Ex vivo hemocompatibility and hemorheological evaluation of liposomal nanocarriers containing furosemide and blueberry extract

Abstract

Bovine mammary edema is a frequent clinical condition that impairs animal welfare and causes economic losses in livestock production, motivating the search for improved therapeutic strategies. In this work, liposomal nanocarriers were developed to incorporate Furosemide (Fr), a loop diuretic, and an anthocyanin-rich blueberry extract (BE), a natural antioxidant source. The antioxidant capacity of BE was confirmed by the DPPH assay, yielding an IC₅₀ of 27 mg/mL, which was used to define the concentration employed for the liposomal formulation. The selected formulation exhibited a hydrodynamic diameter of 278.5±9.2 nm, a polydispersity index of 0.28±0.06, and a negative zeta potential, indicating the formation of nanoscale vesicles. Encapsulation efficiency analysis revealed moderate loading in single-compound systems (Fr: 51.12%; BE: 74.93%). However, in the co-loaded formulation, a marked decrease in encapsulation efficiency was observed (Fr: 0.8%; BE: 28.02%), indicating limited incorporation of Furosemide under the studied conditions. FT-IR analysis suggested interactions between the liposomal matrix and the incorporated compounds while preserving their main functional groups. However, BE-loaded liposomes did not exhibit detectable antioxidant activity under the tested conditions, likely due to limited accessibility of the encapsulated compounds to the DPPH radical and reduced effective concentrations in the liposomal formulations. Hemocompatibility studies using bovine red blood cells showed negligible hemolysis (<5%) and no significant alterations compared to the negative control. Additionally, hemorheological evaluation indicated that the formulations did not adversely affect erythrocyte behavior. Overall, these results demonstrate that while liposomal systems provide a suitable platform for incorporating BE and evaluating physicochemical and hemocompatibility properties, the coloading with Furosemide is markedly limited under the present conditions. These findings highlight the need to optimize formulation parameters further to achieve effective co-encapsulation in future studies.