Antimicrobial resistance has intensified the search for novel antimicrobial and wound-healing agents. Ludwigia octovalvis (Jacq.) P.H. Raven is traditionally used to treat infections, inflammation, and skin disorders, although its in vitro biotechnological potential remains largely unexplored. This study reports the first successful establishment of L. octovalvis callus cultures, their characterization by gas chromatography-mass spectrometry (GC-MS), and the evaluation of their antimicrobial and wound-healing activities. Friable calluses were induced from leaf and node explants of axenic seedlings using combinations of 6-benzylaminopurine (BAP), kinetin (KIN), 2,4-dichlorophenoxyacetic acid (2,4-D), and α-naphthaleneacetic acid (NAA). Optimal callus induction was achieved in leaf explants cultured on full-strength Murashige and Skoog (MS) medium supplemented with BAP (4.44 µM) and 2,4-D (0.45 µM). After six months of subculture, the calluses showed morphological uniformity and stable biomass production. Growth kinetics followed a specific growth rate of 0.047 d⁻¹ and a doubling time of 14.54 days. GC-MS analysis identified fatty acids and phytosterols as the main components. The ethyl acetate extract significantly improved wound healing in vivo, while the methanolic extract exhibited antibacterial activity against Staphylococcus aureus and methicillin-resistant S. aureus (MRSA). These findings demonstrate that L. octovalvis callus cultures constitute a sustainable source of bioactive compounds with promising therapeutic potential.

Due to Ludwigia octovalvis’ aquatic habitat’s vulnerability to climate change, this study developed an in vitro regeneration system using indirect organogenesis to ensure sustainable production of biomass and secondary metabolites. Treatment T16 (0.1 mg/L BAP and 1.0 mg/L NAA) was identified as the optimal hormonal regimen for callus induction and shoot differentiation. Phytochemical analysis by GC-MS revealed that seedlings regenerated under treatment T16 exhibited a diverse profile of 18 phytoconstituents, enhancing the accumulation of phytosterols, terpenes, and tocopherols. In vitro biological evaluation demonstrated that T16 extract possesses significant antibacterial activity (MIC < 62.5 µg/mL) against methicillin-resistant Staphylococcus aureus, and moderate antioxidant capacity. T16 extract showed anti-inflammatory effects superior to indomethacin at a low quantity (0.5 mg/ear) in adult CD1 mice of both sexes. In conclusion, the indirect organogenesis of L. octovalvis not only conserves the species but also optimizes its pharmacological potential, consolidating it as an efficient biotechnological platform for the development of advanced phytopharmaceuticals.