Behnaz Shohani1, Mahmoud Orazizadeh1, Mahmoud Hashemitabar1, Dick Heinegard2. 1. Cellular and Molecular Research Center, Dept. of Anatomical Sciences, Faculty of Medicine, Ahvaz Jundishapour University of medical Sciences, Ahvaz, Iran. behnaz.shohani@gmail.com 2. Dept. of Clinical Sciences, zzm321990Section for Molecular Skeletal Biology, BMC, Plan C12, Lund, Sweden.
Abstract
BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSION: This study shows that FM and COMP could be considered as the suitable candidates for studying the mechanisms that participate in the cartilage degradations.
BACKGROUND: 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 bovinenasal cartilage could obtain useful data to understand more about involving mechanisms for tissue breakdown in joint disease. METHODS: The cartilage was taken from an adult bovine in a local slaughterhouse. After removing the whole perichondrium, the equal 2 mm diameter pieces of bovinenasal 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. RESULTS: 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. CONCLUSION: This study shows that FM and COMP could be considered as the suitable candidates for studying the mechanisms that participate in the cartilage degradations.
Authors: B Månsson; D Carey; M Alini; M Ionescu; L C Rosenberg; A R Poole; D Heinegård; T Saxne Journal: J Clin Invest Date: 1995-03 Impact factor: 14.808