Oral and Maxillofacial Reconstruction and Regeneration Laboratory, Southwest Medical University, Luzhou 646000, China.
Oral and Maxillofacial Reconstruction and Regeneration Laboratory, Southwest Medical University, Luzhou 646000, China; Department of Oral and Maxillofacial Surgery, The Affiliated Stomatology Hospital of Southwest Medical University, Luzhou 646000, China; Department of Oral and Maxillofacial Surgery, The Affiliated Hospital of Southwest Medical University, Luzhou 646000, China.
Ann Anat. 2022 Jan;239:151847. doi: 10.1016/j.aanat.2021.151847. Epub 2021 Oct 20.
To explore the role of the mechanosensitive ion channel Piezo1 in the proliferation and osteogenic differentiation of human dental follicle cells (hDFCs), and its mechanism, so as to provide the basis for the use of hDFCs to achieve bone regeneration.
hDFCs were obtained from fresh dental follicle tissues by enzymatic digestion, and cell phenotype and multipotential differentiation were identified. Identification of the expression of mechanosensitive ion channel Piezo1 was performed by immunofluorescence and immunohistochemistry. CCK-8 was used to determine the optimal concentration of the Piezo1 agonist, Yoda1. Then, according to the obtained results, Alizarin red staining, RT-PCR quantitative analysis and Western blot were used to further observe the osteogenic differentiation of hDFCs and its probable mechanism via Wnt/β-catenin signalling. The data were analysed by SPSS 22.0 software.
The results of the concentration gradient experiments indicated that 0.5 µM Piezo1 agonist (Yoda1) enhanced the proliferation of hDFCs. Compared with the control group, a considerable number of calcium nodules showed that activating Piezo1 could promote the osteogenic differentiation of hDFCs. The relative mRNA and protein expression of Piezo1, ALP, RUNX2, OCN and BMP2 in the Piezo1 agonist group were higher than that of the control group. Furthermore, the expression of Wnt3a and β-catenin related to the classical osteogenic pathway were significantly up-regulated in the Piezo1 agonist group.
Activating mechanosensitive ion channel Piezo1 with an appropriate concentration of Yoda1 has a positive effect on the proliferation and osteogenic differentiation of hDFCs. This mechanism of promoting osteogenic differentiation may be mediated by the Wnt/β-catenin pathway.
为了探讨机械敏感离子通道 Piezo1 在人牙周膜细胞（hDFCs）增殖和成骨分化中的作用及其机制，为利用 hDFCs 实现骨再生提供依据。
酶消化法从新鲜牙周膜组织中获取 hDFCs，鉴定细胞表型和多能分化。免疫荧光和免疫组织化学鉴定机械敏感离子通道 Piezo1 的表达。CCK-8 法确定 Piezo1 激动剂 Yoda1 的最佳浓度。然后，根据所得结果，通过 Alizarin 红染色、RT-PCR 定量分析和 Western blot 进一步观察 hDFCs 的成骨分化及其通过 Wnt/β-catenin 信号通路的可能机制。数据采用 SPSS 22.0 软件进行分析。
浓度梯度实验结果表明，0.5µM Piezo1 激动剂（Yoda1）增强了 hDFCs 的增殖。与对照组相比，大量钙结节表明激活 Piezo1 可以促进 hDFCs 的成骨分化。Piezo1 激动剂组 hDFCs 的 Piezo1、ALP、RUNX2、OCN 和 BMP2 的相对 mRNA 和蛋白表达均高于对照组。此外，Piezo1 激动剂组经典成骨途径相关的 Wnt3a 和 β-catenin 的表达明显上调。
用适当浓度的 Yoda1 激活机械敏感离子通道 Piezo1 对 hDFCs 的增殖和成骨分化有积极作用。这种促进成骨分化的机制可能是通过 Wnt/β-catenin 通路介导的。
