Midazolam enhances osteogenic differentiation of human bone marrow mesenchymal stem cells

Background: Bone remodeling involves a balance between osteoblast-driven formation and osteoclast-mediated resorption, with disruptions leading to diseases like osteoporosis. Midazolam (MDZ), known for its sedative properties, has shown effects on cellular differentiation and hydroxyapatite formation in dental cells. However, its role in promoting osteogenic differentiation in human bone marrow mesenchymal stem cells (hBMSCs) remains unexplored, motivating this study to investigate its potential in bone regenerative therapies. Methods: Human bone marrow stem cells were cultured in the complete media with α-MEM, 10% FBS, and 1% pen/strep. Cell viability was determined with MTT assay in different concentrations of MDZ (0.125 to 1 µM) for 72 hours. Osteogenic differentiation was induced over 21 days using the selected doses of MDZ with osteogenic medium. The Alizarin Red S (ARS) staining was performed to determine the calcium deposit for osteoblast cells. Data were analyzed using repeated measure ANOVA, and a p-value <0.05 was considered statistically significant. Results: The MTT results for different concentrations of MDZ (0.125 to 1 µM) showed no significant cytotoxic effects on hBMSCs after 72 hours. Furthermore, ARS staining revealed increased calcium deposits in 0.5 µM MDZ compared to untreated and DMSO groups (P-Value =0.0001). These findings suggest that MDZ promotes mineralization at lower concentrations, highlighting its potential in osteogenic applications. Conclusion: Midazolam promotes osteogenic differentiation of hBMSCs, particularly at 0.5 µM concentration, without cytotoxic effects. These findings demonstrate that MDZ may be a potential compound for osteoblastogenesis; however, these findings require further in vivo studies to confirm the idea.