Persistent Exposure to Fusobacterium nucleatum Triggers Chemokine/Cytokine Release and Inhibits the Proliferation and Osteogenic Differentiation Capabilities of Human Gingiva-Derived Mesenchymal Stem Cells

Abstract

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 Kang et al. Effects of F. nucleatum on GMSCs 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.

Time-Course Transcriptome Analysis of Gingiva-Derived Mesenchymal Stem Cells Reveals That Fusobacterium nucleatum Triggers Oncogene Expression in the Process of Cell Differentiation

“…The mRNA expression profile of GSE126821 in the GEO database 1 has been reanalyzed, which has been deposited according to our previous study (Kang et al, 2019). Briefly, three healthy individuals who underwent impacted tooth extraction at Stomatology Hospital of Shandong Province were recruited to participate in this study.…”

Modulation of the host cell transcriptome and epigenome by Fusobacterium nucleatum

“…Upregulation of CCL20 and CSF-3 (G-CSF) has been shown in Fn-infected esophageal tumor tissue and gingival fibroblasts, respectively (Kang et al, 2019a;Yamamura et al, 2016). CSF-2 (GM-CSF) upregulation has been observed in Fn-infected gingiva-derived mesenchymal stem cells (Kang et al, 2019b), and a role for CSF-2 in CRC epithelial-mesenchymal transition has been proposed (Chen et al, 2017). We show upregulation of CCL20, CSF-3, and CSF-2 is induced in both HCE and HCAE cells upon exposure to all Fn strains tested in our study.…”

Gingiva-Derived Mesenchymal Stem Cells: Potential Application in Tissue Engineering and Regenerative Medicine - A Comprehensive Review

“…Regarding the effect of bacteria on GMSCs, Kang W et al found that persistent exposure to F. nucleatum promotes cell migration and chemokine/cytokine release but suppresses the proliferation and osteogenic differentiation of GMSCs (148). In vivo, Su Y et al reported that altered oral microbiome and an increased LPS level caused by antibiotic treatment led to GMSCs deficiency and proliferation inhibition and a delayed wound healing in mice (122).…”