Background
Methods
Chondrocyte isolation and culture
Cytokine quantification by ELISA
Quantification of nitrites
RNA purification
Reverse Transcription Real-time Polymerase Chain Reaction (Real-time RT-PCR) analysis
Gene name | Gene symbol | Assay ID |
---|---|---|
IL6 | interleukin 6 | Hs00174131_m1 |
IL8 | interleukin 8 | Hs00174103_m1 |
MMP3 | matrix metalloproteinase 3 | Hs00968305_m1 |
MMP13 | matrix metalloproteinase 13 | Hs00233992_m1 |
TIMP1 | TIMP metallopeptidase inhibitor 1 | Hs00171558_m1 |
TIMP3 | TIMP metallopeptidase inhibitor 3 | Hs00165949_m1 |
TIMP4 | TIMP metallopeptidase inhibitor 4 | Hs00162784_m1 |
ACTB | actin, beta | Hs99999903_m1 |
GAPDH | glyceraldehyde-3-phosphate dehydrogenase | Hs99999905_m1 |
RPLP0 | ribosomal protein, large, P0 | Hs99999902_m1 |
HPRT1 | hypoxanthine phosphoribosyltransferase 1 | Hs99999909_m1 |
Statistical analysis
Results
Effect of pro-inflammatory cytokines on human chondrocytes
Stimulus | |||
---|---|---|---|
Molecules in the Supernatants (ng/mL) | Medium | IL-1β | TNF-α |
IL-6 | 3.0 ± 0.2 | 367 ± 4.9* | 70 ± 3.7* |
IL-8 | 0.0 | 781 ± 32* | 120 ± 5.6* |
MMP-3 | 2.0 ± 0.00 | 14500 ± 400* | 8100 ± 50* |
MMP-13 | 8.1 ± 0.5 | 71.0 ± 8.0* | 35.3 ± 2.7* |
TIMP-3 | 1.63 ± 0.12 | 2.71 ± 0.05° | 1.23 ± 0.01 ns |
TIMP-4 | 0.11 ± 0.01 | 0.12 ± 0.01 ns | 0.12 ± 0.01 ns |
NO (μM) | 19.5 ± 3.5 | 102.8 ± 32.6* | 95.8 ± 36.9* |
Stimulus | ||
---|---|---|
Relative gene expression for | IL-1β | TNF-α |
IL-6 | 2767 ± 176* | 680 ± 49* |
IL-8 | 5090 ± 271* | 1088 ± 78* |
MMP-3 | 112 ± 10.0* | 71 ± 6.5* |
MMP-13 | 7.5 ± 2.0* | 6.5 ± 0.5* |
TIMP-3 | 3.4 ± 0.7° | 1.0 ± 0.1 ns |
TIMP-4 | 1.9 ± 0.2° | 1.0 ± 0.05 ns |
iNOS | 1656 ± 100* | 1663 ± 110* |