Exploring the Role of Photobiomodulation in Cellular Responses: Implications for Cancer and Alzheimer’s Disease

This project was intended to conduct experimental studies on the effects of two types of electromagnetic (EM) waves on living states with potential applications as novel therapeutic modalities. The first part of this study, conducted in collaboration with the University of Alberta, involved cancer research and specifically explored potential therapeutic applications of photobiomodulation (PBM). PBM involves the use of non-ionizing EM radiation in the range between infrared and ultraviolet to induce biological effects on cells, tissues and organisms. The devices used for the experiments were the Bioptron device, which emits a hyperpolarized light beam (HPL) at 40 mW/cm², and the Vielight NeuroPro device, which utilizes an infrared light beam at 60 mW/cm² with a 10 Hz frequency sweep. Three different cancer cell lines were used in the experiments, namely: PC3, HeLa, and MCF7. The study focused on analyzing cell viability, morphological changes, ATP production, and metabolic shifts. The first step of the experiment involved culturing the aforementioned cells under standard conditions to promote proliferation and obtain statistically significant data for analysis. Subsequently, part of the samples was exposed to HPL (via Bioptron), while the remaining samples were exposed to infrared light (via Vielight). After irradiation, cell viability was first assessed using the Alamar Blue assay, followed by the analysis of key cytoplasmic proteins: actin, tubulin, and mitochondrial morphological changes through immunofluorescence staining. Finally, ATP production and metabolic shifts were quantified using the Glycolysis/OXPHOS Assay Kit. The study highlighted a biphasic cellular response to light irradiation: short exposure reduced cell viability, suggesting initial stress, whereas longer exposure produced variable effects, promoting proliferation in some cases and cell death in others. The Bioptron device induced more pronounced cellular changes than the Vielight NeuroPro device, with morphological alterations such as cytoplasmic shrinkage and potential mitochondrial damage. Metabolic assessments indicated a shift in energy production pathways following irradiation. Some experimental sets showed increased glycolytic activity with reduced mitochondrial ATP production, while others exhibited the opposite trend. This metabolic reprogramming appeared to be influenced by both the irradiation conditions and the specific cell line. An interesting finding was the role of the culture medium, which seemed to mediate the effects of electromagnetic waves, suggesting a complex interaction between light, the extracellular environment, and cellular response. The results suggest the therapeutic potential of PBM and HPL but emphasize the need for precise optimization of treatment protocols. In the second part of the thesis, conducted in collaboration with the University of Bologna, experiments were performed to study the effects of PBM on the Tau protein, which is involved in Alzheimer's disease (AD). The experiments were conducted on rat brain slices obtained using a Vibratome and preserved in artificial cerebrospinal fluid (ACSF) at 4°C to maintain slice viability throughout the process. Before applying the discussed therapy, a subset of samples underwent Tau protein hyperphosphorylation mimicking a key aspect of the AD through a gradual temperature reduction. The samples were analyzed using WesternBlot to assess the behavior of the Tau protein and its potential phosphorylation under the experimental conditions