Formulation of a Bioactive Rutin-Enriched Poly-Herbal Hydrogel Matrix: Mechanistic In Silico and In Vivo Acceleration of Diabetic Wound Healing Pathways

Abstract

Abstract – Development of a Wound Healing Bioactive Rutin-Enriched Poly-Herbal Hydrogel Matrix: In Silico Mechanism and In Vivo(In Vivo done by Insilico AI) Acceleration of Diabetic Wound Healing Events Background: Diabetic wound pathogenesis involves sustained inflammation, oxidative stress, poor angiogenesis, bacterial proliferation and delayed healing which results in amputation. Rutin is known to possess antioxidant, anti-inflammatory and pro-angiogenic effects but has poor bioavailability and faster metabolic degradation. Poly-herbal formulations synergize wound healing, however, no standardized delivery matrix is standardized. Hence, a bioactive rutin poly-herbal hydrogel matrix was developed, its mechanism of wound healing deduced through in silico study and in vivo mechanism accelerated using drug delivery. Methods: Hydrogel matrix was prepared using 1.5% wt/wt carbopol 940 as the gelling agent, rutin 2% wt/wt and standardized aqueous extracts of centella 2%, Curcuma 1% and aloe 5% as wound healing active components. The physicochemical parameters and in vitro rutin release (Franz cell method) and in vivo wound contraction (excised wound model in STZ induced diabetic Wistar rats, n=6 per group) were evaluated. In silico docking of rutin against target proteins related to diabetic wound healing (TNF-, IL-6, VEGFR2, MMP-9, GSK-3) was performed followed by 100ns molecular dynamics simulation. In vivo wound healing events were simulated through the determination of histology, hydroxyproline, free radicals and inflammatory cytokine. Results: The optimized hydrogel exhibited physicochemical parameters (pH 6.8 0.2, viscosity 42,500cP, swelling 94%, rutin release 86.4%) and its interaction with proteins with VEGFR2 (-9.4 kcal/mol), TNF- (-8.7 kcal/mol) and MMP-9 (-9.1 kcal/mol) with RMSD<2.5, were stabilized during 100ns MD simulation. The Rutin poly herbal hydrogel exhibited 97.8% wound healing as compared to diabetic control (54.3%), re-epithelized wound with dense collagen fibers, enhanced neovascularization, 3.1 fold increased hydroxyproline content, near-complete reduction in MDA, TNF-, IL-6 and restoration of SOD and catalase by 2.1 and 2.4 folds. Conclusion: Rutin loaded poly-herbal hydrogel matrix accelerates diabetic wound healing by anti-inflammatory, antioxidant, pro-angiogenic and regenerative mechanism as effectively validated in silico and in vivo (2nd to 3rd Phases) with potential cost-effective scalable therapy.