Brown M P, Trumble T N, Sandy J D, Merritt K A
Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610-1432, USA.
Osteoarthritis Cartilage. 2007 Dec;15(12):1443-5. doi: 10.1016/j.joca.2007.05.018. Epub 2007 Jul 16.
To 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).
SF 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.
DMMB analysis showed that SF CS peak chain length in the OC injury group increased significantly (18.7 kDa) when compared to rested and exercised normal horses (15.6 kDa). The assay had a positive predictive value of 71% and a negative predictive value of 75% for discriminating between normal and injured joints.
We 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.
确定二甲基亚甲基蓝（DMMB）分析结合琼脂糖凝胶过滤色谱法（Superose 6）是否可替代荧光团辅助碳水化合物电泳（FACE）来测定滑液（SF）中硫酸软骨素（CS）的链长。
从以下三组马匹获取滑液：（1）休息8周后的正常马匹；（2）跑步机训练9个月后的同一批马匹；（3）赛马中患有骨软骨（OC）损伤的马匹。通过凝胶色谱法和DMMB分析测定滑液CS浓度和链长，并与之前对相同样本进行FACE分析得到的结果进行比较。
DMMB分析显示，与休息和运动后的正常马匹（15.6 kDa）相比，OC损伤组滑液CS的峰值链长显著增加（18.7 kDa）。该检测方法区分正常关节和损伤关节的阳性预测值为71%，阴性预测值为75%。
我们报告了一种简单且廉价的DMMB分析滑液CS链长的方法，该方法结合Superose 6色谱法可区分正常关节和损伤后关节。与我们之前的FACE分析结果[布朗MP、特朗布尔TN、普拉斯AHK、桑迪JD、罗曼诺M、埃尔南德斯J等人。运动和损伤会增加滑液中硫酸软骨素链长并缩短透明质酸链长。骨关节炎与软骨2007；15]相似，我们的DMMB结果显示损伤关节滑液中CS的链长增加。
