In vitro and in vivo evaluation of liposomes constructed with Rg3 as a cholesterol substitute for tumor therapy

Conventional cholesterol liposomes have poor stability, rapid clearance, and toxicity. This study used Rg3 to replace cholesterol for constructing curcumin-loaded liposomes (PCRL) and evaluated their potential for enhanced hepatocellular carcinoma therapy. Curcumin-loaded liposomes were prepared with Rg3 (PCRL) and cholesterol (PCCL) via thin-film hydration. Physicochemical properties, protein corona, in vitro behavior, in vivo tumor targeting, antitumor efficacy, and biosafety were assessed in H22 tumor-bearing mice. PCRL exhibited smaller particle size, superior storage/plasma stability, and comparable membrane packing to PCCL. Protein corona analysis showed reduced adsorption of immunoglobulin-like and complement proteins, prolonging blood circulation > 24 h and enhancing tumor accumulation. In vitro, PCRL showed good biocompatibility, reduced curcumin cytotoxicity, enhanced cellular uptake, and the slowest drug release. In vivo imaging demonstrated stronger and more persistent tumor fluorescence with lower liver/spleen retention. PCRL significantly inhibited tumor growth. Safety assessment revealed low hemolysis, no hepatorenal toxicity, normal spleen index, and good body weight tolerance. Curcumin-loaded liposomes with Rg3 substituting cholesterol maintain membrane integrity while prolonging circulation, improving tumor targeting/efficacy, and enhancing biosafety, offering a promising strategy for next-generation stealth liposomal delivery systems.