o:38808 Development and Characterization of Natamycin-Loaded Liposomes for Potential Topical Application: Influence of Preparation Method and Phospholipid Composition en Background/Objectives: Natamycin is an effective antifungal limited by poor solubility. This study aimed to develop and characterize natamycin-loaded liposomal vesicles as a biocompatible delivery system to improve stability and achieve controlled release for potential topical application in the treatment of fungal infections. Methods: Formulations were prepared using two phospholipid mixtures (Lipoid S100 and Phospholipon 90H) via standard (thin-film) and proliposome methods. Evaluation included encapsulation efficiency (EE%), particle size, zeta potential, the polydispersity index (PDI), and rheological properties. In vitro release kinetics were compared to a natamycin solution. Antifungal efficacy was tested against four Candida strains to determine minimum inhibitory and fungicidal concentrations (MICs and MFCs, respectively) and biofilm inhibition, while biocompatibility was assessed via keratinocyte viability assays. Results: Formulations achieved high encapsulation (~90%). Natamycin incorporation improved homogeneity and reduced particle diameters, particularly in proliposome-derived vesicles, suggesting strong drug–lipid interactions. Preparation method and lipid type significantly influenced properties; thin-film formulations showed a lower PDI and higher stability. Diffusion was twofold slower than the control, with Lipoid S100 proliposomes providing the most sustained release. The liposomes demonstrated robust antifungal activity (MICs: 0.00625–0.2 mg/mL) and effective biofilm inhibition against C. krusei. While high concentrations moderately reduced keratinocyte viability, lower doses remained biocompatible and slightly stimulatory. Conclusions: Lipid composition and preparation methods have minimal impact on the physical properties and in vitro release profiles of natamycin liposomes. These vesicles provide a dose-dependent, biocompatible platform for the controlled delivery of antifungals, showing significant in vitro inhibitory activity against Candida growth and biofilm formation. skin application; controlled release; nanoparticle tracking analysis; antibiofilm activity; natamycin; liposomal vesicles 1552099 https://doi.org/10.3390/ph19050710 2026-05-04T10:12:26.833Z 44 yes 46 Natalija Čutović Institute for Medicinal Plants Research “Dr Josif Pančić”, Belgrade, Serbia Petar Batinić Institute for Medicinal Plants Research “Dr Josif Pančić”, Belgrade, Serbia person Tatjana Marković Institute for Medicinal Plants Research “Dr Josif Pančić”, Belgrade, Serbia person Andrea Pirković University of Belgrade, Institute for the Application of Nuclear Energy INEP, Serbia doktor bioloških nauka person 0000-0002-1182-4895 Ninoslav Mitić University of Belgrade, Institute for the Application of Nuclear Energy INEP, Serbia doktor bioloških nauka person 0000-0003-2756-6517 Jovana Petrović Department of Plant Physiology, Institute for Biological Research “Siniša Stanković”—National Institute of Republic of Serbia, University of Belgrade, Belgrade, Serbia person Aleksandra A. Jovanović University of Belgrade, Institute for the Application of Nuclear Energy INEP, Serbia doktor tehnoloških nauka person 0000-0001-5394-0125 application/pdf 6111109 https://phaidrabg.bg.ac.rs/o:38808 no yes 16 70 92000001 11A29 11A21 11A2107 Development and Characterization of Natamycin-Loaded Liposomes for Potential Topical Application: Influence of Preparation Method and Phospholipid Composition 710 19 5 Pharmaceuticals Basel, Switzerland MDPI 2026