Comparative Evaluation of Liposome Encapsulated Hygrophila auriculata (Schumach.) Heine Root and Betulin on Wound Healing Activity through In silico, In vitro and In vivo approaches

Abstract

The present study was conducted to evaluate the comparative wound healing activity newlineassociated with Liposome-encapsulated Hygrophila auriculata (K. Schum.) Heine Root and Betulin. The research was done in four phases. Initially, studies on phytochemicals and free radical scavenging activities were carried out on various extracts from H. auriculata leaves and roots. Subsequently, in silico studies were conducted to identify lead compounds against wound targets from secondary metabolites of H. auriculata roots. Phase III involved the synthesis and characterization of liposomes using EEHA (Ethanol Extract of H. auriculata) and betulin. In the final phase, the in vitro and in vivo wound healing potential of the newlinesynthesized liposomes was assessed using the human keratinocyte cell line, HACAT, and newlineswiss albino rats. newlineThe qualitative phytochemical analysis revealed the presence of eight major newlinephytoconstituents: alkaloids, phenols, tannins, flavonoids, proteins, carbohydrates, cardiac glycosides, and terpenoids. The ethanol extract of H. auriculata demonstrated significant scavenging effects on DPPH, FRAP, and ABTS, compared to other solvent extracts. Due to its superior phytochemical content and free radical scavenging capability, the ethanol extract was selected for further investigation. Enzymic antioxidants (catalase, superoxide dismutase, newlineperoxidase, polyphenol oxidase, and glutathione S transferase) and non-enzymic antioxidants (and#945;-tocopherol, polyphenols, and ascorbic acid) were evaluated in the ethanol extract. Notably, the roots exhibited the highest concentration of antioxidants, prompting the selection of roots for subsequent studies. LC-MS analysis of H. auriculata root identified 15 significant newlinecompounds, and their physicochemical properties were examined. Ten ligands were analyzed for docking characteristics with proteins crucial for wound healing prognosis. Among the top-scoring complexes, since betulin exhibited the highest affinity, LB (Liposome encapsulated betulin) was synthesized and further evaluated