The cytotoxicity
of Hu-ink was assessed by CCK-8 assay. Approximately 1 × 104 HCT-116 and SW-620 cells were plated in 96-well plates and
cultured for 24 h at the standard cell culture environment. The cells
were cultured with samples (0–200 μg/mL) for 24 h and
then washed twice with PBS; 100 μL of RPMI 1640 cell medium
and 10 μL of CCK-8 solution were mixed, and the mixture was
then added into each plate and continued culture for another 2 h.
Next, the OD value at 450 nm was read using a microplate reader (Synergy
TM2, BIO-TEK Instruments Inc.). For in vitro PTT, HCT-116 cells were
incubated in 96-well plates and cultured for 12 h. Then, the dispersion
containing Hu-ink (25 or 50 μg/mL) was added into each plate
and incubated for another 2 h, followed by an 808 nm laser irradiation
(2 W/cm2, 5 min). Next, the CCK-8 assay was used to test
relative cell viabilities. For further qualitative assessment of PTT,
HCT-116 cells (2.0 × 105 cells per well) were seeded
in a 20 mm glass-bottom culture dish (NEST, China), cultured for 2
h with 25 or 50 μg/mL Hu-ink at 37 °C, and subsequently
followed by an 808 nm laser irradiation (2 W/cm2, 5 min).
Finally, the cells were stained for 20 min with PI and calcein AM
and observed by a ZEISS LSM710 live cell confocal laser imaging system
(Carl Zeiss, Germany).
of Hu-ink was assessed by CCK-8 assay. Approximately 1 × 104 HCT-116 and SW-620 cells were plated in 96-well plates and
cultured for 24 h at the standard cell culture environment. The cells
were cultured with samples (0–200 μg/mL) for 24 h and
then washed twice with PBS; 100 μL of RPMI 1640 cell medium
and 10 μL of CCK-8 solution were mixed, and the mixture was
then added into each plate and continued culture for another 2 h.
Next, the OD value at 450 nm was read using a microplate reader (Synergy
TM2, BIO-TEK Instruments Inc.). For in vitro PTT, HCT-116 cells were
incubated in 96-well plates and cultured for 12 h. Then, the dispersion
containing Hu-ink (25 or 50 μg/mL) was added into each plate
and incubated for another 2 h, followed by an 808 nm laser irradiation
(2 W/cm2, 5 min). Next, the CCK-8 assay was used to test
relative cell viabilities. For further qualitative assessment of PTT,
HCT-116 cells (2.0 × 105 cells per well) were seeded
in a 20 mm glass-bottom culture dish (NEST, China), cultured for 2
h with 25 or 50 μg/mL Hu-ink at 37 °C, and subsequently
followed by an 808 nm laser irradiation (2 W/cm2, 5 min).
Finally, the cells were stained for 20 min with PI and calcein AM
and observed by a ZEISS LSM710 live cell confocal laser imaging system
(Carl Zeiss, Germany).