Fig 1.
Effect of PDT on cell viability in ICG-lactosome-treated HuH-7 cells.
Cells were divided into four groups: control, ICG-lactosome, laser, and PDT (ICG-lactosome + laser). The laser and PDT groups were irradiated with (A) 18 J/cm2 (190 mW/cm2 and 95 s), (B) 18 J/cm2 (340 mW/cm2 and 55 s), (C) 100 J/cm2 (190 mW/cm2 and 525 s), and (D) 100 J/cm2 (340 mW/cm2 and 300 s). Cell viability after irradiation was measured by the MTT assay at the indicated times (n = 4 dishes/time/group). *P<0.001 ICG-lactosome vs. other groups. OD570, optical density at 570 nm.
Fig 2.
Effect of PDT on morphological changes in ICG-lactosome-treated HuH-7 cells.
Morphological changes were observed with a phase-contrast microscope at 96 h after each treatment. (A) Control group. (B) ICG-lactosome group. (C) laser (100 J/cm2 (340 mW/cm2 and 300 s)) group. (D) PDT (ICG-lactosome + laser) group. Representative images from four experiments are shown.
Fig 3.
Effect of laser irradiation on temperature in ICG-lactosome-treated HuH-7 cells.
The culture medium temperature was measured using a thermo-camera in the laser and PDT groups with low or high laser irradiation. (A) 18 J/cm2: low laser (190 mW/cm2 and 95 s), high laser (340 mW/cm2 and 55 s), low laser PDT, and high laser PDT (n = 4 dishes/group). (B) 100 J/cm2: low laser (190 mW/cm2 and 525 s), high laser (340 mW/cm2 and 300 s), low laser PDT. and high laser PDT (n = 4 dishes/group).
Fig 4.
In vivo fluorescence imaging in the ICG-treated and ICG-lactosome-treated mice with subcutaneous tumors.
Fluorescence imaging was acquired using an IVIS system in the ICG-treated mice (A and C) and ICG-lactosome-treated mice (B and D). After injection of ICG or ICG-lactosome, the brightness of the tumor (open circles) and non-tumor (contralateral inguinal, closed circles) areas was measured at the indicated times. P<0.001 between the tumor and non-tumor regions in the ICG-lactosome group; n = 5/group.
Fig 5.
Effect of PDT on tumor growth in the ICG-treated and ICG-lactosome-treated mice with subcutaneous tumors.
At 48 h after ICG (open circles) or ICG-lactosome (closed circles) injection in the tumor implanted-mice, laser irradiation (500 mW/cm2 and 200 s, 100 J/cm2) was carried out, and the tumor volumes were measured at the indicated times. P<0.001 between the ICG and ICG-lactosome groups; n = 6/group.
Fig 6.
Effect of laser irradiation on tumor temperature in the ICG-treated and ICG-lactosome-treated mice with subcutaneous tumors.
At 48 h after ICG (yellow) or ICG-lactosome (brown) injection or no injection (control, dark) in the tumor implanted-mice, the laser irradiation (500 mW/cm2 and 200 s, 100 J/cm2) treatment was initiated, and the temperatures were measured at the indicated times. P<0.001 between the ICG and ICG-lactosome groups; n = 6/group.
Fig 7.
Effect of PDT on cells of subcutaneous tumors in the ICG-treated and ICG-lactosome-treated mice.
Tumors were obtained for examination of apoptosis at 24 h after irradiation (500 mW/cm2 and 200 s, 100 J/cm2) in the ICG (A) and ICG-lactosome (B) groups. TUNEL staining was performed on the specimens. (C) Apoptotic indexes in the ICG-treated and ICG-lactosome-treated mice with subcutaneous tumors. P = 0.001 between the ICG and ICG-lactosome groups; n = 5/group.