IADR Abstract Archives

Inflammatory Phenotype of Syncytiotrophoblasts Stimulated with Periodontal Bacteria and Hyperglycemia

Objectives: To assess a protocol for syncyalization of a BeWo cells.
To assess BeWo-SYN mRNA expression of TNF-α, IL-1β, TLR-2, TLR-4 and TLR-9 upon inactivated P. gingivalis and F. nucleatum in vitro stimulation.
To assess BeWo-SYN secretion of Adiponectin, IL-8, PLGF, CRP, IL-6, Leptin and TNF-α upon inactivated P. gingivalis and F. nucleatum and hyperglycemia in vitro stimulation.
Methods: BeWo cell line was syncytialized by the use of forskolin at different concentrations and time, evaluation of syncytialization was assessed by syncytialization ratio by immunofluorescence and expression of LGALS13 and PLGF mRNA by RT-qPCR. BeWo-SYN were stimulated with hyperglycemia, or heat-inactivated P.gingivalis and F.nucleatum (MOI 200 each and 100, both) for 15 min, 2, 24 and 48 h. mRNA expression of TNF-α, IL-1β, TLR-2, TLR-4 and TLR-9 quantified through RT-qPCR. Secreted adiponectin, IL-8, PLGF, CRP, IL-6, Leptin and TNF-α in conditioned medium at 24 and 48 hours was quantified by multiplex immunoassay. Comparisons of multiple variables at different time points was performed by ANOVA two-way.
Results: Syncytialization ratio of BeWo cells increased with time and concentration of forskolin with statistical differences (p<0.0001). LGALS13 and PLGF mRNA expression was higher at 48 h of 50 uM FSK. BeWo-SYN did not express TLR-2 mRNA. P.gingivalis MOI200 had a peak of TLR-4 and TLR-9 mRNA expression at 15 min, P.ginvialis/F.nucleatum MOI100 of TLR-9 at 2h, F.nucleatum MOI200 of TLR-4 and TLR-9 at 48 h. TNF-a mRNA expression was higher in control until 2 h, then it increased in F.nucleatum MOI200 at 24 h. Only IL-6 and Leptin were quantifiable inside the curve and leptin concentration had differences between bacterial stimulation groups (p-value=0.0324).
Conclusions: BeWo-SYN model of syncytiotrophoblast did not increase pro-inflammatory protein in CM upon bacterial and hyperglycemia stimuli, which is congruent with mRNA expression of pro-inflammatory markers. Bacterial translocation into placenta may provoke a proinflammatory stimuli, but BeWo SYN may have other hormonal responses, like reduced leptin secretion upon P.gingivalis stimulus.

2021 Chilean Division Annual Meeting (Virtual) 2021

2021

Periodontal Research-Pathogenesis
  • Bendek Viera, María José  ( Universidad de los Andes , Santiago , Chile )
  • Realini, Ornella  ( Universidad de los Andes , Santiago , Chile )
  • Monteiro, Lara  ( Universidad de los Andes , Santiago , Chile )
  • Acuña, Stephanie  ( Universidad de los Andes , Santiago , Chile )
  • Hernández, Marcela  ( Universidad de Chile , Santiago , Chile )
  • Hoare, Anilei  ( Universidad de Chile , Santiago , Chile )
  • Chaparro, Alejandra  ( Universidad de los Andes , Santiago , Chile )
  • none
    Fondecyt 1211471
    Oral Session
    Temas Libres 2