Home > Browse Issues > Vol.40 No.9
Mechanically Sensitive Ion Channels Trpm7 in LIPUS Promotes Interdental Dentinal Mesenchyme Study on the Mechanism of Action in Osteogenic Differentiation
Yao Huan, Pan Huafeng, Zheng Yan, Zhang Jie, Li Yasha, Wu Mengyun, Yang Ke*
Key Laboratory of Children’s Developmental Diseases Research, Ministry of Education, Children’s Hospital of Chongqing Medical University; National International Science and Technology Cooperation Base for Major Children’s Development; Chongqing Key Laboratory of Pediatrics; Chongqing Stem Cell Therapy Engineering Research Center, Chongqing Cell Bioengineering Technology Co., Ltd., Chongqing 400014, China
Abstract: The purpose of this study was to investigate the role of low-intensity pulsed ultrasound (LIPUS) in promoting osteogenic differentiation of dental pulp mesenchymal stem cells (DPSCs) and the role of transient receptor potential M7 (Transient receptor potential melastatin 7， Trpm7) involved. The human DPSCs were cultured. The surface molecular markers were detected by flow cytometry. The alcian blue， alizarin red and oil red O staining were used to detect the chondrogenic， osteogenic and adipogenic differentiation ability. ALP activity， ALP staining and alizarin red staining were observed the ability of LIPUS to promote osteogenic differentiation. The Real-time quantitative PCR was used to detect the difference of OPN， OCN and RUNX2 expression in the LIPUS-treated group and the control group， and the mRNA expression levels of Trpm7 in the two groups were detected at different time points. ALP activity was used to examine the effect of LIPUS and different concentrations of Trpm7 inhibitor 2-Aminoethoxydiphenyl borate (2-APB) on osteogenic differentiation. The osteogenic differentiation ability was observed by ALP staining and alizarin red staining in control group， LIPUS group， LIPUS+DMSO group and LIPUS+2-APB group. The expressions of OPN， OCN and RUNX2 were detected by Western blot. The results showed the cultured cells were identified as DPSCs. In the LIPUS treatment group， ALP staining and alizarin red staining increased significantly， and the ALP activity enhanced (P<0.01). The mRNA expression of OPN， OCN and RUNX2 were significantly increased (P<0.05). The mRNA expression of Trpm7 was significantly increased on the 5th day (P<0.05). 2-APB significantly inhibited ALP activity (P<0.01). Compared with the control group， ALP staining， alizarin red staining and the protein expression of OPN， OCN， RUNX2 were significantly increased in LIPUS group and LIPUS+DMSO group. Compared to LIPUS+DMSO group， ALP and alizarin red staining， the protein expression of OPN， OCN， RUNX2 were significantly reduced in LIPUS+2-APB group. We concluded that LIPUS can promote osteogenic differentiation of DPSCs， and the mechanically sensitive ion channel Trpm7 plays an important role in this process.
CN
EN