Reparative Dentin Formation Following Dental Pulp Capping is Mediated by TNFR1 In Vivo.

INTRODUCTION: Tumor necrosis factor (TNF)-α is a pro-inflammatory cytokine that promotes biomineralization in vitro in dental pulp cells. However, the role of TNF-α-TNF receptor 1 (TNFR1) signaling in reparative dentin formation and related inflammatory pathways is not known. Therefore, the aim of this study was to evaluate the role of the TNF-α-TNFR1 axis in dental pulp repair following pulp capping in vivo. METHODS: Dental pulp repair response of genetically deficient TNF-α receptor-1 mice (TNFR1-/-; n = 20) was compared with that of C57Bl6 mice (wild type [WT]; n = 20). Pulp capping was performed with mineral trioxide aggregate on the mandibular first molars of mice. After 7 and 70 days, tissues were collected and stained with hematoxylin and eosin for histopathological and histometric evaluation, and assessed by the Brown and Brenn methods for histomicrobiological analysis and by immunohistochemistry to localize TNF-α, Runt-related transcription factor 2, Dentin Sialoprotein (DSP) and Osteopontin (OPN) expression. RESULTS: Compared with WT mice, TNFR1-/- mice showed significantly decreased reparative dentin formation with a lower mineralized tissue area (P < .0001). Unlike WT mice, TNFR1-/- mice also exhibited significant dental pulp necrosis, neutrophil recruitment, and apical periodontitis formation (P < .0001) without bacterial tissue invasion. TNFR1-/- animals further exhibited decreased TNF-α, DSP, and OPN expression (P < .0001), whereas Runt-related transcription factor 2 expression was unchanged (P > .05). CONCLUSION: The TNF-α-TNFR1 axis is involved in reparative dentin formation following dental pulp capping in vivo. Genetic ablation of TNFR1 modified the inflammatory process and inhibited the expression of the DSP and OPN mineralization proteins, which culminated in dental pulp necrosis and development of apical periodontitis.

Could a chelant improve the effect of curcumin-mediated photodynamic antimicrobial chemotherapy against dental intact biofilms?

To our knowledge, there is still no evidence in relation to the combination of curcumin with chelants to improve the effects of antimicrobial photodynamic therapy (aPDT) on complex dental caries biofilms. Therefore, the aim of this study was to evaluate the antimicrobial effect of curcumin-ethylenediaminetetraacetic acid (EDTA)-mediated aPDT on the vitality of intact biofilms of dentin caries microcosms. Biofilms were grown on glass slabs in McBain medium plus 1% sucrose in microaerophily at 37 °C for 5 days. Then, biofilms were treated with associations of 600 µmol L-1 curcumin combined or not with 1% EDTA and 37.5 or 75 J cm-2 LED (455 nm). The vitality was determined by a confocal laser scanning microscopy (CLSM) after staining biofilms with a mixture of 2.5 g L-1 fluorescein diacetate and 0.25 g L-1 ethidium bromide. Statistical analysis was conducted by Kruskal-Wallis and post hoc Dunn's test (P < 0.05). Three treatments were able to reduce the vitality of overall biofilms: curcumin + 75 J cm-2 LED, curcumin-EDTA + 37.5 J cm-2 LED, and curcumin-EDTA + 75 J cm-2 LED. Also, the vitality of inner layers of biofilms was significantly reduced only after the combination of aPDT with EDTA. Therefore, the association of curcumin and EDTA improved the antimicrobial effect of aPDT on dentin caries microcosms, considering the application of lower light densities and deeper layers of biofilms.

Could chlorhexidine be an adequate positive control for antimicrobial photodynamic therapy in- in vitro studies?

BACKGROUND: Chlorhexidine digluconate (CHX) is commonly applied as positive control of new antimicrobials, because it is considered the gold-standard for chemical plaque control. The aim of this study was to compare the effect of treatments with curcumin-mediated aPDT and CHX in relation to the viability of specific microorganism groups in two distinct times (immediately and 24 h later). METHODS: Dentin caries microcosms were grown on bovine dentin discs (37 °C, anaerobiosis) for 3 days in the Active Attachment Amsterdam Biofilm Model. The biofilms were treated with 300 µM curcumin and 75 J.cm-² LED, or 0.06% and 0.12% CHX. Then, total microorganisms, total streptococci, mutans streptococci, and total lactobacilli counts were determined. The statistical analysis was conducted by Kruskal-Wallis and post-hoc Dunn's tests (P < 0.05). RESULTS: Curcumin-mediated aPDT (C + L+), 0.06% and 0.12% CHX reduced mutans streptococci counts (0.19, 0.10 and 0.07 log10 respectively) in the immediate analysis. After 24 h, it was observed a re-growth of microorganisms treated by curcumin-mediated aPDT, whereas both CHX concentrations demonstrated a decrease of the viable microorganisms. CONCLUSION: This study confirmed the substantive effect of CHX and the immediate effect of aPDT. The use of a neutralizer solution was important to block the substantivity of CHX and permit its fair comparison with aPDT, allowing its use as a positive control in further studies.