P. lanceolata L. and P. major L. extracts in 3D-printed polysaccharide dressings for improved wound healing in vitro

Plantago species have long been used in traditional medicine for wound healing, yet systematic validation remains limited. To explore their potential, we extracted active compounds from Plantago major (PM) and Plantago lanceolata (PL) using various solvents and incorporated them into a novel wound care formulation. The PM extract contained primarily plantamajoside, followed by verbascoside and aucubin, while the PL extract was rich in verbascoside, aucubin, and catalpol. Extract safety was evaluated on skin fibroblasts and keratinocytes using an MTT assay. PM extract (1000.0 μg/mL) increased fibroblast metabolic activity to 1.2 without cytotoxicity toward keratinocytes, while PL extract (100.0 μg/mL) enhanced keratinocyte viability to 1.3 without significant fibroblast effects. Methanol and ethanol extracts of PM and PL were incorporated into a polysaccharide-based wound dressing (WD) composed of methylcellulose, alginate, and nanofibrillated cellulose. The WDs were comprehensively characterized for swelling, degradation, and active compound release, as well as structural features using scanning electron microscopy, nanotomography, and atomic force microscopy. Mechanical strength and adhesion testing confirmed suitable handling and application properties, while anti-inflammatory assays demonstrated additional therapeutic potential of the extracts. Biocompatibility testing showed that the incorporation of 0.5 wt% extract increased fibroblast viability to 1.39 for PM WD and 1.5 for PL WD. Scratch assays on fibroblasts demonstrated enhanced wound closure, with PM- and PL-infused WDs closing wounds by 64% ± 7% and 52% ± 7%, respectively, compared to 29% ± 6% for the control and 33% ± 11% for the WD without extract. These results highlight the multifunctional potential of PM and PL extracts in wound healing applications. • Plantago species have been used in traditional medicine for wound healing, but scientific validation is still required to confirm their effectiveness. P. major (PM) and P. lanceolata (PL) extracts were prepared and quantified. • Active compounds in extracts – PM extract contained plantamajoside (PMJ), verbascoside (VER), and aucubin (AUC), while PL extract was rich in VER, AUC, and catalpol (CAT), highlighting their bioactive potential. • Cellular effects & safety – PM extract (1000.0 μg/mL) boosted fibroblast metabolic activity (1.2×) with no keratinocyte toxicity, while PL extract (100.0 μg/mL) increased keratinocyte viability (1.3×). • Wound dressing formulation – PM and PL extracts were incorporated into a methylcellulose (MC), alginate (ALG), and nanofibrillated cellulose (NFC)-based wound dressing for controlled release and wound healing. • Improved wound healing – Scratch assays showed PM- and PL-infused wound dressings improved wound closure (64 ± 7% and 52 ± 7%) compared to the control (29 ± 6%), confirming their healing potential.