Histological analysis

Tissue samples were decalcified and processed for histology studies. Paraffin sections were stained with haematoxylin and eosin (H&E) (Thermo Fisher Scientific, Scoresby, Australia) to confirm the histological structure of the arthritic hind paws and to study the various features for morphology. Sections were also evaluated for cartilage proteoglycan depletion with Toluidine Blue (Sigma-Aldrich, Sydney, Australia). Histological sections were scored from 0 to 3 (0 none, 1 mild, 2 moderate, and 3 severe) by a “blinded” observer for five parameters: (i) synovitis, (ii) joint space exudate, (iii) soft tissue inflammation, (iv) cartilage degradation, and (v) bone damage. The maximum obtainable score was 15.