Integrated in vitro and in silico evaluation of rutin as a potential modulator of wound healing pathways

Wound healing is a multifactorial biological process that requires the coordinated regulation of inflammation, cell migration, angiogenesis, growth, and extracellular matrix remodeling. These drawbacks of existing wound care treatments prompt the need to seek safe and cost-effective agents and multi-target agents. This was demonstrated in a study that investigated the wound-healing capacity of a natural flavonoid glycoside, rutin, using an integrated in vitro-in silico methodology. An MTT assay was used to determine the cytocompatibility of rutin in L929 fibroblast cells, and the results showed that cell viability was high at a wide range of concentrations. A scratch wound healing assay showed enhanced fibroblast migration and wound closure in a concentration-dependent manner, and wound contraction was nearly complete within 72 h at optimum concentrations. To understand the molecular mechanisms underlying these effects, network pharmacological analysis revealed 94 common targets between rutin-associated wound healing and rutin-associated genes. Protein-protein interaction analysis identified key regulatory nodes, such as PIK3R1, PRKCA, and EGFR, which are central to the pathways that regulate cell proliferation, migration, angiogenesis, and inflammatory regulation. Enrichment analysis of gene ontology and KEGG pathways revealed that the PI3K-Akt, MAPK, VEGF, and AGE-RAGE signaling pathways were highly involved. Molecular docking showed good binding affinities of rutin to PIK3R1, PRKCA, and EGFR, which was also confirmed by 100 ns molecular dynamics simulations that revealed complex stability and favorable conformational behavior. Density functional theory analysis showed that the electronic characteristics were in line with the antioxidant activity and the strong intermolecular interactions. Together, these results demonstrate that rutin is a promising multifunctional topical wound-healing agent and provide mechanistic evidence for its use in topical therapeutic applications.