图1 IR780@Cu9S8的合成及表征

Fig.1 Synthesis and characterization of IR780@Cu9S8 nanoparticles. A: TEM images of Cu9S8 and IR780@Cu9S8 nanoparticles. B: XRD spectra of IR780@Cu9S8. C: UV-Vis spectra of Cu9S8, IR780 and IR780@Cu9S8 nanoparticles. D: Hydrodynamic diameter of Cu9S8 and IR780@Cu9S8 nanoparticles.

图2 IR780@Cu9S8的光热性能评估

Fig.2 Evaluation of the photothermal performance of IR780@Cu9S8. A: Photothermal curves of PBS, Cu9S8, and IR780@Cu9S8 (50 μg/mL) under 808 nm irradiation. B: Heating curves of IR780@Cu9S8 (50 μg/mL) aqueous solution under various laser powers (0.5, 1.0, 1.5, 2.0, 2.5 W/cm²). C: Photothermal curves of IR780@Cu9S8 aqueous suspensions at different concentrations under 808 nm light irradiation (1.5 W/cm²). D: Photothermal stability curve of IR780@Cu9S8 aqueous suspension (50 µg/mL). E, F: Temperature elevation and photothermal conversion efficiency of IR780@Cu9S8 under 808 nm irradiation.

图3 IR780@Cu9S8的体外细胞毒性评估

Fig.3 In vitro cytotoxicity of IR780@Cu9S8 upon 808 nm irradiation. A: Viability of KYSE150 and HUVEC cells after 24 h co-incubation with IR780@Cu9S8. B: Viability of KYSE150 cells with different treatments. C: Viability of KYSE150 cells after 6 h incubation with IR780@Cu9S8 followed by 808 nm irradiation, fitted with a nonlinear dose-response curve (logistic model). D: Fluorescence images of calcein-AM/PI-stained KYSE150 cells after different treatments. E: Representative confocal images of ROS in KYSE150 cells after different treatments. F: Mean fluorescence intensity of ROS in KYSE150 cells. Data are presented as Mean±SD (n=3).

图4 不同处理后KYSE150细胞线粒体膜电位荧光染色

Fig.4 Fluorescence images showing the mitochondrial membrane potential of KYSE150 cells stained with JC-1.

图5 IR780@Cu9S8对体外肿瘤细胞功能的抑制作用评估

Fig.5 Evaluation of the inhibitory effect of IR780@Cu9S8 on malignant behaviors of tumor cells in vitro. A, C: Colony-forming assay of the cells after different treatments. B, D: Changes in cell invasion capacity after the treatments. E, F: Relative migration rate of the cells at 24 h after different treatments in wound-healing assay and representative images of the cells at 0 h and 24 h. Data are presented as Mean±SD (n=3).

图6 IR780@Cu9S8的体内抗瘤疗效评估

Fig.6 Evaluation of the in vivo antitumor efficacy of IR780@Cu9S8. A:Schematic illustration of the strategy for treatment. B:Changes in tumor volume over time in the tumor-bearing mice. C:Photographs of excised tumors. D:Comparison of tumor weights at the end of the treatment among the groups. Data are presented as Mean±SD (n=3).

图7 IR780@Cu9S8的体内抗瘤疗效评估(HE染色、免疫荧光染色)

Fig.7 Evaluation of the in vivo antitumor efficacy of IR780@Cu9S8. A: HE staining, Ki-67 immunohistochemistry and TUNEL fluorescence staining of tumor sections. B: Percentage of Ki-67 positive cells among all cells. C: Percentage of TUNEL positive cells among all cells. Data are presented as Mean±SD (n=3).

图8 IR780@Cu9S8的体内生物安全性检测

Fig.8 Evaluation of in vivo biocompatibility of IR780@Cu9S8. A: Body weights of male C57BL/6 tumor-bearing mice after different treatments. B: Hemolysis of male C57BL/6 mouse blood after treatment with different concentrations of IR780@Cu9S8. Data are presented as Mean±SD (n=3).

图9 小鼠血常规及血生化主要指标

Fig.9 Hematological and biochemical parameters analysis results of tumor-bearing male C57BL/6 mice after different treatments. Data are presented as Mean±SD (n=3).

图10 小鼠重要脏器HE染色

Fig.10 HE staining of various organs from tumor-bearing male C57BL/6 mice after different treatments.