2.10. Dual-Ligand Targeted Cytotoxicity

In a typical procedure for evaluating the NIR light-triggered cytotoxicity of NC1 + NC2 mixture, HeLa cells or A549 cells cultured in a 96-well plates were treated with NC1 with gradient concentrations and NC2 with a fixed concentration (300 μg/mL) for 6 h at 37 °C, followed by 980 nm light (1 W/cm²) irradiation for 10 min. After being incubated for another 42 h, the viabilities of treated cells were investigated via an MTT assay. To evaluate the influence of GSH, HeLa cells were treated with 0.1 mM BSO overnight before adding the NC1 + NC2 mixture [30]. To analyze the cytotoxicity of single nanocarrier, NC1 or NC2 with gradient concentrations were incubated with HeLa cells seeded in 96-well plates for 48 h with/without 980 nm light irradiation for 10 min. To evaluate the toxicity of blank nanocarriers and NIR light irradiation, gradient concentrations of non-DOX-loaded NC1 or NC2 were incubated with HUVECs or HeLa cells for 48 h with or without 980 nm light irradiation (1 W/cm²) for 10 min. After incubation, the cell viabilities were investigated via an MTT assay.