• • SSD-CBD nanoparticles were prepared at a 3:1 mass ratio (SSD:CBD) using nano co-precipitation, achieving a synergistic effect against HepG2 cells with a combination index (CI) of 0.79, indicating moderate synergy. This provides a quantitative baseline for co-delivery ratio optimization in clinical translation.
• • The nanoparticles exhibit pH-responsive drug release: specific release in tumor microenvironment (pH 6.8) while remaining stable at physiological pH 7.4. This differential release profile is critical for minimizing off-target toxicity and enhancing tumor-selective payload delivery.
• • Mechanistic studies via MTT, Annexin V-FITC/PI flow cytometry, and caspase activity assays confirm that SSD-CBD nanoparticles induce HepG2 cell death primarily through the mitochondrial apoptotic pathway, offering a defined cell death mechanism for regulatory submission.
• • The carrier-free self-assembly strategy simultaneously improves CBD's delivery efficiency and enables precise synergistic drug co-administration, bypassing the need for synthetic excipients. However, in vivo pharmacokinetics, tissue distribution, and potential SSD hepatotoxicity in nanoformulation remain uncharacterized, representing a critical gap for further development.