Assessing the Wound Healing Efficacy of Wrightia tinctoria Protease-Hydrolysed Cobia Skin Collagen: An In Vivo and In Silico Study.

Wrightia tinctoria crude extract-hydrolysed cobia skin collagen (WHC) was evaluated for its wound healing potential through in vivo and in silico approaches. WHC was assessed in a murine excisional wound model, and mechanistic insights were explored using protein-protein interaction network analysis and molecular docking studies. Topical application of WHC resulted in complete wound closure within 10 days, compared with 14 days in the vehicle control. Histopathological examination demonstrated effective restoration of epidermal and dermal architecture in healed tissues. Immunohistochemical analyses confirmed re-establishment of epidermal differentiation, evidenced by keratin 1 and keratin 5 expression and collagen type I fibrillar organisation, while enhanced CD31 staining indicated increased angiogenesis in WHC-treated wounds. LC-MS/MS analysis identified four collagen-derived peptides identical to native cobia collagen sequences, with molecular weights ranging from 1.6 to 2.6 kDa and peptide lengths of 18-30 amino acids. In silico network analysis suggested interactions of collagen type I with integrin receptors, and molecular docking results predicted binding affinities of the identified peptides towards integrin α2β1, surpassing that of the positive control. Collectively, these findings demonstrate the therapeutic potential of W. tinctoria-hydrolysed cobia skin collagen as a promising bioactive material for wound management.