Example 1
We analysed candidate peptides from region A for their OA phenotype suppressive actions on primary OA articular chondrocytes in the presence of 20% (v/v) OA synovial fluid (SF). Data were combined from three individual OA chondrocyte isolates (n=3) that were each tested in triplicate. Seeded passage-2 OA chondrocytes were exposed to 100 nM of peptide in the absence or presence of OA synovial fluid and after 24 hours analysed for mRNA expression of genes COL10A1, ALP, RUNX2, COL2A1, ACAN, SOX9, COX-2, IL-6, PGE, MMP13 and ADAMTS5 (corrected for 28S rRNA expression and relative to control conditions (no peptide exposure)).
PGE2 secretion in culture supernatant was analysed by EIA and ALP enzyme activity was determined by an in-house developed colorimetric assay for ALP activity. GAG content was determined by colorimetric alcian blue assay.
Results were scored as + or − wherein + means that the peptide decreased the activity of genes COL10A1, ALP, RUNX2, COX-2, IL-6, PGE, MMP13 and ADAMTS5 or increased the activity of COL2A1, ACAN and SOX9. For ALP activity a + means a decreased activity and for GAG activity, a + means an increased activity,
