Exosome-niosome hybrid oxygen carrier for protection against acetaminophen-induced acute liver injury

Acetaminophen (APAP) overdose is one of the contributing factors for acute liver injury (ALI), caused by a hypoxic microenvironment, elevated oxidative stress, and pro-inflammatory signaling. In this work, we present an exosome-niosome hybrid oxygen carrier (ENh-OC) composed of polysorbate 80, Pluronic® F-127, medium chain triglyceride oil, and blueberry-derived exosomes. We propose that our delivery system shields the liver tissue from localized hypoxia by maintaining sustained oxygen delivery, and the antioxidant and anti-inflammatory components present in the exosomes enhance the hepatoprotective effect. The hydrodynamic diameter of ENh-OCs is 74.0 ± 30.8 nm with a zeta potential of -13.2 ± 0.5 mV, and oxygen holding capacity of 54.3 ± 1.5 mg L^-1. They have a shelf life of up to 6 months at 4 °C, and an extended oxygen release profile of up to 16 hours under physiological hypoxic conditions. ENh-OCs have demonstrated excellent mitigative effects against hypoxia, reactive oxygen species, and superoxide-induced damage in LX-2 and HepG2 cell lines under 24 hours of sustained hypoxia. Additionally, ENh-OCs have shown a significant protective effect against APAP-induced cytotoxicity in LX-2spheroids when treated after 24 hours of the initial injury phase. The safety profile of ENh-OCs has been established, and efficacy in protection from APAP-induced ALI has been demonstrated in an in vivo murine model. RT-qPCR results verified downregulation of genes related to hypoxia (HIF-1α, VEGF-A), oxidative stress (Nrf2, HO-1), and inflammation (IL-1β, TNF-α) both in vitro and in vivo. Furthermore, histological analysis revealed protective effects of ENh-OCs against centrilobular necrosis and excessive immune-infiltration. We propose that the novel oxygen nanocarrier platform introduced here might act as a protective agent against APAP-induced ALI.