Tensile stress induced osteogenesis of periodontal ligament cells via Piezo1 mediated TAZ-Cbfα1 signaling.

Sisi Li, Ting Jin, Yi Wang, Hui Deng, Rongdang Hu
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Abstract

Objective: 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.

Design: 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.

Results: 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.

Conclusions: 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.

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目的:已知循环拉伸应力(CTS)可诱导牙周韧带细胞(PDLCs)成骨,其分子机制尚待阐明。本研究旨在探讨机械敏感性钙通道Piezo1在拉伸应变下PDLCs成骨过程中的作用,以及TAZ-Cbfα1通路在此过程中的信号调控:设计:从牙周韧带组织中分离出PDLCs,并对其进行CTS处理。采用茜素红染色法(ARS)和碱性磷酸酶(ALP)检测PDLCs的成骨过程。RT-qPCR和Western印迹法分别检测Piezo1、具有PDZ结合基调的转录共激活因子(TAZ)和核心结合因子α1(Cbfα1)的转录本和蛋白表达水平。免疫荧光染色用于检测 TAZ 的核聚集。采用靶向Piezo1的小干扰RNA(siRNA)抑制Piezo1 mRNA的表达:结果表明:在CTS作用下,PDLCs的成骨分化能力显著增强,Piezo1、TAZ和Cbfα1的mRNA和蛋白表达水平升高。此外,ALP活性和ARS染色钙结节的形成也明显增加。此外,Piezo1 siRNA感染可明显抑制CTS诱导的TAZ-Cbfα1通路和PDLCs的成骨过程,表明Piezo1具有调控作用:我们提供的证据表明,应用CTS可促进PDLCs的成骨分化,而这可能是由Piezo1靶向TAZ-Cbfα1信号传导介导的。
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