Investigating the Osteoinductive Effect of Shilajit on the Differentiation of Rat Bone Marrow Mesenchymal Stem Cells into Osteoblasts

Introduction: Bone marrow–derived mesenchymal stem cells (BMSCs) are considered a promising cell source for tissue engineering and bone regeneration. Previous studies have indicated that large bone defects can be effectively repaired using autologous mesenchymal stem cells. This study aimed to evaluate the effect of the natural bioactive compound Shilajit on the osteogenic differentiation of BMSCs. Methods: BMSCs were isolated from rat bone marrow using the flushing-out technique. At the fourth passage, cells were allocated into seven groups, including a control group and six experimental groups. Experimental groups were treated with Shilajit at the concentrations of 150, 300, and 600 mg/mL, with or without osteogenic differentiation medium, for 21 days. Cell viability, alkaline phosphatase (ALP) activity, and Alizarin Red staining were assessed to evaluate proliferation and osteogenic differentiation. Data were analyzed using one-way ANOVA, and differences were considered statistically significant at P<0.05. Results: All Shilajit-treated groups exhibited a significant increase in ALP activity and calcium deposition, as evidenced by Alizarin Red staining, compared to the control group. The highest ALP activity was detected in cells treated with 300 mg/mL Shilajit in combination with osteogenic differentiation medium. Conclusion: The findings suggest that Shilajit enhances the osteogenic differentiation of BMSCs, particularly at a concentration of 300 mg/mL. These results highlight the potential of Shilajit as a bioactive compound for promoting bone tissue regeneration; however, further in vivo and clinical studies are warranted to confirm its efficacy and safety in bone repair applications.