Fig 2: Innate immunity in the odontoblast layer (ODL). The effect of IL-1ß/IL1B, TNF-a/TNFA, IFN?/IFNG, and LPS on antimicrobial peptide regulation determined by real-time quantitative PCR (A). The effect of caries on HBD2 mRNA expression was detected by semi-quantitative PCR (B). IL-1ß, TNF-a, and LPS amplify HBD2 transcription in odontoblast-like cells in vitro. Consistently, caries increase expression of HBD2 mRNA in cells of ODL in vivo. Values are reported as relative fold change in mRNA transcription of unstimulated versus stimulated samples. The data represent means and standard errors from triplicate wells of one experiment and are representative of at least three independent experiments. Asterisks indicate statistically significant changes, with P < 0.05. A diagram illustrates the proposed caries-induced cellular interactions among cells in ODL through pro-inflammatory cytokines and chemokines (C). Bacterial components from caries activate cytokine/chemokine release from odontoblasts, dendritic cells, and/or macrophages via toll-like receptors (TLRs). Proinflammatory cytokines released from these cells act as autocrine and paracrine signals to amplify cytokine responses including antimicrobial peptide, cytokine, and chemokine production. The release of chemokines creates a migration gradient for immune cells to ODL while antimicrobial peptides reduce bacterial load.

Fig 3: Quantitative analysis for differential changes in the odontoblast layer (ODL) and pulp of carious teeth. Expression of inflammatory genes in ODL and pulp of normal versus carious teeth was determined by real-time quantitative PCR arrays (A-C). The fold change of gene expression detected by PCR arrays was more robust than that determined by cDNA arrays. However, the overall regulatory profile was similar: the increase of inflammatory gene expression in carious teeth was much more profound in ODL than in the pulp. Values are reported as relative fold change in mRNA transcription of carious versus normal samples. The data represent means and standard errors from triplicate sets of array analyses. Abbreviations: IFNA2, Interferon alpha 2; CEBPB, Enhancer binding protein beta; CRP, C-reactive protein; C4A, complement component 4A; BCL6, B-cell CLL/lymphoma 6; ABCF1, ATP-binding cassette, subfamily F member 1; CD40LG, CD40 ligand; TNF, Tumor necrosis factor alpha; SPP1, Secreted phosphoprotein 1 (osteopontin, bone sialoprotein I, early T-lymphocyte activation 1); LTB4R, Leukotriene B4 receptor; LTB, Lymphotoxin beta; LTA, Lymphotoxin alpha; TOLLIP, Toll-interacting protein; ICEBERG, ICEBERG caspase-1 inhibitor; IL, Interleukin; IL1R1, Interleukin 1 receptor, type I; IL1RN, Interleukin 1 receptor antagonist; IL10RA, interleukin 10 receptor alpha; IL10RB; interleukin 10 receptor beta; IL1F9, Interleukin 1 family member 9; XCR1, Chemokine (C motif) receptor 1; CX3CR1, Chemokine (C-X3-C motif) receptor 1; CCR, Chemokine (C-C motif) receptor; SCYE1, Small inducible cytokine subfamily E member1 (endothelial monocyte activating); MIF, Macrophage migration inhibitory factor; CXCL, Chemokine (C-X-C motif) ligand; CCL, Chemokine (C-C motif) ligand.