What is the response profile of deciduous pulp fibroblasts stimulated with E. coli LPS and E. faecalis LTA?

Abstract

Background Oral fibroblast immunological responses to bacterial stimuli are well known. However, there are few studies about pulp fibroblasts from deciduous teeth (HDPF) responses, which are important for the treatment of pulp infections in children. The aim of this study was to evaluate expression and production of inflammatory cytokines and chemokines by HDPF when challenged with bacterial antigens normally present in advanced caries lesions. Methods Triplicate HDPF from 4 children (n = 4; 2 boys and 2 girls) were cultured by explant technique and challenged or not with Escherichia coli lipopolysaccharide/1 μg/mL (EcLPS) or Enterococcus faecalis lipoteichoic acid/1 μg/mL (EfLTA) for 6 and 24 h. Most of published studies employed immortalized cells, i.e., without checking possible gender and genetic variables. mRNA expression and protein production were evaluated by RT-qPCR and ELISA MILLIPLEX®, respectively, for Interleukin (IL)-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-17, Chemokine C-C motif ligand 2/monocyte chemoattractant protein 1 (CCL2/MCP-1), Chemokine C-C motif ligand 3/macrophage inflammatory protein 1-alpha (CCL3/MIP1-α), Chemokine C-C motif ligand 5/ regulated on activation, normal T cell expressed and secreted (CCL5/RANTES), C-X-C motif chemokine 12/ stromal cell-derived factor 1 (CXCL12/SDF-1), Tumor Necrosis Factor-alpha (TNF-α), Interferon-gamma (IFN γ), Vascular Endothelial Growth Factor (VEGF), Colony stimulating factor 1 (CSF-1) and Macrophage colony-stimulating factor (M-CSF). Results EcLPS increased IL-1α, IL-1β, IL-8, CCL2, CCL5, TNF-α and CSF-1 mRNA and protein levels while EfLTA was only able to positively regulate gene expression and protein production of IL-8. Conclusion The results of the present study confirmed our hypothesis, since pulp fibroblasts from deciduous teeth are capable of increasing gene expression and protein production after being stimulated with EcLPS and EfLTA.

Role of Lipopolysaccharide, Derived from Various Bacterial Species, in Pulpitis—A Systematic Review

“…E. coli LPS stimulates the production of pro-inflammatory factors: CCL2, IL-8 and IL-6 [ 41 ]. Less intense upregulation occurs for IL-4, GCFS, GM-CSF and CCL5, as well as for IL-1β, IL-10, IL12p70, IL-17A, TNF-α, INF-λ [ 41 ] and TNFβ [ 42 ]. The maturation and release of IL-1β is stimulated by a combination of LPS and ATP.…”

Overexpression of proinflammatory cytokines in dental pulp tissue and distinct bacterial microbiota in carious teeth of Mexican Individuals

“…Additionally, they found a similar expression of the regulatory cytokine IL10 between healthy dental pulp tissue and dental pulp tissue exposed to moderate caries; the present study also reports no difference in IL-10. Similarly, Colombini-Ishikiriama et al (Colombini-Ishikiriama et al, 2020) and Keller et al (Keller et al, 2010) observed a differential expression of IL1b, IL6, IL8, and TNFa in cultures of pulp fibroblasts of deciduous teeth stimulated with Escherichia colilipopolysaccharide (EcLPS) or Enterococcus faecalislipoteichoic acid (EfLTA). However, they also detected a differential expression of IL10.…”

“…Similarly, Colombini-Ishikiriama et al. ( Colombini-Ishikiriama et al., 2020 ) and Keller et al. ( Keller et al., 2010 ) observed a differential expression of IL1β, IL6, IL8, and TNFα in cultures of pulp fibroblasts of deciduous teeth stimulated with Escherichia coli -lipopolysaccharide (EcLPS) or Enterococcus faecalis -lipoteichoic acid (EfLTA).…”

Determination of the optimal concentration and duration of C5aR antagonist application in an inflammatory model of human dental pulp cells

“…Studies have confirmed that in cases of shallow caries, Gram‐positive bacteria account for a higher proportion, while in cases of deep caries and pulpitis, the proportion of Gram‐negative bacteria is higher [ 6 ]. Previous studies reported that both LTA and LPS were used as an inflammation model for dental pulp cells [ 7 , 8 , 9 , 10 ]. Our previous studies compared dental pulp cell inflammation models constructed using LTA and LPS, and the different treatment times with LTA and LPS (1, 2, 3, 5 and 7 days) [ 5 ].…”