title: Immunolocalization of collagen types II and III in single fibrils of human articular cartilage.
abstract: Type II and III fibrillar collagens were localized by immunogold electron microscopy in resin sections of human femoral articular cartilage taken from the upper radial zone in specimens from patients with osteoarthritis. Tissue samples stabilized by high-pressure cryofixation were processed by freeze-substitution, either in acetone containing osmium or in methanol without chemical fixatives, before embedding in epoxy or Lowicryl resin, respectively. Ultrastructural preservation was superior with osmium-acetone, although it was not possible to localize collagens by this method. In contrast, in tissue prepared by low-temperature methods without chemical fixation, collagens were successfully localized with mono- or polyclonal antibodies to the helical (Types II and III) and amino-propeptide (Type III procollagen) domains of the molecule. Dual localization using secondary antibodies labeled with 5- or 10-nm gold particles demonstrated the presence of Types II and III collagen associated within single periodic banded fibrils. Collagen fibrils in articular cartilage are understood to be heteropolymers mainly of Types II, IX, and XI collagen. Our observations provide further evidence for the complexity of these assemblies, with the potential for interactions between at least 11 distinct collagen types as well as several noncollagenous components of the extracellular matrix.
authors.affiliation: School of Biosciences, Cardiff University, Cardiff, United Kingdom.
authors.forename: R D
authors.initials: RD
authors.lastname: Young
authors.forename: P A
authors.initials: PA
authors.lastname: Lawrence
authors.forename: V C
authors.initials: VC
authors.lastname: Duance
authors.forename: T
authors.initials: T
authors.lastname: Aigner
authors.forename: P
authors.initials: P
authors.lastname: Monaghan
mesh_terms: D002358:Cartilage, Articular / Q000378:metabolism* / Q000648:ultrastructure; D003094:Collagen / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D020894:Microfibrils / Q000378:metabolism*; D008854:Microscopy, Electron
chemical_list: D003094:Collagen