title: Validation of an in vitro single-impact load model of the initiation of osteoarthritis-like changes in articular cartilage.
abstract: The objective of this study was the development and characterization of an in vitro model of the initiation of traumatic osteoarthritis (OA). Articular cartilage was obtained from seven healthy horses and from four horses diagnosed with OA. Cartilage disks were subjected to a single-impact load (500 g from 25, 50, or 100 mm) using a simple drop-tower device and cultured in vitro for up to 20 days. Cartilage sections were examined histologically to observe surface damage and proteoglycan loss. Percentage cell death was determined using TUNEL, release of glycosaminoglycans (GAG) to the medium was measured using the DMMB assay, and percentage weight gain calculated. Following a single-impact load and subsequent culture in vitro, articular cartilage explants demonstrated characteristic surface damage, proteoglycan loss, and chondrocyte death. This closely resembled degenerative changes observed in OA cartilage samples. A kinetic study showed that these degenerative changes (increased weight gain, GAG release into the medium, and chondrocyte death) were initiated within 48 h following impact and increased with recovery time in culture. These parameters were proportional to impact height, that is, impact energy. In conclusion, articular cartilage disks subjected to a single-impact load followed by 48 h of recovery time in culture in vitro developed traumatic OA-like changes. These changes can be quantified and compared, making the in vitro single-impact load model a useful tool for the elucidation of the early molecular pathways involved in the process leading from trauma to cartilage degeneration.
authors.affiliation: Comparative Orthopaedics Research Group, Department of Veterinary Medicine, University of Cambridge, Cambridge, CB3 0ES, United Kingdom. ch353@cam.ac.uk
authors.forename: Camille A M
authors.initials: CA
authors.lastname: Huser
authors.forename: M Elisabeth
authors.initials: ME
authors.lastname: Davies
mesh_terms: D000818:Animals; D017209:Apoptosis; D002358:Cartilage, Articular / Q000473:pathology*; D019902:Chondrocytes / Q000473:pathology; D024043:Collagen Type II / Q000032:analysis; D004195:Disease Models, Animal; D006736:Horses; D020287:In Situ Nick-End Labeling; D010003:Osteoarthritis / Q000473:pathology*; D011509:Proteoglycans / Q000378:metabolism; D015430:Weight Gain
chemical_list: D024043:Collagen Type II; D011509:Proteoglycans