Objectives: Many studies have demonstrated that the clock gene, brain and muscle ARNT-like 1 (Bmal1), is directly related to bone ageing by affecting age-related changes to mesenchymal stem cells (MSCs). As a main developmental signal, Wnt may play an important role in this process. Here, we have aimed to elucidate whether Bmal1 positively regulates osteogenesi via Wnt pathways.
Materials and methods: Bone marrow stromal cells were cultured in basic and in osteo-induction medium with Wnt signalling inhibitor Dkk1 and Bmal1 transfection. Proliferation and osteogenesis of MSCs, expression of Bmal1 and activation of Wnt signalling were investigated by flow cytometry, senescence-associated β-galactosidase (SA-β-gal) staining, real-time quantitative PCR and western blot analysis.
Results: Expression of Bmal1 (specially after 7 days osteo-induction), activation of Wnt signalling and osteo-related factors fell significantly during osteo-induction after Dkk1 addition. When cellular Bmal1 was increased by transfection, osteogenesis inhibition by Dkk1 was rescued to a certain extent with activation of Wnt signalling. However, Dkk1 did not significantly affect proliferation or senescence of MSCs during early periods of culture.
Conclusion: These findings demonstrated that Bmal1 and Wnt signalling may have a synergistic effect at a particular stage of osteogenesis. Inhibition of Wnt signalling did not greatly affect ageing of MSCs through early passages.
© 2013 John Wiley & Sons Ltd.