Stomatological Hospital of Chongqing Medical University, Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing 404100, China; Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing 404100, China.
Stomatological Hospital of Chongqing Medical University, Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing 404100, China; Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing 404100, China.
Int Immunopharmacol. 2022 Jul;108:108757. doi: 10.1016/j.intimp.2022.108757. Epub 2022 Apr 15.
All-trans retinoic acid (ATRA), a main derivative of vitamin A, has been shown to affect the osteogenic differentiation of mesenchymal stem cells (MSCs). Periodontal ligament stem cells (PDLSCs) possess characteristics of MSC and show strong potential for use in periodontal tissue restoration. However, the effect of ATRA on the osteogenic differentiation of PDLSCs remains unclear. In this study, we explored the effect of ATRA on the PDLSCs osteogenic differentiation. PDLSCs were harvested from the periodontalmembrane and treated with or without ATRA. CCK-8 and cell cycle analysis were used to evaluate PDLSC proliferation. PDLSC migration was assessed by scratch tests. qRT-PCR, western blotting, alkaline phosphatase staining, alizarin red staining and calcium quantification were performed to estimate the PDLSCs osteogenic differentiation capability and RNA sequencing to select differentially expressed genes (DEGs). Expression and activation of signaling elements were assessed by qRT-PCR, western blotting and immunofluorescence. Finally, we discovered that ATRA repressed the migration, proliferation, and osteogenesis ability of PDLSCs. RNA sequencing revealed 493 DEGs. Levels of interleukin-1β (IL-1β) were increased at varied time points after ATRA treatment. The inhibitive influence of ATRA on the osteogenesis of PDLSCs was partially reversed after neutralizing IL-1β. In addition, IL-1β levels were significantly attenuated by nuclear factor-κB (NF-κB) inhibitor BAY11-7082 and NLRP3 inhibitor MCC950. Taken together, our results demonstrate that ATRA disrupts the osteogenesis and mineralizationof PDLSCs by promoting IL-1β expression via activating NF-κB signaling and NLRP3 inflammasome, which may offer a new method for improving the ATRA-induced disruption of osteoblast differentiation.
全反式视黄酸（ATRA）是维生素 A 的主要衍生物之一，已被证明可影响间充质干细胞（MSCs）的成骨分化。牙周膜干细胞（PDLSCs）具有 MSC 的特征，并显示出在牙周组织修复中具有很强的应用潜力。然而，ATRA 对 PDLSCs 成骨分化的影响尚不清楚。在这项研究中，我们探讨了 ATRA 对 PDLSCs 成骨分化的影响。从牙周膜中分离出 PDLSCs，并在有无 ATRA 的情况下进行处理。使用 CCK-8 和细胞周期分析来评估 PDLSC 的增殖。通过划痕实验评估 PDLSC 的迁移。通过 qRT-PCR、western blot、碱性磷酸酶染色、茜素红染色和钙定量来评估 PDLSCs 的成骨分化能力，并通过 RNA 测序来选择差异表达基因（DEGs）。通过 qRT-PCR、western blot 和免疫荧光评估信号元素的表达和激活。最后，我们发现 ATRA 抑制了 PDLSCs 的迁移、增殖和成骨能力。RNA 测序显示有 493 个 DEGs。在 ATRA 处理后的不同时间点，白细胞介素-1β（IL-1β）的水平增加。中和 IL-1β 后，ATRA 对 PDLSCs 成骨的抑制作用部分逆转。此外，NF-κB 抑制剂 BAY11-7082 和 NLRP3 抑制剂 MCC950 显著降低了 IL-1β 的水平。综上所述，我们的研究结果表明，ATRA 通过激活 NF-κB 信号和 NLRP3 炎性小体促进 IL-1β 的表达，从而破坏 PDLSCs 的成骨和矿化，这可能为改善 ATRA 诱导的成骨细胞分化破坏提供新方法。
