Effect of red light and near infrared laser on the generation of reactive oxygen species in primary dermal fibroblasts

Irradiation with red light or near-infrared (NIR) lasers can bio-modulate cellular processes or revitalize injured tissues and therefore, widely been used for therapeutic interventions. Mechanistically, this cellular or biological process, referred as Photobiomodulation (PBM), is achieved by the generation of oxide free radicals in cells and tissues. This explorative study using red light (636 nm) and Near Infra-Red (NIR, 825 nm) laser at various irradiation exposures reckons the level of oxidative stress induced by these free radicals in human primary fibroblasts. Freshly isolated dermal fibroblasts were irradiated with red light and NIR at power densities of 74 and 104 mV/cm2, respectively and, at varying fluences ranging from 5 to 25 J/cm2. Cellular oxidative stress, measured by Reactive Oxygen Species (ROS) upon quantifying fluorescently labelled oxide free radicals in cells, detected considerable variations between the irradiation exposures of red light and NIR laser. The NIR laser demonstrated high levels of ROS at all fluences, except 10 J/cm2 indicating its ability in generating of two types of oxide radicals in dermal fibroblasts, often illustrated as biphasic response. Further, the responses of these cells to variable fluences of red light and NIR laser were measured to evaluate the immediate effect of PBM on cellular activity. The production of cellular energy coincides with the amount of oxidative stress, which was two-fold higher in cells irradiated with the NIR laser, as compared with the red light. This outcome indicates that the ROS production within biological systems are more dependent on the wavelength of the laser rather than its fluences. Further studies are required to avoid 'overdosing of PBM' and to analyse ROS qualitatively for making the best use of the red light and NIR laser in clinics.