Figure 2
(A) Alizarin red stain was added into cell cultures in osteogenic medium after photobiomodulation at a dose of 3 J/cm2 five times (every two days) with or without CPZ or SKF pretreatment. The alizarin red staining was measured after 21 days to determine the level of mineralization. Pre-incubation with CPZ (5 μM) and SKF (5 μM) for 10 minutes before photobiomodulation reduced the effect of photobiomodulation in 420 nm and 540 nm groups, to a lesser extent in the 660 nm group, but not in the 810 nm group. (B) Images of alizarin red staining taken by microscope. A higher intensity of alizarin red after 420 nm and 540 nm groups, while the intensity of 420 nm and 540 nm +CPZ/SKF groups was similar to the control group. (C,D) Quantitative evaluation of calcium deposits using Alizarin red staining. hASCs were treated or not with the TRP channel inhibitors CPZ (5 μM) and SKF (5 μM) for 10 minutes before each application of photobiomodulation. Data are expressed as mean ± SD. Experiments have been carried out for 3 times (n = 8). *,#P < 0.05, ***,###P < 0.001.
