Unlocking the wound healing potential of Anastatica hierochuntica L. (Kaff Maryam) Extract's nanosuspension: UPLC-MS/MS metabolic profiling, formulation development and statistical optimization

Anastatica hierochuntica L. (Kaff Maryam) is a desert plant with documented therapeutic applications. Various medicinal uses have been reported for the plant extract such as antioxidant, anti-inflammatory, antifungal, antimicrobial, hypoglycemic, hypolipidemic, and hepatoprotective activities. The abundant plants constituents are promising and encourage the investigation of novel therapeutic uses. The principal objective of this study was to identify the bioactive constituents of A. hierochuntica (AH) by phytochemical profiling of its ethanolic extract, alongside an investigation of its wound healing efficacy after being formulated into nanosuspension for topical localized wound treatment. A comprehensive metabolomics study of the ethanolic extract of A. hierochuntica aerial parts was assessed using UPLC MS/MS technique. Additionally, the preparation of AH nanosuspension was executed utilizing the antisolvent-precipitation method. A D-optimal experimental design was adopted to investigate the influence of variables such as: stabilizer ratio, antisolvent: solvent ratio, and stabilizer type on particle size (PS), polydispersity index (PDI), and zeta potential (ZP). The design also was employed in the optimization process. The selected optimal formula was further assessed for size characteristics, morphological identification by TEM, storage stability, FTIR, cytocompatibility with human skin fibroblast (HSF), and scratch assay on HSF to assess wound healing aptitude. Chemical profiling of the aerial parts revealed an abundance of glucosinolates, flavonoids, flavolignans, and fatty acids. The optimized AH-nanosuspension showed small particle size (128.87 ± 26 nm), small PDI (0.136 ± 0.05), and high ZP absolute value (−37.53 ± 0.52 mV). Furthermore, TEM photography of the optimal nanosuspension formula exhibited circular non-aggregating nanosized particles. FTIR confirmed the successful formulation of the AH ethanolic extract into nanosuspension with no physicochemical interactions. Additionally, AH-nanosuspension demonstrated cytocompatibility with HSF with IC50 = 58.317 μg/mL. Moreover, both AH pure extract and AH-nanosuspension exhibited a remarkable cell migration rate and wound closure % which propounds an auspicious wound healing potential. The study accentuates the supremacy of the nanosized delivery approach of AH aerial parts ethanolic extract and warrants further in-vivo investigations to establish its clinical utility for wound management. [Display o