title: In vitro culture of enzymatically isolated chondrons: a possible model for the initiation of osteoarthritis.
abstract: The aim of this study was to assess whether enzymatically isolated chondrons from normal adult articular cartilage could be used as a model for the onset of osteoarthritis, by comparison with mechanically extracted chondrons from osteoarthritic cartilage. Enzymatically isolated chondrons (EC) were cultured for 4 weeks in alginate beads and agarose gel constructs. Samples were collected at days 1 and 2, and weekly thereafter. Samples were immunolabelled for types II and VI collagen, keratan sulphate and fibronectin and imaged using confocal microscopy. Mechanically extracted chondrons (MC) were isolated, immunohistochemically stained for type VI collagen and examined by confocal microscopy. In culture, EC showed the following characteristics: swelling of the chondron capsule, cell division within the capsule and remodelling of the pericellular microenvironment. This was followed by chondrocyte migration through gaps in the chondron capsule. Four types of cell clusters formed over time in both alginate beads and agarose constructs. Cells within clusters exhibited quite distinct morphologies and also differed in their patterns of matrix deposition. These differences in behaviour may be due to the origin of the chondrocytes in the intact tissue. The behaviour of EC in culture paralleled the range of morphologies observed in MC, which presented as single and double chondrons and large chondron clusters. This preliminary study indicates that EC in culture share similar structural characteristics with MC isolated from osteoarthritic cartilage, confirming that some processes that occur in osteoarthritis, such as pericellular remodelling, take place in EC cultures. The study of EC in culture may therefore provide an additional tool to investigate the mechanisms operating during the initial stages of osteoarthritis. Further investigation of specific osteoarthritic phenotype markers will, however, be required in order to validate the value of this model.
authors.affiliation: Department of Anatomy with Radiology, Faculty of Medical and Health Sciences, The University of Auckland, New Zealand. jacqui.ross@auckland.ac.nz
authors.forename: J M
authors.initials: JM
authors.lastname: Ross
authors.forename: A F
authors.initials: AF
authors.lastname: Sherwin
authors.forename: C A
authors.initials: CA
authors.lastname: Poole
mesh_terms: D000464:Alginates; D000818:Animals; D002358:Cartilage, Articular / Q000473:pathology*; D002465:Cell Movement; D019902:Chondrocytes / Q000473:pathology*; D024043:Collagen Type II / Q000032:analysis; D024141:Collagen Type IV / Q000032:analysis; D004285:Dogs; D005353:Fibronectins / Q000032:analysis; D005782:Gels; D007150:Immunohistochemistry; D007632:Keratan Sulfate / Q000032:analysis; D018613:Microscopy, Confocal; D008863:Microspheres; D023421:Models, Animal; D010003:Osteoarthritis / Q000473:pathology*; D012685:Sepharose; D046509:Tissue Culture Techniques
chemical_list: D000464:Alginates; D024043:Collagen Type II; D024141:Collagen Type IV; D005353:Fibronectins; D005782:Gels; D012685:Sepharose; D007632:Keratan Sulfate