Comparative Phytochemical Profiling and Wound Healing Potential of Scabiosa pseudograminifolia Hub.‐Mor. and Scabiosa hololeuca Bornm.: UHPLC ‐ HRMS / MS Analysis and Fibroblast‐Based Evaluation

ABSTRACT Species of the genus Scabiosa have long been used in traditional medicine for the treatment of skin disorders and inflammatory conditions; however, comparative information on their chemical composition and biological activity remains limited. In this study, the phenolic composition and functional properties of Scabiosa pseudograminifolia Hub.‐Mor. and Scabiosa hololeuca Bornm. were investigated using microwave‐assisted extraction combined with UHPLC‐HRMS/MS analysis and in vitro biological assays. Methanolic and aqueous extracts were analyzed for their phenolic constituents and evaluated in NIH/3T3 fibroblasts for intracellular reactive oxygen species (ROS) modulation, cell viability, and wound healing activity. Antimicrobial activity was additionally assessed using a broth microdilution method against selected bacterial and fungal strains. All extracts significantly reduced tert‐butyl hydroperoxide (tBHP)‐induced intracellular ROS levels, demonstrating antioxidant activity in the cellular model. Extracts of S. pseudograminifolia showed a pronounced wound healing effect, promoting more than 95% wound closure after 48 h at low concentrations, whereas those of S. hololeuca exhibited more moderate activity. The concentrations responsible for these effects were confirmed to be non‐cytotoxic. Distinct species‐ and solvent‐dependent differences in phenolic composition were observed. In S. pseudograminifolia , chlorogenic acid was identified as the major compound in both methanolic and aqueous extracts, while the aqueous extract also contained high levels of 4‐ O ‐caffeoylquinic acid. In contrast, S. hololeuca extracts contained lower amounts of chlorogenic acid and were characterized by relatively higher levels of caffeic and protocatechuic acids. Antimicrobial activity was weak to moderate, suggesting that the principal biological relevance of these extracts is associated with oxidative stress modulation and wound repair rather than direct antimicrobial effects. This study represents the first integrated phytochemical and biological evaluation of these two closely related taxa.