Quantitative determination of morphological and territorial structures of articular cartilage from both perpendicular and parallel sections by polarized light microscopy.
Quantitative determination of morphological and territorial structures of articular cartilage from both perpendicular and parallel sections by polarized light microscopy.
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DOI：
10.3109/03008207.2011.595521
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发表时间：
2011
影响因子：
2.9
通讯作者：
Bidthanapally A
中科院分区：
文献类型：
--
作者：
Mittelstaedt D;Xia Y;Shmelyov A;Casciani N;Bidthanapally A
为了研究关节软骨中胶原纤维的 3D 结构，将全层犬肱骨软骨切片成垂直切片，包括关节表面和软骨下骨，以及大约 100 个连续的平行切片，每个切片厚度为 6 μm，软骨深度不同。每个切片均使用具有 5 倍物镜（2.0μm 像素大小）的偏光显微镜 (PLM) 进行成像，生成两个定量图像（角度和延迟）。还使用 40 倍物镜（0.25μm 像素大小）对选定的切片进行成像。在距关节表面的深度增加时，垂直截面中的角度和延迟结果显示出表面和深层软骨之间原纤维方向众所周知的 90° 变化。相比之下，平行部分的延迟结果从关节表面开始减小，并且在大部分径向区域中保持近似于零，而平行部分的角度结果仅改变约30°。 61 个软骨细胞簇周围的区域基质形态通过其长度、纵横比和方向进行量化。表面软骨中的细胞簇在平行和垂直截面上均呈椭圆形。在径向区域中，细胞簇在垂直截面中定向为垂直柱，并在平行截面中定向为圆形分组。这种正交成像技术可以更好地了解关节软骨中的 3D 区域和区域间原纤维，其紊乱可能意味着骨关节炎等退行性软骨疾病的发生。
In order to investigate the 3D structure of the collagen fibrils in articular cartilage, full thickness canine humeral cartilage was microtomed into perpendicular sections that included both the articular surface and the subchondral bone and approximately 100 successive parallel sections that were each 6 μm thick and from a different cartilage depth. Each section was imaged using polarized light microscopy (PLM) with a 5x objective (2.0μm pixel size), generating two quantitative images (angle and retardation). Selected sections were also imaged using a 40x objective (0.25μm pixel size). At an increased depth from the articular surface, the angle and retardation results in the perpendicular sections showed the well-known 90° change in fibril orientation between the surface and deep cartilage. In contrast, the retardation results of the parallel sections decreased from the articular surface and remained approximately zero through most of the radial zone, while the angle results of the parallel sections only changed about 30°. The territorial matrix morphology surrounding 61 chondrocyte clusters was quantified by its length, aspect ratio, and orientation. The cellular clusters in the surface cartilage were ellipsoidal in both the parallel and perpendicular sections. In the radial zone, the cellular clusters were oriented in vertical columns in the perpendicular sections and as circular groupings in the parallel sections. This orthogonal imaging technique could provide a better understanding of the 3D territorial and interterritorial fibrils in articular cartilage, the disturbance of which could signify the onset of degenerative cartilage diseases such as osteoarthritis.