Effect of photobiomodulation on the behaviour of mesenchymal stem cells in three-dimensional cultures.

Photobiomodulation (PBM) has been proposed as a strategy to improve the regenerative capacity of human adipose-derived stem cells (hASCs). Yet, this effect has been proved in 2D culture conditions. To analyze the effect of different doses of laser irradiation (660 nm) with different levels of energy (1 J, 2 J and 6 J) on hASCs cultured at 2D and 3D conditions. We used gellan gum spongy-like hydrogels as a biomaterial to 3D culture hASCs. Different doses (1-7 daily irradiations) and energy levels (1-6 J) of PBM were applied, and the metabolic activity, viability, proliferation, and release of ROS and IL-8 was evaluated up to 7 days. In 3D, cell proliferation increased at high energy (6 J) and after a single dose of irradiation, while in 2D, metabolic activity and proliferation was enhanced only after 3 doses and independently of the energy. More than 1 dose was needed to promote ROS secretion both in 2D and 3D culture conditions. Interestingly, a decrease of IL-8 secretion was detected only in 3D after 3-7 daily irradiations. Overall, hASCs response to PBM was not only dependent on the energy level and the number of applied stimuli, but also on the in vitro culture conditions.