Melrose James, Smith Susan, Cake Martin, Read Richard, Whitelock John
Raymond Purves Bone and Joint Research Laboratories, Institute of Bone and Joint Research, Level 5, The University Clinic, Building B26, The Royal North Shore Hospital, St. Leonards, NSW, 2065, Australia.
Histochem Cell Biol. 2005 Sep;124(3-4):225-35. doi: 10.1007/s00418-005-0005-0. Epub 2005 Oct 28.
This is the first study to immunolocalise perlecan in meniscal tissues and to demonstrate how its localisation varied with ageing relative to aggrecan and type I, II and IV collagen. Perlecan was present in the middle and inner meniscal zones where it was expressed by cells of an oval or rounded morphology. Unlike the other components visualised in this study, perlecan was strongly cell associated and its levels fell significantly with age onset and cell number decline. The peripheral outer meniscal zones displayed very little perlecan staining other than in small blood vessels. Picrosirius red staining viewed under polarised light strongly delineated complex arrangements of slender discrete randomly oriented collagen fibre bundles as well as transverse, thick, strongly oriented, collagen tie bundles in the middle and outer meniscal zones. The collagen fibres demarcated areas of the meniscus which were rich in anionic toluidine blue positive proteoglycans; immunolocalisations confirmed the presence of aggrecan and perlecan. When meniscal sections were examined macroscopically, type II collagen localisation in the inner meniscal zone was readily evident in the 2- to 7-day-old specimens; this became more disperse in the older meniscal specimens. Type I collagen had a widespread distribution in all meniscal zones at all time points. Type IV collagen was strongly associated with blood vessels in the 2- to 7-day-old meniscal specimens but was virtually undetectable at the later time points (>7 month).
这是第一项对半月板组织中的核心蛋白聚糖进行免疫定位,并证明其定位如何随年龄增长相对于聚集蛋白聚糖以及I、II和IV型胶原蛋白而变化的研究。核心蛋白聚糖存在于半月板的中间和内部区域,由椭圆形或圆形形态的细胞表达。与本研究中观察到的其他成分不同,核心蛋白聚糖与细胞紧密相关,并且其水平随着年龄增长和细胞数量减少而显著下降。除了小血管外,半月板外周区域的核心蛋白聚糖染色很少。在偏振光下观察的天狼星红染色清晰地勾勒出半月板中间和外周区域中细长离散随机取向的胶原纤维束以及横向、粗大、强取向的胶原连接束的复杂排列。胶原纤维划分出半月板中富含阴离子甲苯胺蓝阳性蛋白聚糖的区域;免疫定位证实了聚集蛋白聚糖和核心蛋白聚糖的存在。当宏观检查半月板切片时,II型胶原蛋白在2至7日龄标本的半月板内部区域的定位很明显;在较老的半月板标本中,这种定位变得更加分散。I型胶原蛋白在所有时间点的所有半月板区域均有广泛分布。IV型胶原蛋白在2至7日龄的半月板标本中与血管紧密相关,但在后期时间点(>7个月)几乎检测不到。