Improved Method for Solid Lipid Nanoparticle Preparation Based on Hot Microemulsions: Preparation, Characterization, Cytotoxicity, and Hemocompatibility Evaluation

Abstract

The ease of application and no requirement of extra energy input make the microemulsion method favorable for solid lipid nanoparticles (SLNs) production. Very limited data are available to date on preparation of SLNs from pre-screened microemulsion phase diagrams. The purpose of this study was to investigate the microemulsion formation area with solid lipids using hot ternary phase diagrams at elevated temperatures and to use selected microemulsions for SLN production. Also, we aimed to characterize obtained SLNs in terms of physicochemical properties, in vitro cell toxicity, and hemolysis. Phase diagrams of solid lipids were screened at elevated temperatures and oil-in-water microemulsion regions were determined. Microemulsions were selected, and SLNs were produced by modification of the microemulsion dilution method and characterized in terms of visual appearance, turbidity, particle size, and zeta potential. Cytotoxicity of nanoparticles was tested on L929 mouse skin fibroblast cells. Hemolytic potential was assessed in vitro using freshly isolated erythrocytes. The phase diagram screening and the modified hot microemulsion dilution method enabled production of SLNs with particle size below 100 nm. We found evidence that the solid lipids in the SLNs produced by the new method remain in supercooled liquid state. Nanoparticles prepared by the new method exhibit lower toxicity on L929 cells and have lower hemolytic potential than the formulations prepared by direct mixing of the components. The method can be used to prepare SLNs with controllable composition and small particle size below 100 nm. These SLNs are low toxic and can be used for drug delivery purposes.

Keywords: microemulsion method; solid lipid nanoparticles; supercooled liquid state; ternary phase diagram.