## 4 Discussion
Successful odontogenic differentiation also depends on the choice of the stem cell source, culture conditions, and the presence of the right growth factors or signaling molecules [12; 62]. LLLT has been investigated for its potential to promote the osteeo/odorogenic differentiation of stem cells, such as DPSCs [63], SCAP [57], and PDLSCs [64]. GMSCs were found to form calcified nodules with positive Alizarin Red S staining in both experimental groups. The quantitative absorption values of the Alizarin Red S staining revealed a significantly higher value in group I than in group II. Once again, the lower energy dose caused a better biological response by GMSCs for odontogenic differentiation. Different energy doses of laser irradiation can activate other signaling pathways within cells. Lower energy doses may selectively activate specific pathways involved in odontogenic differentiation, leading to a more focused and efficient cellular response. Higher energy doses, on the other hand, may activate additional or different pathways that could interfere with or dampen the desired odontogenic differentiation [65; 66]. During the osteogenic and odontogenic differentiation process, GMSCs experience changes in the patterns of gene expression that they exhibit, including the upregulation of certain markers. The expression of osteogenic and odontogenic genes was analyzed using quantitative real-time PCR for a deeper evaluation of the osteeo/odontogenic differentiation. Both experimental groups showed significant levels of gene expression, which was indicative of the commitment and maturation of GMSCs in the direction of the osteoblast and odontoblast lineages. When compared to the control and osteogenic groups, the expression of Runt-related transcription factor 2 (Runx2), which is one of the earliest markers of osteogenic differentiation and plays a crucial role in the commitment of mesenchymal stem cells to the osteoblast lineage [67], was significantly increased in both laser groups. This contrasted with the groups that received osteogenic treatment. However, a decrease in Runx2 expression was observed in the high-energy group (3 J/cm\({}^{2}\)), which aligns with previous findings. The expression of Runx2 was significantly increased following laser irradiation at 0.4 and 1.9 J/cm\({}^{2}\), as demonstrated in previous research. However, a decrease in Runx2 expression in osteoblasts was found after using a laser at 3 J/cm\({}^{2}\), which was consistent with the current study in which group II had significantly lower expression of RUNX2 than group I. In this study, high upregulation of osteocalcin (OC) in groups I and II, compared to the control group and osteogenic group (positive control), indicated the maturation of osteoblasts. OC is a protein that mature osteoblasts produce and is associated with their terminal differentiation [68]. Previous studies have shown an upregulation of OC following LLLT at different energy densities [69; 70]. [PERSON] et al. [71] applied LLLT with different wavelengths and energy densities to BMMSCs and found that LLLT promoted osteogenesis. [PERSON] et al. [72] demonstrated that LLLT significantly increased the expression of the osteogenesis-related genes RUNX2, ALP, and OC at low energy levels of LLLT. The authors also revealed that the inhibition of the osteogenic markers was evident after using high-power energy densities, which agrees with the present results. Dentin sialophosphoprotein (DSPP) is thought to be a unique marker for odontogenic differentiation [73], and it plays a vital role in the regeneration of dentin. Both bone and dentin contain DSPP in varying amounts. However, dentin contains a substantially larger concentration of it than bone. Dentin matrix protein-1 (DMP1) is yet another protein that is essential to the process of odontogenesis and has a significant impact on the mineralization of dentin [74].