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The molecular analysis of laser irradiation effect for the osteoblast

激光照射对成骨细胞影响的分子分析

基本信息

批准号：
13672176
负责人：
SASAGURI Kenichi
金额：
$ 2.3万
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：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672176/
关键词：
Osteoblast cbfal 骨芽細胞 cbfa 1 レーザー照射 Cbfa1

项目摘要

The study was investigated that what kind of effect the laser irradiation causes the human osteoblasts cell line, NHOst cell (Bio Whittaker Co.), in vitro. As a reason using the NHOst cell, it is for examining for the purpose of always feeding back the result for the daily clinic, such as regeneration of oral hard tissues and implantation. However, this cell showed morphology of dedifferentiating by the subculture of 2〜3 times, and alkaline phosphatase activity which was an index to the differentiation of osteoblast and DNA content which are an index to cell proliferation did not become constant. Then, the experiment should be carried out using MG63 cell that it is the human osteosarcoma cell line. It is reported that MG63 cell is efficiently differentiated an osteoblast like cell by the addition of active vitamin D_3, and we have been confirmed that this applicant has also expressed osteocalcin gene by the polymerase chain reaction (PCR). In the future, it will be examined that the effects of laser irradiation investigate on the osteoblast like cell, MG63.In the meantime, We have raised monoclonal antibody against recombinant murine cbfa1 and cbfβ protein. The specificity of these antibodies were characterized by its reaction in Western blot analysis and electrophoretic mobility shift assay (EMSA) on mourine cells and tissues. In the future, the specificity of the antibody is examined using MG63 cell and human hard tissues.

研究了激光照射对人成骨细胞系NHOst细胞（Bio Whittaker Co.）体外作为使用NHOst细胞的原因，其用于检查，目的是始终将结果反馈给日常临床，例如口腔硬组织的再生和植入。然而，该细胞通过2 ~ 3次传代培养显示出去分化的形态，并且作为成骨细胞分化指标的碱性磷酸酶活性和作为细胞增殖指标的DNA含量没有变得恒定。那么，实验应该使用MG 63细胞进行，它是人类骨肉瘤细胞系。据报道，MG 63细胞通过添加活性维生素D_3而有效地分化为成骨样细胞，并且我们已经通过聚合酶链反应（PCR）证实本申请人也表达了骨钙素基因。本实验同时制备了抗重组鼠cbf α 1和cbfβ蛋白的单克隆抗体。用免疫印迹法和电泳迁移率变动分析法（EMSA）对抗体的特异性进行了鉴定。在未来，使用MG 63细胞和人硬组织检查抗体的特异性。