Biological Activities of Stachys rupestris, Development of S. rupestris Extract-Loaded Alginate Films as Wound Dressing

<b>Background/Objectives</b>: Regardless of the underlying cause, wound infections are among the most common complications associated with wound formation. The increasing prevalence of antibiotic resistance poses significant challenges in wound management. Due to their favorable therapeutic properties, alginate films have recently emerged as promising biomaterials for wound treatment. <b>Methods</b>: The petroleum ether, chloroform, and methanol extracts of the endemic plant <i>Stachys rupestris</i> were prepared using the maceration technique. The antimicrobial activity of the extracts and the extract-loaded alginate film was evaluated by agar well diffusion and microdilution assays, while their antibiofilm activity was assessed by crystal violet staining in microplates. The anti-infective potential was investigated using the <i>Caenorhabditis elegans</i> infection model, the phytochemical composition was analyzed by HPLC-DAD, and cytotoxicity was determined by the MTT assay. The alginate film was prepared by the solvent casting method and characterized using FTIR spectroscopy and light microscopy. <b>Results</b>: All extracts demonstrated antimicrobial activity, with the methanol extract exhibiting the most potent antimicrobial and antibiofilm effects. Quinic acid was identified as the major constituent. Both the methanol extract and the film displayed no cytotoxic effects and showed significant antimicrobial and antibiofilm activities. <b>Conclusions</b>: The <i>S. rupestris</i> methanol extract-loaded film exhibited strong antimicrobial and antibiofilm properties, indicating its potential as a valuable therapeutic agent in supporting wound healing.