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A Cytobiochemical Characterization of Cell Diferentiation Derived from Calcified and Non-calcified Tissues.

钙化和非钙化组织细胞分化的细胞生化表征。

基本信息

批准号：
59440078
负责人：
SAITO Shigeru
金额：
$ 12.48万
依托单位：
Kanagawa Dental College
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (A)
财政年份：
1984
资助国家：
日本
起止时间：
1984 至 1986
项目状态：
已结题

项目摘要

The mechanism of calcification was investigated in terms of characteristic cell function of periodontal ligament cell (PLF) and fibroblast derived from non-calcified tissues in comparison with osteogenic cells from rat pup calvaria and bone in which extensive researches had been accumulated during past decays.The periodontal ligament is mainly of fibrous tissue containing various non-collagenous components, which tightly connects the tooth to the alveolar bone. Although the oxygen uptake of periodontal ligament was measured as 1/50 to that liver ( <Q_O2> /DNA=0.2), the metabolic activities of amino acids, proteins and glycosaminoglycans synthesis were found extensive through transamination and gluconeogenesis of intermediates converted from succinate-1,4- <^(14)C> which was added in the medium of short-term incubation system in vitro. Bovine PLF has a remarkably high activity of alkaline phosphatase (ALPase) compared to skin fibroblasts, which seem to have similar, but not identical phenotype with that of the rat osteoblast-like cells. The ALPase prepared from the human PLF and alveolar bone exhibited the same enzymatic character with bovine PLF. It is now possible to refer to PLF as an osteoblastic fibroblast and has proposed this as a work hypothesis. Important findings of this research are that ALPase activity of PLF and bone cells was regulated by 1,25 <(OH)_2> <D_3> and dexamethasone, and also 66Kd phosphoprotein was found as dose responding protein of active <D_3> within physiological concentration, which may have an important role for the transformation of octacalcium phosphate. Furtheremore, three stains of monoclonal antibodies synthesized cell line for human PLF were obtained by the cell fusion using myenoma NS-S.Those results mentioned above provide new aspect for the processes of calcification which mainly consists of inhibitory mechanism of mineralization observed in periodontal ligament.

从非钙化组织来源的牙周膜细胞（PLF）和成纤维细胞的特征性细胞功能方面探讨了钙化的机制，并与近年来研究较多的大鼠颅骨成骨细胞和骨成骨细胞进行了比较。牙周膜主要是纤维组织，含有各种非胶原成分，将牙齿与牙槽骨紧密连接。虽然牙周膜的氧摄取量仅为肝脏的1/50（<Q_O2>/DNA=0.2），但在体外短期培养体系中，加入琥珀酸-1，4- <^（14）C>转化的中间产物，通过转氨作用和代谢产物的生成，发现牙周膜具有广泛的氨基酸、蛋白质和糖胺聚糖合成代谢活性。与皮肤成纤维细胞相比，牛PLF具有显著高的碱性磷酸酶（ALP 3）活性，皮肤成纤维细胞似乎与大鼠成骨细胞样细胞具有相似但不相同的表型。由人PLF和牙槽骨制备的ALPase与牛PLF具有相同的酶特征。现在可以将PLF称为成骨细胞成纤维细胞，并将其作为工作假设。本研究的重要发现是：1，25 <（OH）_2>和地塞米松对PLF和骨细胞的ALP活性有调节<D_3>作用，同时发现66 Kd磷蛋白是生理浓度范围内活性的剂量反应蛋白<D_3>，可能对磷酸八钙的转化有重要作用。此外，还利用骨髓瘤NS-S细胞融合技术，获得了3株人PLF单克隆抗体合成细胞系。上述结果为牙周膜钙化过程中矿化抑制机制的研究提供了新的思路。