Bayliss M T, Venn M, Maroudas A, Ali S Y
Biochem J. 1983 Feb 1;209(2):387-400. doi: 10.1042/bj2090387.
Full-depth plugs of adult human articular cartilage were cut into serial slices from the articular surface and analysed for their glycosaminoglycan content. The amount of chondroitin sulphate was highest in the mid-zone, whereas keratan sulphate increased progressively through the depth. Proteoglycans were isolated from each layer by extraction with 4M-guanidinium chloride followed by centrifugation in 0.4M-guanidinium chloride/CsCl at a starting density of 1.5 g/ml. The efficiency with which proteoglycans were extracted depended on slice thickness, and extraction was complete only when cartilage from each zone was sectioned at 20 microns or less. When thick sections (250 microns) were extracted, hyaluronic acid was retained in the tissue. Most of the proteoglycans, extracted from each layer under optimum conditions, could interact with hyaluronic acid to form aggregates, although the extent of aggregation was less in the deeper layers. Two pools of proteoglycan were identified in all layers by gel chromatography (Kav. 0.33 and 0.58). The smaller of these was rich in keratan sulphate and protein, and gradually increased in proportion through the cartilage depth. Chondroitin sulphate chain size was constant in all regions. The changes in composition and structure observed were consistent with the current model for hyaline-cartilage proteoglycans and were similar to those observed with increasing age in human articular cartilage.
将成人关节软骨的全层塞从关节表面切成连续切片，并分析其糖胺聚糖含量。硫酸软骨素的含量在中间区最高，而硫酸角质素则随深度逐渐增加。通过用4M氯化胍提取，然后在起始密度为1.5g/ml的0.4M氯化胍/CsCl中离心，从每一层中分离蛋白聚糖。蛋白聚糖的提取效率取决于切片厚度，只有当每个区域的软骨切成20微米或更薄时，提取才会完全。当提取厚切片（250微米）时，透明质酸保留在组织中。在最佳条件下从每一层提取的大多数蛋白聚糖能够与透明质酸相互作用形成聚集体，尽管在较深层聚集体的程度较小。通过凝胶色谱法（Kav.0.33和0.58）在所有层中鉴定出两个蛋白聚糖池。其中较小的富含硫酸角质素和蛋白质，并在软骨深度中比例逐渐增加。硫酸软骨素链大小在所有区域都是恒定的。观察到的组成和结构变化与目前的透明软骨蛋白聚糖模型一致，并且与在人类关节软骨中随年龄增长观察到的变化相似。
