College of Stomatology, Chongqing Medical University; Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences; Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing 401147, P.R. China.
Int J Mol Med. 2018 Jul;42(1):322-330. doi: 10.3892/ijmm.2018.3592. Epub 2018 Mar 27.
Low‑intensity pulsed ultrasound (LIPUS) is a non‑invasive therapeutic treatment for accelerating fracture healing. A previous study from our group demonstrated that LIPUS has the potential to promote periodontal tissue regeneration. However, the underlying molecular mechanism by which LIPUS promotes periodontal tissue regeneration remains unknown. In the present study, periodontal ligament stem cells (PDLSCs) were isolated from premolars. Flow cytometry and differentiation assays were used to characterize the isolated PDLSCs. LIPUS treatment was administered to PDLSCs, and stromal cell‑derived factor‑1 (SDF‑1) expression levels were examined by reverse transcription‑quantitative polymerase chain reaction with or without blocking the SDF‑1/C‑X‑C motif chemokine receptor 4 (CXCR4) pathway with AMD3100. ELISA was used to evaluate SDF‑1 secretion in PDLSCs. Wound healing and transwell assays were conducted to assess the migration‑promoting effect of LIPUS. A potential upstream gene of SDF‑1, twist family bHLH transcription factor 1 (TWIST1), was silenced by small interfering (si) RNA transfection. The results demonstrated that LIPUS treatment promoted the expression of TWIST1 and SDF‑1 at both the mRNA and protein levels. In addition, LIPUS treatment enhanced the cell migration of PDLSCs. Knockdown of TWIST1 impaired the expression of SDF‑1 and the cell migration ability of PDLSCs. TWIST1 may be an upstream regulator of SDF‑1 in PDLSCs. Taken together, these findings indicate that the SDF1/CXCR4 signaling pathway is involved in LIPUS‑promoted PDLSC migration, which might be one of the mechanisms for LIPUS‑mediated periodontal regeneration. TWIST1 might be a mechanical stress sensor during mechanotransduction.
低强度脉冲超声(LIPUS)是一种促进骨折愈合的非侵入性治疗方法。我们小组的先前研究表明,LIPUS 具有促进牙周组织再生的潜力。然而,LIPUS 促进牙周组织再生的潜在分子机制尚不清楚。在本研究中,从前磨牙中分离出牙周韧带干细胞(PDLSCs)。通过流式细胞术和分化实验对分离的 PDLSCs 进行了表征。对 PDLSCs 进行 LIPUS 处理,并通过逆转录定量聚合酶链反应检查基质细胞衍生因子-1(SDF-1)的表达水平,同时用 AMD3100 阻断 SDF-1/C-X-C 基序趋化因子受体 4(CXCR4)途径。ELISA 用于评估 PDLSCs 中 SDF-1 的分泌。进行伤口愈合和 Transwell 测定以评估 LIPUS 的促迁移作用。通过小干扰(si)RNA 转染沉默 SDF-1 的潜在上游基因,卷曲螺旋-碱性螺旋-环-螺旋转录因子 1(TWIST1)。结果表明,LIPUS 处理促进了 TWIST1 和 SDF-1 在 mRNA 和蛋白质水平上的表达。此外,LIPUS 处理增强了 PDLSCs 的细胞迁移。TWIST1 的敲低削弱了 SDF-1 的表达和 PDLSCs 的细胞迁移能力。TWIST1 可能是 PDLSCs 中 SDF-1 的上游调节剂。总之,这些发现表明 SDF1/CXCR4 信号通路参与了 LIPUS 促进 PDLSC 迁移,这可能是 LIPUS 介导牙周再生的机制之一。TWIST1 可能是机械转导过程中的机械应力传感器。