IADR Abstract Archives
HBMSC-Derived Exosomes Mitigate Th17/Treg Homeostasis in Periodontitis
Objectives: Exosomes are mediators of cell-to-cell communication in periodontitis. We aimed to investigate the role of hBMSCs-derived exosomes in regulating the balance of Th17/Treg in periodontitis.
Methods: The ratios of Th17/Treg in peripheral blood of periodontitis patients and healthy subjects were analyzed by flow cytometry. In vitro, exosomes were cocultured with human naïve CD4+ T cells exposed to pgLPS for 24h and the ratio of Th17/Treg was examined. The expression of inflammatory cytokines was determined by ELISA and qPCR. In vivo, HBMSCs-derived exosomes were administrated into mice periodontal pocket with periodontitis, and bone resorption was assessed by micro-CT. Furthermore, we pinpointed the regulatory miRNA in exosomes through miRNA sequencing and bioinformatics analysis. The interaction between miRNA and target proteins was verified using the luciferase activity assay. Lastly, the downstream Hippo pathway was detected by qPCR and western blotting.
Results: The ratio of Th17/Treg is out of balance in the peripheral blood of periodontitis patients. hBMSCs-derived exosomes decreased Th17/Treg ratio in CD4+ T cells in vitro and ameliorated inflammation and bone defects in periodontitis mice. Additionally, miR-1246 mimics induced HBMSCs-derived exosomes showed the enhanced effects of down-regulating the ratio of Th17/Treg in vitro. Furthermore, angiotensin converting enzyme2 (ACE2) was found to be the target protein in CD4+ T cells, and YAP-Hippo pathway was inhibited by miR-1246 from hBMSCs-derived exosomes.
Conclusions: hBMSCs-derived exosomes could alleviate periodontal inflammation through modulating the balance between Tregs and Th17 cells via targeting ACE2 and downregulating Hippo signaling pathway, which shed light on cell-free treatment for periodontal regeneration.
Methods: The ratios of Th17/Treg in peripheral blood of periodontitis patients and healthy subjects were analyzed by flow cytometry. In vitro, exosomes were cocultured with human naïve CD4+ T cells exposed to pgLPS for 24h and the ratio of Th17/Treg was examined. The expression of inflammatory cytokines was determined by ELISA and qPCR. In vivo, HBMSCs-derived exosomes were administrated into mice periodontal pocket with periodontitis, and bone resorption was assessed by micro-CT. Furthermore, we pinpointed the regulatory miRNA in exosomes through miRNA sequencing and bioinformatics analysis. The interaction between miRNA and target proteins was verified using the luciferase activity assay. Lastly, the downstream Hippo pathway was detected by qPCR and western blotting.
Results: The ratio of Th17/Treg is out of balance in the peripheral blood of periodontitis patients. hBMSCs-derived exosomes decreased Th17/Treg ratio in CD4+ T cells in vitro and ameliorated inflammation and bone defects in periodontitis mice. Additionally, miR-1246 mimics induced HBMSCs-derived exosomes showed the enhanced effects of down-regulating the ratio of Th17/Treg in vitro. Furthermore, angiotensin converting enzyme2 (ACE2) was found to be the target protein in CD4+ T cells, and YAP-Hippo pathway was inhibited by miR-1246 from hBMSCs-derived exosomes.
Conclusions: hBMSCs-derived exosomes could alleviate periodontal inflammation through modulating the balance between Tregs and Th17 cells via targeting ACE2 and downregulating Hippo signaling pathway, which shed light on cell-free treatment for periodontal regeneration.