Development of engineered magnetic liposome/exosome hybrid as a novel caffeine nanocarrier for restraining liver fibrosis induced in rats

Liver fibrosis, a severe health issue linked to liver injury, inflammation, and cell death, often progresses to liver cancer without effective treatments. In this study, an exosome-nanoliposome hybrid loaded with caffeine (Caff) was developed for the management of liver fibrosis. The role of superparamagnetic iron oxide nanoparticles (SPION) in promoting targeted delivery to the liver was also investigated. Caff-loaded liposomes with variable phospholipid and stearylamine molar ratios were prepared and characterized. The formula with optimized characteristics (Caff/liposomes) selected by Design Expert exhibited a particle size of 179.73 ± 3.1 nm, a polydispersity index of 0.19 ± 0.02, an entrapment efficiency of 89.79 ± 21%, and a zeta potential of -35.50 ± 1.1 mV, and was used to encapsulate SPION. Caff/SPION liposomes were further loaded into exosomes, isolated from bone marrow-mesenchymal stem cells, and visualized by TEM. The biological experiment was conducted on rats with thioacetamide-induced liver injury to reveal the efficacy of exosome-Caff/SPION liposome hybrid (F1) in comparison to exosome-Caff/liposomes (F2) and exosome-SPION/liposomes (F3). Among all the tested formulations, F1 demonstrated the most promising results regarding liver function markers (ALT and AST), inflammatory molecules (IL-6, IL-10, TLR-4, Myd88, NF-κB), fibrotic protein (α-SMA), hepatocyte proliferative capacity (PCNA), as well as liver histoarchitecture. This finding suggests that the engineered Caff/SPION liposome-exosome hybrid can effectively enhance the targeting of the drug and the homing of exosomes to the injured liver, thereby representing a novel strategy for treating liver fibrosis.