The Generation of the Reactive Oxygen Species (ROS)

The ROS generation was assessed by the DPBF method. With blank DPBF as control, the free TCPP, TCPP-MTX-Ag-NP and TCPP-MTX-NP were analyzed (n = 6). In details, acetonitrile was used as the solvent. The final concentration of DPBF in the system was 100 μM and, the nano-carriers according to TCPP and Ag were 10 and 1.5 μg/mL, respectively. The samples in 96-well plate were exposed to 620-nm laser (100 mW/cm²) for 0, 10, 20, 30, and 40 s. For comparison, the same samples were also subjected to darkness. At each time point, the absorbance at 410 nm was measured to see the absorbance decrease caused by the ROS generation (Epoch, Biotek, U.S.A.).