Livne E, Oliver C, Leapman R D, Rosenberg L C, Poole A R, Silbermann M
Musculoskeletal Research Laboratory, Rappaport Family Institute for Research in the Medical Sciences, Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel.
J Anat. 1987 Feb;150:61-74.
A combined approach of light microscopy, immunofluorescence, transmission electron microscopy and electron energy loss spectroscopy (EELS) was used to study age-related changes in the condylar cartilage in mice. Chondrocalcin, a cartilage matrix calcium-binding protein, was demonstrated by indirect immunofluorescence microscopy using monospecific antibodies. In one week old animals the most intense staining was observed in the matrix around the hypertrophic cells in the mineralising zone, to a lesser degree around the cells in the zone of chondroblasts, while no staining was noted in the zone of chondroprogenitor cells and in the matrix around the early hypertrophic cells. In the mineralisation zone the distribution of chondrocalcin correlated with that of mineral deposits as revealed by the von Kossa stain. The matrix between the early hypertrophic cells as shown by transmission electron microscopy revealed the presence of matrix vesicles and demonstrated a gradual accumulation of hydroxyapatite in the mineralising zone. In one month old animals chondrocalcin localisation was mainly confined to the lower hypertrophic zone which also demonstrated positive von Kossa staining was seen along the articular surface. In older animals multiple electron-dense structure that resembled matrix vesicles were observed in the non-mineralising portions of the condylar cartilage. Use of the EELS method confirmed the almost complete lack of calcium ions in these structures. In contrast, with the use of the same method, detectable amounts of calcium were recorded in vesicles in the mineralising zones of all age groups. Hence what appear ultrastructurally as structures similar to matrix vesicles represent atypical vesicles that might characterise an ageing and degenerative articular cartilage and are not necessarily associated with the mineralisation process.
采用光学显微镜、免疫荧光、透射电子显微镜和电子能量损失谱(EELS)相结合的方法,研究小鼠髁突软骨的年龄相关变化。软骨钙素是一种软骨基质钙结合蛋白,使用单特异性抗体通过间接免疫荧光显微镜进行检测。在1周龄的动物中,在矿化区肥大细胞周围的基质中观察到最强的染色,在成软骨细胞区细胞周围的染色程度较轻,而在软骨祖细胞区和早期肥大细胞周围的基质中未观察到染色。在矿化区,软骨钙素的分布与用冯·科萨染色显示的矿物质沉积分布相关。透射电子显微镜显示,早期肥大细胞之间的基质中存在基质小泡,并表明矿化区中羟基磷灰石逐渐积累。在1月龄的动物中,软骨钙素的定位主要局限于较低的肥大区,该区域在关节表面也显示出阳性冯·科萨染色。在老年动物中,在髁突软骨的非矿化部分观察到多个类似基质小泡的电子致密结构。使用EELS方法证实这些结构中几乎完全缺乏钙离子。相反,使用相同的方法,在所有年龄组的矿化区小泡中记录到可检测量的钙。因此,超微结构上看似与基质小泡相似的结构代表非典型小泡,可能是老化和退变关节软骨的特征,不一定与矿化过程相关。