Abstract
Biodentine is widely used as a pulp capping agent due to its biocompatibility and ability to stimulate reparative dentin formation. Considering the growing use of lasers in dental clinics, this study aimed to assess the effects of 450 and 980 nm diode laser irradiations combined with Biodentine on proliferation and the expression of pluripotent genes of dental pulp stem cells (DPSCs). DPSCs were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FBS. The experiment consisted of ten groups, each subjected to different conditions of diode laser irradiation at two wavelengths (450 nm and 980 nm) and energy densities (2 J/cm² and 4 J/cm²), with or without the addition of Biodentine extract. The groups included controls such as Biodentine extract alone and untreated DPSCs. Cell proliferation was assessed using MTT assays. The expression of pluripotent genes (OCT4, SOX2, NESTIN, and NANOG) was evaluated through real-time PCR. Data were analyzed by two-way ANOVA (P < 0.05). DPSCs exposed to 450 nm /4 J/cm² in combination of Biodentine showed significantly higher proliferation when compared to the Biodentine group at 24, 48, and 72 h (P < 0.005). Moreover, the 450 nm/4 J/cm² laser induced the highest expression of pluripotent genes in cells exposed to Biodentine, whereas the 980 nm/4 J/cm² laser group demonstrated the lowest gene expression. Application of Biodentine in combination with photobiomodulation (PBM) therapy can enhance DPSC proliferation and increase the expression of pluripotent genes. PBM may be optimized as an adjunctive treatment to Biodentine in regenerative endodontics, promoting more effective tissue regeneration and healing. However, this study was limited to in vitro conditions, without in vivo validation or assessment of mineralization potential, highlighting the need for further studies to confirm these findings in clinical settings.