Persistent Exposure to Fusobacterium nucleatum Triggers Chemokine/Cytokine Release and Inhibits the Proliferation and Osteogenic Differentiation Capabilities of Human Gingiva-Derived Mesenchymal Stem Cells
Persistent Exposure to Fusobacterium nucleatum Triggers Chemokine/Cytokine Release and Inhibits the Proliferation and Osteogenic Differentiation Capabilities of Human Gingiva-Derived Mesenchymal Stem Cells
复制标题
持续暴露于具核梭杆菌会触发趋化因子/细胞因子释放并抑制人牙龈源性间充质干细胞的增殖和成骨分化能力
DOI：
10.3389/fcimb.2019.00429
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发表时间：
2019-12-17
影响因子：
5.7
通讯作者：
Feng, Qiang
中科院分区：
文献类型：
--
作者：
Kang, Wenyan;Ji, Xiaoli;Feng, Qiang
具核梭杆菌是引起牙周炎最常见的致病菌之一。具核梭杆菌（Fusobacterium nucleatum，F. nucleatum）对口腔干细胞的影响很少报道。在这项研究中，我们的目的是评估牙龈来源的间充质干细胞（GMSC）如何应对直接挑战与F。核质采用有限稀释法分离骨髓间充质干细胞，并将其暴露于F.在不同的感染复数（MOI; F.核：细胞比率为10：1、50：1和100：1）培养24小时至4周。结果表明，F.具核质以剂量依赖性方式显著抑制细胞增殖，并促进细胞迁移和趋化因子/细胞因子如CCL 2、CXCL 1和IL-6的释放。此外，F.核质抑制GMSC成骨分化部分是通过降低碱性磷酸酶（ALP）活性、矿化结节形成以及成骨相关基因和蛋白表达。RNA测序分析表明，F.成骨细胞分化过程中，核仁时间依赖性地激活细胞信号通路。共发现64个细胞分化相关基因在未感染和F.在第3、7、14和21天，有趣的是，我们发现64个细胞分化相关的差异表达基因（DEG）在癌症相关通路中显著富集，如骨癌、骨肉瘤和骨髓癌，这为GMSC成骨分化过程中的肿瘤发生提供了新的见解。总之，本研究表明，持续暴露于F。核质促进细胞迁移和趋化因子/细胞因子释放，并抑制GMSC的增殖和成骨分化。本研究提供了一种新的、长时间的细菌-细胞体外共培养模型，为进一步研究F.核质感染
Fusobacterium nucleatum is one of the most frequent pathogenic bacteria causing periodontitis. The direct effect of Fusobacterium nucleatum (F. nucleatum) on oral stem cells has rarely been reported. In this study, we aimed to evaluate how gingiva-derived mesenchymal stem cells (GMSCs) respond to a direct challenge with F. nucleatum. GMSCs were isolated by the limiting dilution method and exposed to F. nucleatum at various multiplicities of infection (MOIs; F. nucleatum:cell ratios of 10:1, 50:1, and 100:1) for 24 h to 4 weeks. Our results indicated that F. nucleatum significantly inhibited cell proliferation in a dose-dependent manner and promoted cell migration and the release of chemokines/cytokines, such as CCL2, CXCL1, and IL-6. Additionally, F. nucleatum inhibited GMSC osteogenic differentiation partly by decreasing alkaline phosphatase (ALP) activity, mineralized nodule formation, and osteogenesis-related gene and protein expression. RNA-sequencing analyses indicated that F. nucleatum time-dependently activated cellular signaling pathways during the process of osteogenic differentiation. A total of 64 cell differentiation-related genes were found to be differentially expressed between non-infected and F. nucleatum-infected GMSCs at 3, 7, 14, and 21 d. Intriguingly, we discovered that the 64 cell differentiation-related differentially expressed genes (DEGs) were significantly enriched in cancer-related pathways, such as bone cancer, osteosarcoma and bone marrow cancer, which provides new insight into tumorigenesis during the process of GMSC osteogenic differentiation. In conclusion, this study demonstrates that persistent exposure to F. nucleatum promotes cell migration and chemokine/cytokine release and inhibits the proliferation and osteogenic differentiation of GMSCs. Our study provides a novel and long-time bacteria-cell co-culture in vitro model and makes a foundation for the future mechanistic studies of GMSCs under F. nucleatum infection.