Laser-programmable glycosaminoglycan-based nanocarriers co-deliver hypocrellin B and doxorubicin for spatiotemporal chemo-photothermal therapy of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) remains lethal due to systemic chemotherapy is halted by cardiotoxicity, multidrug resistance, and poor tumor penetration. To address these limitations, we developed a novel modified chondroitin sulfate/cystamine nanoparticle, termed CS-DHB NPs, that co-delivers doxorubicin (DOX) and the photosensitizer hypocrellin B (HB) and is encapsulated within a chondroitin sulfate/cystamine nanocarrier. These engineered nanoparticles (∼154 nm) demonstrated high drug-loading efficiencies (DOX: 80%; HB: 60%), exceptional serum stability (>72 h), and dual-drug release in laser- and pH-dependent environments. Upon 660 nm laser irradiation (1.0 W cm⁻²), the CS-DHB NPs produced substantial photothermal, photodynamic effects (¹O₂ quantum yield: 0.56), and excellent photostability. The prepared CS-DHB NPs were biocompatible and showed efficient in vitro cytotoxic effects via CCK-8 assays and deep penetration and inhibition of multicellular tumor spheroids (MTS). Ex vivo cellular studies revealed that the targeted delivery mechanism, clathrin-dependent endocytosis, with laser-triggered uptake, enhanced intracellular DOX accumulation twofold. In Hepa1-6 cells, CS-DHB NPs treatment induced pronounced apoptosis (∼51%). In vivo, five intravenous administrations combined with localized laser irradiation resulted in significant tumor regression, 100% survival, minimal systemic toxicity, and no histopathological damage in a Hepa1-6-bearing C57BL/6 mice model. By integrating chemotherapy, phototherapy, and targeted release within a tumor-targeted nanosystem, CS-DHB NPs offer a promising strategy to overcome chemoresistance while preserving healthy tissues, advancing an effective, and clinically translatable approach for synergistic HCC therapy.