Phytochemical profiling of Rosmarinus officinalis aerial parts and exploring its in vitro wound healing activity and network pharmacology

The present study thoroughly assessed the wound healing efficacy of fractions derived from Rosmarinus officinalis through phytochemical profiling, antioxidant assays, and in vitro scratch wound models, along with network pharmacology to identify target genes. HPLC/MS analysis identified rosmarinic acid as the predominant phenolic compound, alongside diterpenoids (carnosic acid, carnosol) and flavonoids (cirsimaritin, diosmetin). The total extract exhibited the highest total phenolic content (106.56 µg gallic acid eq/mg), while the ethyl acetate fraction (ROE) contained the highest flavonoids (43.7 µg rutin eq/mg). Antioxidant assays revealed fraction-dependent efficacy: the n-butanol fraction (RON) showed superior (Ferric Reducing Antioxidant Power) FRAP activity (637.727 µM TE/mg), whereas ROE demonstrated potent radical scavenging (DPPH (2,2-diphenyl-1-picrylhydrazyl) IC₅₀: 22.81 µg/mL; ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) IC₅₀: 33.6 µg/mL). In vitro scratch assays on human skin fibroblasts (HSF) highlighted ROE and RON as the most effective fractions, reducing wound widths to 0.42 ± 0.04 mm and 0.41 ± 0.005 mm, respectively, within 24 h at 10 µg/mL. These fractions also suppressed LPS(Lipopolysaccharide)-induced nitric oxide production in macrophages by > 70%, underscoring anti-inflammatory synergies. Furthermore, utilising network pharmacology, we identified ten hub target genes associated with wound healing, including IL6 and 1B (Interleukin‑6, -1B), TNF (Tumor Necrosis Factor) and FN1(Fibronectin 1). The findings establish that solvent polarity critically influences bioactive compound recovery, with semi-polar fractions (ROE, RON) optimally balancing antioxidant, anti-inflammatory, and fibroblast-migratory properties for wound healing applications. As a conclusion, R. officinalis is a great natural candidate for valuable bioactive components with promising anti-inflammatory, wound healing, and antioxidant properties. Further phytochemical studies should be performed to isolate the responsible compounds and investigate their mechanism of action.