The Adhesion molecule expression in osteoclastic differentiating process

破骨细胞分化过程中粘附分子的表达

基本信息

批准号：
03670969
负责人：
MITANI Hideo
金额：
$ 1.28万
依托单位：
TOHOKU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1992
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03670969/
关键词：
OSTEOCLAST MAC-1 LFA-1 CD 44 MEL 14 ICAM-1 FACS IMMUNOHISTOCHEMISTRY Mel 14 骨髄脾臓末梢血破骨細胞分化因子 Macー1抗原 LFAー1抗原混培養上清

项目摘要

The purpose of this investigation was to reveal the expressions of adhesion molecules on the surface of osteoclast during osteoclastic differentiation in vivo and in vitro. Bone marrow cells were cultured with the co-culture fluid of osteoblasts and bone marrow cells and divided to adherent cells and non adherent cells at 1,3,5 days.Surface antigens (Mac-1,LFA-1,CD 44 and Mel 14) of the cultured bone marrow cells were analyzed by Flow cytometry (FACS) and also the cultured bone marrow cells were stained with tartrate resistent acid phosphatase (TRAP) on 14 days culture. In order to evaluate the expressions of adhesion molecules (Mac-1,LFA-1,ICAM-1) in peridontal tissues of mice, immunohistochemical staining in vivo were made.The following results were obtained:In vitro experiments,adhesion molecules (Mac-1,LFA-1,ICAM-1) of osteoclasts converted to weaker expression than that of macrophages in bone marrow cells cultured with the coculture fluid. And then in vivo experiments, Mac-1 and LFA-1 positive cells could be recognizes in the inside of periodontal ligaments and the marrow spaces of mice. ICAM-1 positive cells were observed specifically around the apex of the tooth root and marrow spaces. However,when orthodontic force were applied to teeth, in compressed side of alveolar bone, both numbers of osteoclast and the adhesion molecule positive cells were significantly increased.These results suggested that the relationship between the expressions of adhesion molecules and the differentiation of osteoclasts was very important for the maturation process of osteoclast from hematopoietic stem cells.

本研究的目的是揭示体内和体外破骨细胞分化过程中破骨细胞表面粘附分子的表达。骨髓细胞与成骨细胞和骨髓细胞的共培养液一起培养，并在第1、3、5天分为贴壁细胞和非贴壁细胞。表面抗原（Mac-1、LFA-1、CD 44和Mel 14）通过流式细胞术（FACS）分析培养的骨髓细胞，并且还用抗酒石酸酸性磷酸酶（TRAP）对培养的骨髓细胞进行14染色天文化。为了评价小鼠牙周组织中粘附分子(Mac-1、LFA-1、ICAM-1)的表达情况，进行了体内免疫组化染色，得到如下结果：体外实验中，粘附分子(Mac-1、LFA-1、ICAM-1)在小鼠牙周组织中的表达情况。在用共培养液培养的骨髓细胞中，破骨细胞的1、LFA-1、ICAM-1)的表达比巨噬细胞的表达弱。然后在体内实验中，在小鼠牙周膜内部和骨髓腔中可以识别出Mac-1和LFA-1阳性细胞。在牙根尖和骨髓腔周围特别观察到 ICAM-1 阳性细胞。然而，当对牙齿施加正畸力时，在牙槽骨受压侧，破骨细胞和粘附分子阳性细胞的数量均显着增加。这些结果表明，粘附分子的表达与破骨细胞的分化之间存在密切关系。对于造血干细胞破骨细胞的成熟过程很重要。