牙周膜细胞通过 miR-299-5p 调节间充质干细胞成骨。
Periodontal ligament cells regulate osteogenesis via miR-299-5p in mesenchymal stem cells.
机构信息
Department of Periodontal Medicine, Hiroshima University Graduate School of Biomedical and Health Sciences, Hiroshima, 734-8553, Japan.
Department of Periodontal Medicine, Hiroshima University Graduate School of Biomedical and Health Sciences, Hiroshima, 734-8553, Japan.
出版信息
Differentiation. 2020 Mar-Apr;112:47-57. doi: 10.1016/j.diff.2020.01.001. Epub 2020 Jan 9.
BACKGROUND
The periodontal ligament contains periodontal ligament cells, which is a heterogeneous cell population, and includes progenitor cells that can differentiate into osteoblasts/cementoblasts. Mesenchymal stem cells (MSCs) can differentiate into various cells and can be used for periodontal regenerative therapy. Therefore, transplanted MSCs can be affected by humoral factors from periodontal ligament cells via the transcription factors or microRNAs (miRNAs) of MSCs. In addition, periostin (POSTN) is secreted from HPL cells and can regulate periodontal regeneration and homeostasis. To clarify the regulatory mechanism of humoral factors from periodontal ligament cells, we attempted to identify key genes, specifically microRNAs, involved in this process.
METHODS
Human MSCs (hMSCs) were indirectly co-cultured with human periodontal ligament cells (HPL cells) and then evaluated for osteogenesis, undifferentiated MSCs markers, and miRNA profiles. Furthermore, hMSCs were indirectly co-cultured with HPL cells in the presence of anti-POSTN monoclonal antibody (anti-POSTN Ab) to block the effect of POSTN from HPL cells, and then evaluated for osteogenesis or undifferentiated MSC markers. Moreover, hMSCs showed alterations in miRNA expression or cultured with HPL were challenged with POSTN during osteogenesis, and cells were evaluated for osteogenesis or undifferentiated MSC markers.
RESULTS
hMSCs co-cultured with HPL cells showed suppressed osteogenesis and characteristic expression of SOX11, an undifferentiated MSC marker, as well as miR-299-5p. Overexpression of miR-299-5p regulated osteogenesis and SOX11 expression as observed with indirect co-culture with HPL cells. Furthermore, MSCs co-cultured with HPL cells were recovered from the suppression of osteogenesis and SOX11 mRNA expression by anti-POSTN Ab. However, POSTN induced miR-299-5p and SOX11 expression, and enhanced osteogenesis.
CONCLUSION
Humoral factors from HPL cells suppressed osteogenesis in hMSCs. The suppressive effect was mediated by miR-299-5p and SOX11 in hMSCs.
背景
牙周膜含有牙周膜细胞，这是一种异质细胞群体，包括可以分化为成骨细胞/成牙骨质细胞的祖细胞。间充质干细胞（MSCs）可以分化为各种细胞，并可用于牙周再生治疗。因此，移植的 MSCs 可能会受到牙周膜细胞的体液因子的影响，这些因子通过 MSCs 的转录因子或 microRNAs（miRNAs）起作用。此外，骨膜蛋白（POSTN）从 HPL 细胞分泌，可以调节牙周再生和稳态。为了阐明牙周膜细胞的体液因子的调节机制，我们试图鉴定参与该过程的关键基因，特别是 microRNAs。
方法
人 MSCs（hMSCs）与人类牙周膜细胞（HPL 细胞）间接共培养，然后评估成骨作用、未分化 MSC 标志物和 miRNA 谱。此外，hMSCs 在存在抗 POSTN 单克隆抗体（抗 POSTN Ab）的情况下与 HPL 细胞间接共培养，以阻断 HPL 细胞 POSTN 的作用，然后评估成骨作用或未分化 MSC 标志物。此外，hMSCs 的 miRNA 表达发生变化，或在成骨过程中与 HPL 共培养的细胞受到 POSTN 的刺激，然后评估其成骨作用或未分化 MSC 标志物。
结果
与 HPL 细胞共培养的 hMSCs 表现出成骨作用受抑制，以及未分化 MSC 标志物 SOX11 的特征表达，以及 miR-299-5p 的表达。miR-299-5p 的过表达调节成骨作用和 SOX11 的表达，这在与 HPL 细胞间接共培养时观察到。此外，抗 POSTN Ab 从 HPL 细胞中恢复了 MSC 共培养物中对成骨作用和 SOX11 mRNA 表达的抑制。然而，POSTN 诱导了 miR-299-5p 和 SOX11 的表达，并增强了成骨作用。
结论
HPL 细胞的体液因子抑制了 hMSCs 的成骨作用。这种抑制作用是通过 hMSCs 中的 miR-299-5p 和 SOX11 介导的。
Zotero 插件