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Research of the Mechanism of Calcific Deposition in Human.

人体钙化沉积机制的研究。

基本信息

批准号：
63440056
负责人：
HOSHINO Takashi
金额：
$ 5.38万
依托单位：
Dokkyo Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (A)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

项目摘要

[ Material and Method ]Two parallel gutters (interval : 25mm) were digged to the subchondral bone from the cartilage surface of tibial articular slice (depth : 30mm) of young cows (average 2.5 years old). One was filled with sodium pyrophosphate crystal and the other was filled with calcium chloride and keep the slice in room temparature for five days. White zone was formed between two gutters, 9mm apart from the gutter filled with sodium pyrophate. This white zone was analyzed by 1. Hematoxylin-Eosin stain, Von Kossa stain 2. SEM 3. Crystallographic studies by X-ray diffraction, infrated spectroscopy and Raman analysis 4. Enzymatic analysis by Kurata and Maeda method.[ Results ]1. The matrix from tangential to transitional layer was calcified. 2. Pavementlike crystals were detected by SEM. 3. Calcium salts were Ca_3(po_4)_2 or Ca_5(po_4)_3OH by crystallographic analyses. 4. Inorganic pyrophosphatase was detected in the matrix and cartilage cells below radial zone and in cartilage cells of superficial layer. Some experiments on the cartilage treated by sodium fluoride revealed neither white zone nor the evidence of inorganic pyrophosphatase activites.[ Points ]Calcium phosphate was formed in cartilage from sodium pyrophosphate and calcium chloride. The fact showed that articular cartilage acted as a gel media in which chemicals are able to diffuse and crystals can precipitate. Calcium pyrophosphate cyrstals may be precipitated in condition that inorganic pyrophosphatase is absent.

[材料与方法]取青年奶牛(平均2.5岁)的胫骨关节片软骨面，在软骨表面挖出两条平行沟(间距25 mm)至软骨下骨。一种填充焦磷酸钠晶体，另一种填充氯化钙，将切片在室温下保存5d。在两条沟之间形成白色区域，距离填有焦磷酸钠的沟9 mm。用苏木精-伊红染色、Von Kossa染色、扫描电子显微镜、X-射线衍射仪、红外光谱和拉曼光谱对白带进行了分析，用仓田法和前田法进行了酶促分析。2.扫描电子显微镜观察到铺片状晶体。3.结晶分析表明，钙盐为Ca_3(PO_4)_2或Ca_5(PO_4)_3OH。4.无机焦磷酸酶存在于放射带下方的基质和软骨细胞以及浅层软骨细胞中。用氟化钠处理的软骨，既没有白带，也没有无机焦磷酸酶活性的证据。软骨中有焦磷酸钠和氯化钙生成磷酸钙。事实表明，关节软骨作为一种凝胶介质，化学物质能够在其中扩散，晶体可以沉淀。在没有无机焦磷酸酶的条件下，可以析出焦磷酸钙晶体。