Tissue samples were evaluated for indirect immunostaining with the mouse monoclonal antibodies Col2A1 (M2139) and MMP-13 (C-3) from Santa Cruz Biotechnology, using a biotinylated anti-mouse secondary antibody (Donkey anti-mouse IgG (H + L) Biotin, Millipore, Burlington, MA, USA) and the avidin–biotin complex (ABC Elite PK-6100, Vector, Burlingame, CA, USA). Antibody binding was revealed with the DAB-peroxidase kit (Peroxidase Substrate Kit DAB SK-4100 Vector) and counterstaining with Crystal Violet (C5042, Sigma, St. Louis, MO, USA).
The presence of type II collagen (Col2A1) is related to the presence of healthy tissue, which, when hypertrophied after damage, begins to be replaced by type X collagen. On the other hand, metalloproteinase 13 (MMP- 13) is the most widely used marker of inflammation in the evaluation of cartilage, as it is the main protease in articular cartilage degradation [21 ,22 (link)].
The presence of type II collagen (Col2A1) is related to the presence of healthy tissue, which, when hypertrophied after damage, begins to be replaced by type X collagen. On the other hand, metalloproteinase 13 (MMP- 13) is the most widely used marker of inflammation in the evaluation of cartilage, as it is the main protease in articular cartilage degradation [21 ,22 (link)].