Natural multi-target inhibitors from Adhatoda vasica and Calotropis procera for dermal regeneration.

Natural plant-derived compounds are increasingly explored as alternatives for managing skin disorders associated with oxidative stress, enzymatic degradation, and fibroblast dysfunction. In this study, bioactive constituents from Adhatoda vasica and Calotropis procera were isolated and characterized using UF-HPLC-DAD and Fourier-transform infrared (FTIR) spectroscopy. Identified compounds included viscic acid, pasakbumin C, 8'Z-enyl congeners, 5-(8'Z,11'Z-heptadecadienyl)-1,3-benzenediol, 9'-(o-methyl) protocetraric acid, and calophynic acid. The biological activities of leaf and seed extracts were evaluated through in vitro enzymatic assays targeting tyrosinase (monophenolase and diphenolase), collagenase, elastase, and hyaluronidase. A. vasica leaf extracts exhibited notable tyrosinase inhibition, with IC₅₀ values of 22.16 µg/mL for monophenolase and 15.5 µg/mL for diphenolase, while C. procera also demonstrated significant inhibitory activity. Both extracts showed appreciable anti-collagenase, anti-elastase, anti-hyaluronidase, and antioxidant activities, suggesting protective potential against extracellular matrix degradation and oxidative damage. Cytotoxicity assessment using LDH release assays confirmed concentration-dependent effects, and ultrastructural analyses using scanning and transmission electron microscopy revealed extract-induced morphological changes in dermal fibroblasts, including alterations in cell wall integrity, cytoplasmic organization, and intercellular structure. The results suggest that A. vasica and C. procera are promising sources of bioactive compounds with potential applications in dermatological formulations, including topical preparations to manage skin disorders and prevent enzymatic skin degradation.