情况增加了培养的大鼠顶骨的骨形成活动。通过测定培养液中钙含量、骨干重、骨组织产生乳酸等指标，研究活骨的精确成骨过程。2.利用改进的成骨器官培养系统，研究胰岛素对成骨的影响。胰岛素增加骨钙含量和骨干重。组织<-6>形态计量学分析表明，在10 μ莫尔更高浓度胰岛素处理的骨中，矿化骨和骨基质面积增加，出现许多成骨细胞和少数破骨细胞<-6>，胰岛素抑制^ Ca释放<45>到培养基中。结果表明，胰岛素具有促进骨形成的作用，能促进骨矿化和骨基质的形成。3.用PCR方法克隆骨形态发生蛋白（BMP）cDNA。4.用秋水仙素诱导大鼠胫骨骨髓腔骨形成和骨吸收，原位杂交证实骨桥蛋白mRNA在骨吸收过程中表达。5.对儿童骨折病例进行临床统计学和长期随访研究。