Dataset Information

Synergistic effects triggered by simultaneous Toll-like receptor-2 and -3 activation in human periodontal ligament stem cells.

ABSTRACT:

Background

Although periodontitis is associated with disruption of the host-microbial homeostasis, viruses are currently discussed to influence disease progression. Viral pathogens are recognized by Toll-like receptor (TLR)-3, which engages a different signaling pathway than other TLRs. This study aimed to investigate the effect of TLR-3 agonist polyinosinic:polycytidylic acid (Poly I:C) on the expression of inflammatory markers and bone metabolism proteins by human periodontal ligament stem cells (hPDLSCs) compared with TLR-2 agonist Pam3CSK4, which mimics the effect of bacterial lipoproteins. To assess potential combined effects of bacterial and viral infections, hPDLSCs response to simultaneous TLR-2 and TLR-3 activation was investigated.

Methods

HPDLSCs were stimulated with Poly I:C (0.0001-1 µg/mL), Pam3CSK4 (1 µg/mL), and their combinations for 24 hours. Gene expression and protein levels of interleukin (IL)-6, IL-8, monocyte chemoattractant protein (MCP)-1, and osteoprotegerin (OPG) were measured with qPCR and ELISA.

Results

Production of IL-6, IL-8, MCP-1, and OPG was significantly increased by Poly I:C or Pam3CSK4 to a similar extent. The levels of all inflammatory mediators induced by simultaneous stimulation with Poly I:C and Pam3CSK4 were significantly higher compared with single stimuli as well as to their summed response. Gene expression and protein levels of OPG were enhanced by Poly I:C, but by lesser extent than by Pam3CSK4. OPG levels upon simultaneous stimulation with Pam3CSK4 and Poly I:C were significantly lower compared with Pam3CSK4 stimulation alone.

Conclusions

Simultaneous TLR-2 and TLR-3 activation synergistically triggers IL-6, IL-8, and MCP-1 production, which was not observed for OPG. These findings suggest that TLR-3 activation by viral infections might promote periodontitis progression.

SUBMITTER: Blufstein A

PROVIDER: S-EPMC6852053 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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Publications

Synergistic effects triggered by simultaneous Toll-like receptor-2 and -3 activation in human periodontal ligament stem cells.

Blufstein Alice A Behm Christian C Gahn Johannes J Uitz Oksana O Naumovska Ivana I Moritz Andreas A Rausch-Fan Xiaohui X Andrukhov Oleh O

Journal of periodontology 20190529 10

<h4>Background</h4>Although periodontitis is associated with disruption of the host-microbial homeostasis, viruses are currently discussed to influence disease progression. Viral pathogens are recognized by Toll-like receptor (TLR)-3, which engages a different signaling pathway than other TLRs. This study aimed to investigate the effect of TLR-3 agonist polyinosinic:polycytidylic acid (Poly I:C) on the expression of inflammatory markers and bone metabolism proteins by human periodontal ligament ...[more]

PMID: 31049957

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Project description:BACKGROUND: Periodontal ligament mesenchymal stem cells (PDLSCs) are a promising cell resource for cell-based regenerative medicine in dentistry. PDLSCs inevitably acquire a senescent phenotype after prolonged in vitro expansion, and the key regulators of cells during replicative senescence remain unclear.METHODS: We cultured periodontal ligament stem cells to passages 4, 10 and 20 (P4, P10, P20). The senescent phenotypes, proliferation and migration ability of PDLSCs (P4, P10, P20) were detected, and non-targeted metabonomic sequencing was performed. We treated PDLSCs with AICAR and detected the expression of FOXO1, FOXO3a, FOXO6 and AMPK phosphorylation (p-AMPK) levels of replicating senescent PDLSCs to explore the correlation between the metabolic changes and the AMPK pathway.RESULTS: Immunofluorescence staining of γ-H2AX, β-galactosidase staining, cell scratch test and qPCR were performed to confirm the occurrence of replicative senescence in PDLSCs during passaging. Three groups of cells at passage 4 (P4), passage 10 (P10) and passage 20 (P20) were collected for non-targeted metabolomics analysis. Metabonomic sequencing showed that the metabolism of replicative senescence in PDLSCs varied significantly. In particular, the content of fatty acid metabolites decreased with senescence, including capric acid, stearic acid, myristic acid and dodecanoic acid. KEGG pathway analysis showed that the AMPK signaling pathway was closely related to AMP levels. The AMP:ATP ratio increased in senescent PDLSCs; however, the levels of p-AMPK and the profile of FOXO1 and FOXO3a, which are downstream of the AMPK signaling pathway, decreased with senescence. We treated PDLSCs with AICAR, an activator of the AMPK pathway, and the phosphorylated AMPK level at P20 PDLSCs was partially restored. CONCLUSION: In summary, our study suggests that the metabolic process of PDLSCs is active in the early stage of senescence, prefers to consume fatty acids, and is attenuated in the later stages of senescence. AMP accumulates in replicative senescent PDLSCs; however, the sensitivity of AMPK phosphorylation sites is impaired, causing senescent PDLSCs to fail to respond to changes in energy metabolism. Our findings provide a new basis for the clinical application of periodontal ligament stem cells.

Dataset Information

Synergistic effects triggered by simultaneous Toll-like receptor-2 and -3 activation in human periodontal ligament stem cells.

Background

Methods

Results

Conclusions

Publications

Synergistic effects triggered by simultaneous Toll-like receptor-2 and -3 activation in human periodontal ligament stem cells.

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