Li Sisi, Jin Ting, Wang Yi, Deng Hui, Hu Rongdang
Department of Orthodontics, School and Hospital of Stomatology, Wenzhou Medical University, Wenzhou, Zhejiang, China; Department of Stomatology, The Third Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China.
Department of Orthodontics, School and Hospital of Stomatology, Wenzhou Medical University, Wenzhou, Zhejiang, China; Department of Stomatology, Taizhou Hospital of Zhejiang Province affiliated to Wenzhou Medical University, Enze Hospital, Taizhou, Zhejiang, China.
Arch Oral Biol. 2025 Mar;171:106152. doi: 10.1016/j.archoralbio.2024.106152. Epub 2024 Nov 29.
Cyclic tensile stress (CTS) is known to induce osteogenesis of periodontal ligament cells (PDLCs), in which the molecular mechanism remains to be elucidated. This study aimed to investigate the role of the mechanosensitive calcium channel Piezo1 in the osteogenesis of PDLCs under tensile strain, as well as the signal regulation of the TAZ-Cbfα1 pathway in this process.
PDLCs were isolated from periodontal ligament tissues and subjected to CTS. Alizarin red staining (ARS) and alkaline phosphatase (ALP) assay were used to detect the osteogenesis of PDLCs. RT-qPCR and Western blot were used to detect the transcripts and protein expression levels of Piezo1, Transcriptional co-activator with PDZ binding motif (TAZ), and Core-binding factor α1 (Cbfα1) respectively. Immunofluorescence staining was used to detect the nuclear aggregation of TAZ. Small interfering RNA (siRNA) targeting Piezo1 (Piezo1-siRNA) was adopted to inhibit Piezo1 mRNA expression.
The results showed that the osteogenic differentiation capacity of PDLCs was significantly enhanced under CTS, along with elevated mRNA and protein expression levels of Piezo1, TAZ, and Cbfα1. Moreover, the ALP activity and the formation of calcium nodules by ARS staining were significantly increased. In addition, Piezo1 siRNA infection significantly inhibited the CTS-induced TAZ-Cbfα1 pathway and the osteogenesis of PDLCs, suggesting the regulatory role of Piezo1.
We provided evidence that the application of CTS encourages the osteogenic differentiation of PDLCs, which could be mediated by the Piezo1 targeted TAZ-Cbfα1 signaling.
已知循环拉伸应力(CTS)可诱导牙周膜细胞(PDLCs)成骨,但其分子机制仍有待阐明。本研究旨在探讨机械敏感钙通道Piezo1在拉伸应变下对PDLCs成骨的作用,以及该过程中TAZ-Cbfα1信号通路的调控机制。
从牙周膜组织中分离出PDLCs,并对其施加CTS。采用茜素红染色(ARS)和碱性磷酸酶(ALP)检测法检测PDLCs的成骨情况。分别运用RT-qPCR和蛋白质免疫印迹法检测Piezo1、具有PDZ结合基序的转录共激活因子(TAZ)和核心结合因子α1(Cbfα1)的转录本和蛋白表达水平。采用免疫荧光染色法检测TAZ的核聚集情况。采用靶向Piezo1的小干扰RNA(siRNA)(Piezo1-siRNA)抑制Piezo1 mRNA表达。
结果显示,在CTS作用下,PDLCs的成骨分化能力显著增强,同时Piezo1、TAZ和Cbfα1的mRNA和蛋白表达水平升高。此外,ALP活性及ARS染色显示的钙结节形成显著增加。此外,Piezo1 siRNA感染显著抑制了CTS诱导的TAZ-Cbfα1信号通路及PDLCs的成骨作用,提示Piezo1具有调控作用。
我们提供的证据表明,CTS的应用促进了PDLCs的成骨分化,这可能由Piezo1靶向的TAZ-Cbfα1信号介导。