Osteochondral injury increases type II collagen degradation products (C2C) in synovial fluid of Thoroughbred racehorses

SummaryObjectivesTo investigate the effects of exercise and osteochondral (OC) injury on type II collagen degradation products (collagenase cleavage neoepitope commercially known as C2C) in synovial fluid (SF) from Thoroughbred (TB) racehorses and to compare these results with radiographic and arthroscopic scores of severity of joint injury.MethodsMetacarpophalangeal/metatarsophalangeal (MCP/MTP) and carpal SF was obtained from (1) 20 normal rested horses, (2) the same horses after 5 to 6 months of race training, and (3) 27 horses with OC injury from racing. For group 3, radiographic and arthroscopic scores were determined. Concentrations of SF C2C were determined by ELISA.ResultsSF C2C concentrations in OC injured carpal and MCP/MTP joints were significantly different than rested and exercised joints (P<0.01). However, carpal and MCP/MTP SF C2C concentrations were not significantly different between rested and exercised groups. Arthroscopic scores were significantly higher for OC injured carpal than OC injured MCP/MTP joints (P=0.002). OC injured SF C2C concentrations were positively correlated with radiographic and arthroscopic scores. Arthroscopic scores were positively correlated with radiographic scores. SF C2C concentrations ≥64pmol/mL for MCP/MTP joints and ≥75pmol/mL for carpal joints discriminated OC injured joints from rested or exercised joints.ConclusionOC injury caused a significant increase in SF C2C concentrations in carpal and MCP/MTP joints compared to rested and exercised horses. SF C2C concentrations were correlated to severity of joint injury. Based on these findings, SF C2C analysis may be useful for evaluation of joint injury

A simplified method of determining synovial fluid chondroitin sulfate chain length

SummaryObjectiveTo determine whether dimethylmethylene blue (DMMB) analysis, when combined with agarose gel filtration chromatography (Superose 6), can be performed instead of fluorophore-assisted carbohydrate electrophoresis (FACE) to determine chondroitin sulfate (CS) chain length in synovial fluid (SF).MethodsSF was obtained from (1) normal horses after 8 weeks of rest, (2) the same horses after 9 months of treadmill training, and (3) horses with osteochondral (OC) injury from racing. SF CS concentrations and chain lengths were determined by gel chromatography and DMMB analysis and compared with previous results determined by FACE analysis on the same samples.ResultsDMMB analysis showed that SF CS peak chain length in the OC injury group increased significantly (18.7kDa) when compared to rested and exercised normal horses (15.6kDa). The assay had a positive predictive value of 71% and a negative predictive value of 75% for discriminating between normal and injured joints.ConclusionsWe report a simple and inexpensive DMMB analysis of SF CS chain length, which, when coupled with Superose 6 chromatography, discriminates between normal and post-injury joints. Similar to our previous FACE analysis results [Brown MP, Trumble TN, Plaas AHK, Sandy JD, Romano M, Hernandez J, et-al. Exercise and injury increase chondroitin sulfate chain length and decrease hyaluronan chain length in synovial fluid. Osteoarthritis Cartilage 2007;15], our DMMB results show an increase in the chain length of the CS in the SF of injured joints