Photodynamic Therapy is based on the activation of a photosensitizer by light in the presence of oxygen, generating reactive oxygen species that cause cellular destruction. One of the limitations of Photodynamic Therapy is the loss of capacity to absorb light (photobleaching), to generate reactive species, and thus, reducing the efficiency of the treatment.
In this work, the kinetics of photobleaching of methylene blue was studied, in a medium that simulates biological tissues (0.1mg/ml of collagen hydrolyzed in milli-Q water). The samples were irradiated in the presence of methylene blue (50 μM, 62.5 μM, 75 μM, and 100 μM); the transmission of a laser beam (Initial power (P0) = 7mW and wavelength (λ) = 635nm) through the samples was measured and the photobleaching kinetic rates (K) were calculated. In addition, videos of each sample were recorded simultaneously during the photobleaching process and the optical flow was demonstrated using the Python programming language.
It was observed that the kinetic rates of methylene blue photobleaching were decreasing as the methylene blue concentration increases. Thus, it was found that the microenvironment in which methylene blue is located influences its photobleaching.