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. 2021 Jun 30;8:409–419. doi: 10.1016/j.bioactmat.2021.06.021

Fig. 3.

Fig. 3

In vitro mild PTT-enhanced SDT of HH-Ti3C2-PEG NSs. (a) Schematic illustrating of HH-Ti3C2-PEG NSs for the mild PTT-enhanced SDT. (b) Relative cell viability of 4T1 cells with the HH-Ti3C2-PEG NSs with various concentrations for 6, 12, and 24 h (n = 6 biologically independent samples). (c) Relative cell viability of 4T1 cells after different treatments (G 1-G 7, detailed at the end of the caption) (***P < 0.001, **P < 0.01, or *P < 0.05). (d) Confocal images of 4T1 cells incubated with Cy5.5-conjugated HH-Ti3C2-PEG NSs for different times with/without 1064 nm laser irradiation. (e) Quantitative analysis of the fluorescence intensity in (d) (n = 6 cells examined over independent micrographs). (f) Flow-cytometry apoptosis assay of 4T1 cells after different treatments stained with Annexin-FITC and PI. (g) Confocal images of 4T1 cells after incubation HH-Ti3C2-PEG NSs followed by staining with Calcein AM (green, live cells) and propidium iodide (red, dead cells) after different treatments (G 1-G 7). (h) Confocal images of 4T1 cells stained with DAPI (blue, nuclei) and DCFH-DA (green, intracell ROS) after various treatments (G 1-G 7). G 1: Control, G 2: HH-Ti3C2-PEG, G 3: Laser, G 4: US, G 5: HH-Ti3C2-PEG + Laser, G 6: HH-Ti3C2-PEG + US, G 7: HH-Ti3C2-PEG + Laser + US. HH-Ti3C2–PEG NSs: 50 ppm, NIR laser: 1064 nm, < 1W ▪ cm−2, 10 min, T < 42 °C; US irradiation: 40 kHz, 3W ▪ cm−2, 1 min per cycle, 5 cycles. Error bars = standard deviation (n = 6). Data are presented as mean values ± SD. A representative image of three biological replicates from each group is shown.