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Ultrastructural and histochemical studies on the coupling phenomena between bone formation and bone resorption

骨形成与骨吸收耦合现象的超微结构和组织化学研究

基本信息

批准号：
62480370
负责人：
OZAWA Hidehiro
金额：
$ 4.42万
依托单位：
Niigata 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：
1987
资助国家：
日本
起止时间：
1987 至 1989
项目状态：
已结题

项目摘要

The basic objection of this study is to elucidate the fine structural and cytochemical characteristics of the coupling phenomena between bone resorption and formation. The first objective was designed to elucidate The cyto chemical events between the osteoclastic bone resorption and subsequent osteoblastic bone formation. The ACPase positive resorbed bone surface continued to the cement line, on which osteoblasts were induced and formed a new bone. This suggests that the osyteoclasts may move as a resorbing bone and form a cement line, which may be a responsible for the coupling of the bone resorption and formation. The second objective was to reveal fine structural and cytochemical details on relationships between osteoclastic and osteoblastic cells in the differentiation and activation processes of osteoclasts. At first, LM and EM autoradiography(A.R) was performed on an organ culture using [^<125>I] calcitonin(CT) to identify osteoclastic cells.Developed silver grains of [^<125>I]-C … More T were located on the TRACP-positive mono and multinucleated cells. In the EM-A.R, silver grains could be observed on the Golgi bodies and plasma membranes of both osteoblast progenitors and osteoclasts, suggesting that[^<125>I]-CT binds to receptors of plasma memoranes of osteoclastic cells, then internalized and turned to reach to the Goligi bodies. Ultrastructural and lectin-cytochemical observations were then carried out. Osteoblastic cells had close contact with differentiating osteoclasts, where focal connections and coated pits could frequently be observed in both osteoclastic and osteoblastic cells facing each other.When mouse spleen cells were co-cultured with freshly isolated osteoblasts in the presence of 1,25-(OH)_2D_3, TRACF-positive multinuclear cells were formed. In this case, osteoblasts had also close contact with spleen cells in the same way as shown in vivo. These results indicate that osteoblastic cells are required for the differentiation of osteoclast orogenitors into osteoclasts by way of direct cell-cell contact, and this intimate cell contact presumably enables the osteoclast progenitors to respond to stimuli present on the osteoblastic cell surface. Lectin reaction was seen on Plasma membranes both in developing osteoclastic and adjacent osteoblastic cells suggesting that sugar residues on the plasma membrane may play a role in the cellular recognition via receptors, and also may be involved in concentrating appropriate amount of several local factors. Less

本研究的基本目的是阐明骨吸收和骨形成之间耦合现象的精细结构和细胞化学特征。第一个目的是阐明成骨细胞骨吸收和随后的成骨细胞骨形成之间的细胞化学事件。ACEI阳性骨吸收表面延续至骨水泥线，在骨水泥线上诱导成骨细胞形成新骨。这表明破骨细胞可能作为吸收骨移动并形成骨水泥线，这可能是骨吸收和骨形成耦合的原因。第二个目标是揭示在破骨细胞的分化和活化过程中骨细胞和成骨细胞之间的关系的精细结构和细胞化学细节。首先，使用[^ I]降钙素（CT）对器官培养物进行LM和EM放射自显影（A.R）<125>以鉴定骨细胞。<125> ...更多信息 T定位于TRACP阳性单核和多核细胞上。在EM-A.R中，在成骨祖细胞和破骨细胞的高尔基体和质膜上都可以观察到银颗粒，这表明[^<125>I]-CT与成骨细胞的血浆记忆受体结合，然后内化并到达Goligi体。然后进行超微结构和凝集素细胞化学观察。成骨细胞与分化中的破骨细胞接触密切，成骨细胞与成骨细胞相互面对时，常可观察到灶状连接和被膜小凹，小鼠脾细胞与新鲜分离的成骨细胞在1，25-（OH）_2D_3存在下共培养时，可形成TRACF阳性的多核细胞。在这种情况下，成骨细胞也以与体内所示相同的方式与脾细胞紧密接触。这些结果表明，破骨细胞orogenitors分化成破骨细胞所需的直接细胞-细胞接触的方式，这种亲密的细胞接触，大概使破骨细胞祖细胞对刺激的成骨细胞表面上。在发育中的成骨细胞和邻近的成骨细胞的质膜上均观察到凝集素反应，这表明质膜上的糖残基可能通过受体在细胞识别中起作用，并且还可能参与浓缩适量的几种局部因子。少