Spatial organization of types I and II collagen in the canine meniscus

Abstract

The meniscus of the knee joint is a fibrocartilage mainly composed of type I collagen and smaller amounts of type II collagen. The distribution of type II collagen in the canine meniscus and its spatial relationship to type I collagen was examined by immunohistochemistry and confocal microscopy. Dorsal and coronal slices of the mid-section of medial and lateral menisci from the knee joints of skeletally mature dogs were predigested with Streptomyces hyaluronate lyase and bacterial Protease enzyme XXIV. Monoclonal antibodies against type I collagen (CP17L) and type II collagen (II-II6B3) and an anti-type II collagen polyclonal antibody (AB759) were employed. The staining for type II collagen in the extracellular matrix of hyaline articular cartilage was diffuse without any identifiable spatial organization. In striking contrast, type II collagen in the fibrocartilage of the meniscus stained as an organized network. Type II collagen was distributed throughout the meniscus with the exception of the outer zone containing the blood vessels. Coronal and dorsal staining of the meniscus showed bundles of circumferential fibrils of type I that colocalized with type II collagen in specific sites. These bundles were enwrapped in a second organizational fibrillar system of types I and II collagen that also colocalized. Bundles of circumferential fibrils appeared in cross-section in coronal sections as dots within the interstitial spaces framed by the network of types I and II collagen of the second system. Confocal overlays showed that types I and II collagens were superimposed, suggesting a close spatial proximity between the two collagens. The cells were confined to the types I and II collagen fibrils that enwrapped the bundles. A striking feature of the radial tie fibers was patches of type II collagen without colocalized type I collagen. Our study reveals a unique network of type II collagen in fibrocartilage of the meniscus that serves as a morphological distinction between fibro- and hyaline cartilage.