Multifunctional effects of Zizyphus mauritiana leaf extract on oral pathogens and wound healing: an integrative in vitro and in vivo study

Post-extraction infections and delayed oral wound healing remain clinical challenges due to bacterial biofilms, oxidative stress, and inflammation. Natural plant-derived compounds offer promising alternatives to conventional antimicrobials. Zizyphus mauritiana (daun bidara), widely used in traditional Indonesian medicine, has not been comprehensively evaluated for its potential role in oral infection control and wound repair. This study investigated the antibacterial, antibiofilm, antioxidant, and anti-inflammatory activities of ethanolic extract of Z. mauritiana (ZM) leaves using both in vitro and in vivo models. The antibacterial activity of ZM extract was tested against Prevotella intermedia, Fusobacterium nucleatum, Streptococcus sanguinis, and Treponema denticola using the microdilution method to determine minimum inhibitory (MIC) and bactericidal (MBC) concentrations. Antibiofilm activity was assessed using a 96-well plate biofilm assay and confirmed by quantitative polymerase chain reaction (qPCR). Antioxidant activity was evaluated using the DPPH assay to determine the IC₅₀ value. For in vivo evaluation, gels containing 50% and 100% ZM extract were applied to tooth extraction sockets in Sprague-Dawley rats (n = 24). On day 3, wounds were analyzed for tumor necrosis factor-α (TNF-α) levels by ELISA and for histological inflammation using immunohistochemistry (IHC). ZM extract exhibited strong antibacterial activity (MIC = 125-500 mg/mL; MBC = 250-500 mg/mL) and significantly inhibited biofilm formation (p < 0.05). The extract showed moderate antioxidant activity (IC₅₀ = 0.13703 mg/mL). In vivo, the 50% ZM gel markedly reduced TNF-α levels (118.7 pg/mL) and produced sparse IHC staining, indicating reduced inflammation and improved healing. Ethanolic extract of Zizyphus mauritiana leaves demonstrates multifunctional bioactivities and holds promise as a natural therapeutic agent for managing oral infections and enhancing post-extraction wound healing.