Co-culture of mechanically injured cartilage with joint capsule tissue alters chondrocyte expression patterns and increases ADAMTS5 production.
We studied changes in chondrocyte gene expression, aggrecan degradation, and aggrecanase production and activity in normal and mechanically injured cartilage co-cultured with joint capsule tissue. Chondrocyte expression of 21 genes was measured at 1, 2, 4, 6, 12, and 24h after treatment; clustering analysis enabled identification of co-expression profiles. Aggrecan fragments retained in cartilage and released to medium and loss of cartilage sGAG were quantified. Increased expression of MMP-13 and ADAMTS4 clustered with effects of co-culture, while increased expression of ADAMTS5, MMP-3, TGF-beta, c-fos, c-jun clustered with cartilage injury. ADAMTS5 protein within cartilage (immunohistochemistry) increased following injury and with co-culture. Cartilage sGAG decreased over 16-days, most severely following injury plus co-culture. Cartilage aggrecan was cleaved at aggrecanase sites in the interglobular and C-terminal domains, resulting in loss of the G3 domain, especially after injury plus co-culture. Together, these results support the hypothesis that interactions between injured cartilage and other joint tissues are important in matrix catabolism after joint injury.
Lee
J H
JH
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Fitzgerald
J B
JB
DiMicco
M A
MA
Cheng
D M
DM
Flannery
C R
CR
Sandy
J D
JD
Plaas
A H
AH
Grodzinsky
A J
AJ
D051722:ADAM Proteins / Q000096:biosynthesis*; D053674:Aggrecans / Q000378:metabolism; D000818:Animals; D002356:Cartilage / Q000293:injuries* / Q000378:metabolism* / Q000473:pathology; D002417:Cattle; D019902:Chondrocytes / Q000378:metabolism* / Q000473:pathology; D018920:Coculture Techniques; D010450:Endopeptidases / Q000378:metabolism; D005786:Gene Expression Regulation; D017746:Joint Capsule / Q000378:metabolism* / Q000473:pathology; D053509:Matrix Metalloproteinase 13 / Q000096:biosynthesis; D017434:Protein Structure, Tertiary; D016760:Proto-Oncogene Proteins c-fos / Q000096:biosynthesis; D016755:Proto-Oncogene Proteins c-jun / Q000096:biosynthesis; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000096:biosynthesis
D053674:Aggrecans; D016760:Proto-Oncogene Proteins c-fos; D016755:Proto-Oncogene Proteins c-jun; D016212:Transforming Growth Factor beta; D010450:Endopeptidases; D051722:ADAM Proteins; D053509:Matrix Metalloproteinase 13; C083641:aggrecanase