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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.

本研究旨在揭示体内和体外破骨细胞分化过程中破骨细胞表面黏附分子的表达情况。将骨髓细胞与成骨细胞和骨髓细胞共培养，于培养第1、3、5天分离为贴壁细胞和非贴壁细胞，用流式细胞仪检测骨髓细胞表面抗原(Mac-1、LFA-1、CD44和MEL 14)，并对培养14天的骨髓细胞进行抗酒石酸酸性磷酸酶(TRAP)染色。为了解黏附分子(Mac-1、LFA-1、ICAM-1)在小鼠牙周组织中的表达情况，采用免疫组织化学方法对小鼠牙周组织进行了体内免疫组织化学染色。结果表明：体外实验中，破骨细胞黏附分子(Mac-1、LFA-1、ICAM-1)的表达弱于巨噬细胞。然后在体内实验中，在小鼠的牙周韧带内部和骨髓间隙中可以识别出Mac-1和LFA-1阳性细胞。ICAM-1阳性细胞主要分布在牙根尖部和髓腔周围。而正畸力作用于牙齿后，牙槽骨受压侧破骨细胞数量和黏附分子阳性细胞数均显著增加，提示黏附分子表达与破骨细胞分化之间的关系对破骨细胞向成骨细胞的成熟过程具有重要作用。