Shohani Behnaz, Orazizadeh Mahmoud, Hashemitabar Mahmoud, Heinegard Dick
Cellular and Molecular Research Center, Dept. of Anatomical Sciences, Faculty of Medicine, Ahvaz Jundishapour University of medical Sciences, Ahvaz, Iran.
Dept. of Clinical Sciences, Section for Molecular Skeletal Biology, BMC, Plan C12, Lund, Sweden.
Iran Biomed J. 2010 Oct;14(4):158-63.
This work aimed to show and compare the degradation time of some of cartilage extracellular matrix components using an in vitro model for cartilage degradation induced by interleukin-1α. It is known that elucidation of molecular events under Interleukin-1α induction of bovine nasal cartilage could obtain useful data to understand more about involving mechanisms for tissue breakdown in joint disease.
The cartilage was taken from an adult bovine in a local slaughterhouse. After removing the whole perichondrium, the equal 2 mm diameter pieces of bovine nasal cartilage were punched out and cultured in Dulbecco's modified Eagle's medium DMEM with or without 10 ng/ml Interleukin-1α for 24 days. Each 3 days, the media were removed and exchanged by fresh media, and the removed media were stored in -20 degrees C. Sodium dodecyl sulphate/polyacrylamid-gel electrophoresis SDS-PAGE and Western-blot methods were used for analyzing the samples.
The first fragment of fibromodulin (FM) was seen at day 6 and further fragments were appeared at day 18. Cartilage oligomeric matrix protein (COMP) releasing was as a successive pattern during culture period and the first fragment was found at day 6. Collagen IX fragments were seen at day 9 and in a progressive pattern until the end of the study.
This study shows that FM and COMP could be considered as the suitable candidates for studying the mechanisms that participate in the cartilage degradations.
本研究旨在利用白细胞介素-1α诱导软骨降解的体外模型，展示并比较软骨细胞外基质某些成分的降解时间。众所周知，阐明白细胞介素-1α诱导牛鼻软骨过程中的分子事件，可为深入了解关节疾病中组织破坏的相关机制提供有用数据。
软骨取自当地屠宰场的成年牛。去除整个软骨膜后，冲出直径相等的2毫米牛鼻软骨片，在含或不含10纳克/毫升白细胞介素-1α的杜尔贝科改良伊格尔培养基（DMEM）中培养24天。每3天更换一次培养基，更换出的培养基储存于-20℃。采用十二烷基硫酸钠/聚丙烯酰胺凝胶电泳（SDS-PAGE）和蛋白质免疫印迹法分析样本。
纤调蛋白（FM）的首个片段在第6天出现，后续片段在第18天出现。软骨寡聚基质蛋白（COMP）在培养期间呈连续释放模式，首个片段在第6天发现。IX型胶原片段在第9天出现，并呈渐进模式直至研究结束。
本研究表明，FM和COMP可被视为研究参与软骨降解机制的合适候选物。
