Background: Bone mesenchymal stem cells (BMSCs) are recognized for their intrinsic capacity for self-renewal and differentiation into osteoblasts, adipocytes, and chondrocytes, making them pivotal entities within the field of bone research. Tescalcin (TESC) is known to play a role in specific cellular processes related to proliferation and differentiation. However, the precise involvement of TESC in the regulation of BMSCs remains unclear. The present study was designed to verify the functional implications of TESC in BMSCs.
Methods: An adenovirus vector was engineered to downregulate TESC expression, and the Wnt/β-catenin signaling pathway was activated using BML-284. The assessment of mRNA was conducted by quantitative real-time polymerase chain reaction (qRT-PCR). The assessment of protein expression was conducted by Western blotting and immunofluorescence techniques (IF), respectively. Alkaline phosphatase (ALP) staining and activity assays were performed to verify ALP changes, while Alizarin Red S (ARS) staining and quantitative analysis were employed to assess mineralization capacity.
Results: Initially, we observed an upregulation of TESC expression during osteogenic differentiation. Subsequently, TESC knockdown was demonstrated to decrease the osteogenic-related genes expression and diminish BMSCs mineralization. Concomitantly, we identified the inhibition of Wnt/β-catenin signaling following the TESC knockdown. Furthermore, the administration of BML-284 effectively activated the Wnt/β-catenin pathway, successfully rescuing the compromised TESC-mediated osteogenic differentiation.
Conclusion: Our findings indicate that TESC knockdown exerts an inhibitory effect on the osteogenic differentiation of BMSCs through the modulation of the Wnt/β-catenin signaling pathway. This study unveils a novel target with potential applications for enhancing the regenerative potential of BMSCs in the realm of regenerative medicine.
Keywords: BMSCs; TESC; Wnt/β-catenin signaling; marker; osteogenesis.
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