Development of liposomal nanocarriers for granulosa cell interaction: Integrating physicochemical characterization...

The development of liposomal formulations tailored for interaction with granulosa cells holds significant promise for applications in reproductive biotechnology. In this study, both anionic and cationic liposomes were formulated using purified and nonpurified phosphatidylcholines and produced using probe sonication followed by extrusion. Liposomes were characterized for size, polydispersity index (PDI), and surface charge, and evaluated for morphology, cytotoxicity, internalization efficiency, and effects on gene expression. Dynamic light scattering revealed nanometric sizes for both formulations (122.1 ± 2.1 nm for anionic and 119.4 ± 2.7 nm for cationic, respectively), with low PDI values (< 0.2), and stable zeta potentials (-31.7 ± 1.4 mV and 48.3 ± 2.6 mV, respectively). Morphology analyses confirmed spherical/near-spherical morphology. Cytotoxicity assays demonstrated that anionic liposomes were biocompatible across all tested concentrations, whereas cationic liposomes induced a dose-dependent effect. Fluorescence microscopy revealed efficient uptake of both liposomal types by granulosa cells, with cationic liposomes exhibiting intracellular fluorescence up to 12 h. Gene expression analysis targeting lipid metabolism, oxidative stress response, steroidogenesis, and apoptosis regulation genes indicated that neither liposomal formulation induced detrimental shifts in cellular homeostasis in the analyzed period. In conclusion, the findings support the use of differently charged liposomal formulations as promising platforms for intracellular delivery in cell-based applications, offering potential for future strategies involving the delivery of genetic material and subsequent modulation of cellular functions.