Interleukin-1beta induces different gene expression of stromelysin, aggrecan and tumor-necrosis-factor-stimulated gene 6 in human osteoarthritic chondrocytes in vitro.
OBJECTIVE
To analyze the gene expression of osteoarthritic chondrocytes cultured in alginate after stimulation with interleukin (IL)-1beta.


METHODS
Chondrocytes were isolated from osteoarthritic cartilage obtained during total knee replacement by sequential enzymatic digestion. After suspension in alginate, cells were cultured with and without 100 pg/ml IL-1beta. Quantitative RT-PCR reaction was used to estimate the messenger RNA (mRNA) of three different metabolites [tumor-necrosis-factor-stimulated gene 6 (TSG-6), stromelysin-1 (MMP-3) and aggrecan (AGG)].


RESULTS
After having shown the precision of quantitative PCR, this method allowed us to detect IL-1beta-induced changes in mRNA of TSG-6, MMP-3 and AGG. MMP-3 was found to be the most abundant transcript, IL-1beta induced a 12-fold upregulation of MMP-3 levels compared to control, and 7-fold of TSG-6. The AGG transcript level, indicating anabolic events, was found to be downregulated by between 2- and 3-fold.


CONCLUSIONS
In our culture system, the response of osteoarthritic chondrocytes to IL-1beta is preserved. Therefore, this system might be helpful for further investigation of the influences of drugs, cytokines and growth factors, for example, on the metabolism of chondrocytes at the level of gene transcription as the most basic level of regulation.
Stöve
J
J
Department of Orthopaedic Surgery (RKU), University of Ulm, Oberer Eselsberg 45, D-89081 Ulm, Germany. johannes.stoeve@medizin.uni-ulm.de
Huch
K
K
Günther
K P
KP
Scharf
H P
HP
D000368:Aged; D053674:Aggrecans; D002358:Cartilage, Articular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000235:genetics* / Q000378:metabolism; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects* / Q000378:metabolism; D004247:DNA / Q000032:analysis; D017931:DNA Primers / Q000737:chemistry; D016326:Extracellular Matrix Proteins; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology*; D037181:Lectins, C-Type; D008297:Male; D019278:Matrix Metalloproteinase 3 / Q000235:genetics* / Q000378:metabolism; D020370:Osteoarthritis, Knee / Q000378:metabolism* / Q000473:pathology; D011509:Proteoglycans / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction
D053674:Aggrecans; D015815:Cell Adhesion Molecules; D017931:DNA Primers; D016326:Extracellular Matrix Proteins; D007375:Interleukin-1; D037181:Lectins, C-Type; D011509:Proteoglycans; D012333:RNA, Messenger; C072273:TNFAIP6 protein, human; D004247:DNA; D019278:Matrix Metalloproteinase 3