Multifunctional Hyaluronic Acid-functionalized Liposomes for Targeted Astaxanthin Delivery in Hepatocellular Carcinoma Cells

Introduction: Astaxanthin (AST), a marine-derived carotenoid with potent antioxidant and anticancer activities, is limited in pharmaceutical application by poor aqueous solubility and low bioavailability. In this study, we developed a multifunctional liposomal delivery system integrating Hyaluronic Acid (HA)-mediated targeting, PEGylation, and phytosterol ester-based membrane stabilization for targeted AST delivery in Hepatocellular Carcinoma (HCC) cells. Methods: HA-functionalized AST Liposomes (HA@AST-Lip) with different HA modification ratios were prepared and characterized for physicochemical properties, stability, release, and bioaccessibility. Cellular uptake was evaluated in the Huh-7 HCC cell line, with HA blocking to further investigate HA-CD44-mediated interactions. The in vitro effects on cell viability, migration, apoptosis, and related gene expression were evaluated. Results: HA@AST-Lip exhibited favorable nanoscale characteristics, high encapsulation efficiency, and desirable storage stability. In vitro digestion studies demonstrated sustained release behaviors and significantly improved AST bioaccessibility compared with free AST. Cellular uptake analysis showed enhanced internalization of HA@AST-Lip, possibly via CD44-mediated interactions. Compared with free AST, HA@AST-Lip more effectively inhibited cell proliferation and migration and promoted apoptosis. In addition, HA@AST-Lip modulated the expression of apoptosis- and migration- related genes, which may partially involve NF-κB-related signaling. Discussion: The findings suggest that HA@AST-Lip improved the bioaccessibility and cellular delivery of AST, with enhanced anticancer effects potentially associated with HA-CD44-mediated uptake and NF-κB-related signaling. However, further in vivo studies are needed to validate its therapeutic efficacy and biosafety. Conclusion: These findings demonstrate that HA-functionalized liposomes can improve the delivery performance and anticancer potential of AST, providing a promising lipid-based nanoplatform for HCC-targeted therapy.