At 24 hours after the final intravenous injection, the major organs (heart, liver, spleen, lungs, and kidney) of the mice were excised, fixed, sectioned, and stained with H&E for histological analysis. The tissue slices were subjected to microscopic observation (Leica DMI 6000B). Before organ collection, an electronic balance was used to determine the body weights of the mice. At 24 hours after the final intravenous injection, blood was sampled from the venous plexus of the mice’s eyes. The blood samples were subjected to immediate centrifugation at 3000g for 5 min at 4°C. Hematological analysis was then performed on the supernatant. The blood biochemical analysis comprised determining the ALT, AST, blood urea nitrogen, and creatinine levels using Modular Analytics (Roche, Germany). ALT and AST were used as indicators of hepatic functions, and blood urea nitrogen and creatinine were used as indicators of renal functions. To assess the immunotoxicity of the various formulations, we determined cytokines present in the serum using ELISA kits for IL-1β, IL-6, IFN-α, and TNF-α (Abcam).

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