大鼠膝关节软骨不同染色方法的差异

背景：国内外学者对关节软骨对软骨进行了大量研究，需要用不同的染色方法对研究结果进行分析，但将多种染色方法同时应用于软骨研究及探讨染色机制的较少。目的：探讨不同染色方法对大鼠膝关节软骨染色的优缺点。方法：取正常大鼠膝关节软骨，行苏木精-伊红、番红O、阿尔辛蓝、甲苯胺蓝、番红-阿尔辛蓝、番红-固绿染色，观察软骨结构。结果与结论：苏木精-伊红、番红O、甲苯胺蓝染色均可观察到潮线，分别为蓝色、红色和蓝色，番红O和甲苯胺蓝染色强度朝潮线方向增强，番红O染色对潮线的观察优于其他染色方法。苏木精-伊红染色关节软骨4层结构清晰，软骨细胞呈柱状排列，基质呈均匀呈嗜碱性染色。番红O染色显示4层结构层次清楚，基质深层染色最红。阿尔辛蓝染色显示，pH 1.0时软骨细胞周边部分被阿尔辛蓝强烈染色，pH 2.5时阿尔辛蓝染色较深。甲苯胺蓝染色显示组织结构层欠清，胞核染色清晰，胞浆几乎不着色，基质呈淡蓝紫色。番红-阿尔辛蓝染色显示软骨表面及基质呈不均一的红色，深层颜色较深，软骨细胞周围蓝染；番红-固绿染色显示软骨基质呈均匀的红色，软骨下骨呈绿色，软骨组织与骨组织分对比鲜明。提示上述各种方法均可观察到软骨的四层结构，但以番红O染色显示软骨各层和潮线结构最佳；苏木精-伊红染色观察软骨细胞形态变化较其他方法清楚。

Different staining methods for rat knee articular cartilage

BACKGROUND: Extensive research has been made concerning articular cartilage by domestic and foreign scholars. Different staining methods are needed to analyze the results of the research, but there was less research applying various staining methods to cartilage research or exploring staining mechanism.OBJECTIVE: To explore the advantages and disadvantages of rat knee joint with different staining methods.METHODS: Articular cartilage of the normal rat knee joint was sampled. The structure of normal cartilage was observed through the histological staining of hematoxylin-eosin, safranin O, alcian blue, toluidine blue, safranin-alcian blue, as well as safranin-fast green.RESULTS AND CONCLUSION: Tidemarks with blue, red and blue colors could be observed in hematoxylin-eosin, safranin O and toluidine blue staining, respectively. The stain intensity of Safranin O and toluidine blue staining were increased toward tidemark. Thus, Safranin O staining was superior to other methods in observing tidemark. The four-layer structures of cartilage were exhibited clearly, with arranged chondrocytes columnar and basophilic matrix in hematoxylin-eosin staining. The safranin O staining showed that the four-layer structure of cartilage was clear, and the matrix at bottom was stained redder than other places. Alcian blue staining showed that, peripheral chondrocytes were strong stained at pH1.0, which stained deeper at PH2.5. The cartilage structure was not clearly in toluidine blue staining, with clearly nuclei and almost no coloring cytoplasm, but matrix was stained light blue purple. Safranirn-alcian blue staining showed that the cartilage surface and matrix exhibited different red, which was deep red at the bottom zones, but the surrounding of chondrocytes was stained blue. There was obviously contrast between cartilage tissue and bone tissue in safranin O staining, which showed red in the former and green in the latter. The results demonstrated that the 4-layer structure of cartilage can be observed by all above methods, especially safranin O staining, which is excellent in observing cartilage layers and tidemark structures. Hematoxylin-eosin staining provides better outcomes than other methods in observing morphologic changes of chondrocyte.