Fibroblast Wound Healing Assay

Fibroblast migration was assessed using a “scratch wound” assay, as described previously (13). Fibroblasts were seeded on a six-well plate to a final concentration of 5 × 10⁵ cells/ml. Once confluent, a cell-free area was created with a pipette in each well. Cells were then treated with either TGF-β1 (10 ng/ml) or TGF-β1 along with recombinant hsdc2 (0.5 μg/ml) for 24 hours. Wound closure and fibroblast migration was evaluated at 0, 4, 8, 12, 16, 20, and 24 hours after treatment.