Targeting Behavior and Cellular Uptake Mechanisms of Curcumin in Folate-Transferrin Dual-Targeted Liposome Systems

Targeted nanoliposomes constitute a critical platform for improving therapeutic outcomes in oncology. However, their cellular recognition and uptake efficiency are frequently compromised by the heterogeneity of receptor expression on tumor cells and the inherent limitations of single-ligand targeting strategies. To overcome these challenges, we developed a dual-targeted liposomes system comodified with folic acid and transferrin (FA/Tf-F127-Lps) for curcumin delivery, leveraging dual-receptor synergy to enhance targeting and cellular internalization. The FA/Tf-F127 conjugate was synthesized via chemical coupling and used to prepare curcumin-loaded liposomes through thin-film hydration combined with dynamic high-pressure microfluidics, achieving a drug encapsulation efficiency of 86.69%. Cellular experiments demonstrated that the dual-targeted system exhibited the strongest cytotoxicity against C6 glioma cells, with IC50 values of 1.297 mg/L at 48 h and 1.175 mg/L at 72 h, and its cellular uptake was significantly higher than that of single-targeted and nontargeted controls. Uptake mechanistic studies revealed that internalization was energy-dependent and involved clathrin-mediated, caveolae-mediated, and macropinocytic pathways, and could be competitively inhibited by free folic acid and transferrin. This dual-targeted system shows substantial promise for treating receptor-overexpressing tumors such as glioma by achieving enhanced specificity and intracellular delivery. The strategy provides a feasible experimental basis for addressing receptor heterogeneity and improving targeting precision in glioma therapy, while also offering an approach for developing next-generation personalized nanomedicines.