Chitosan loaded copper and selenium nanoparticles synergistically inhibit HepG2-hepatic cancer cells viability and...

Chitosan-stabilized copper-selenium nanoparticles (CS-Se-Cu NPs) were synthesized via a facile green reduction method using ascorbic acid as the reducing agent. The nanoparticles were comprehensively characterized by powder X-ray diffraction (PXRD), Fourier-transform infrared spectroscopy (FTIR), field emission scanning electron microscopy (FESEM), and energy-dispersive X-ray spectroscopy (EDX). PXRD confirmed a pure monoclinic crystalline phase (PDF 00-049-1456) with a crystallite size of approximately 18 nm and minimal lattice strain (ε = -0.002). FTIR indicated chitosan capping through characteristic amide and hydroxyl peaks, while FESEM revealed spherical to semi-spherical morphology (approximately 50-100 nm), and EDX verified a Cu: Se atomic ratio of about 1:1. MTT assay demonstrated potent, concentration- and time-dependent cytotoxicity against HepG2 liver cancer cells (IC₅₀, 24 h: 1.8 ± 0.2 mg/mL; 72 h: 0.9 ± 0.1 mg/mL), with fourfold higher selectivity over normal NIH-3T3 fibroblasts (IC₅₀, 72 h: >5 mg/mL). A biphasic dose-response was observed in HepG2 cells, with viability above 80% at low doses (≤ 0.63 mg/mL, 24 h) decreasing to below 30% at high doses (≥ 1.25 mg/mL, 72 h). In the chicken chorioallantoic membrane (CAM) assay, CS-Se-Cu NPs exerted dose-dependent anti-angiogenic effects: at 0.8 mg/mL, vessel number decreased by 62% (24.5 ± 2.1 to 9.2 ± 1.3; p < 0.001) and total length by 63% (28.5 ± 1.8 to 10.7 ± 1.2 mm; p < 0.001), accompanied by 32% and 33% reductions in embryonic length and weight, respectively. These results underscore the synergistic potential of CS-Se-Cu NPs as dual cytotoxic and anti-angiogenic agents for liver cancer therapy.