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Detection and function of lipopolysaccharide and its purified lipid A after treatment with auxiliary chemical substances and calcium hydroxide dressings used in root canal treatment.
Marinho, A C S; To, T T; Darveau, R P; Gomes, B P F A.
Afiliación
  • Marinho ACS; Department of Restorative Dentistry, Endodontics Division, Piracicaba Dental School, State University of Campinas (UNICAMP), Piracicaba, SP, Brazil.
  • To TT; Department of Periodontics, University of Washington School of Dentistry, Seattle, WA, USA.
  • Darveau RP; Department of Periodontics, University of Washington School of Dentistry, Seattle, WA, USA.
  • Gomes BPFA; Department of Restorative Dentistry, Endodontics Division, Piracicaba Dental School, State University of Campinas (UNICAMP), Piracicaba, SP, Brazil.
Int Endod J ; 51(10): 1118-1129, 2018 Oct.
Article en En | MEDLINE | ID: mdl-29505121
AIM: To investigate the influence of auxiliary chemical substances (ACSs) and calcium hydroxide [Ca(OH)2 ] dressings on lipopolysaccharides (LPS)/lipid A detection and its functional ability in activating Toll-like receptor 4 (TLR4). METHODOLOGY: Fusobacterium nucleatum pellets were exposed to antimicrobial agents as following: (i) ACS: 5.25%, 2.5% and 1% sodium hypochlorite solutions (NaOCl), 2% chlorhexidine (CHX) (gel and solution) and 17% ethylenediaminetetraacetic acid (EDTA); (ii) intracanal medicament: Ca(OH)2 paste for various periods (1 h, 24 h, 7 days, 14 days and 30 days); (iii) combination of substances: (a) 2.5% NaOCl (1 h), followed by 17% EDTA (3 min) and Ca(OH)2 (7 days); (b) 2% CHX (1 h), afterwards, 17% EDTA (3 min) followed by Ca(OH)2 (7 days). Saline solution was the control. Samples were submitted to LPS isolation and lipid A purification. Lipid A peaks were assessed by matrix-assisted laser desorption ionization time-of-flight mass spectrom (MALDI-TOF MS) whilst LPS bands by SDS-PAGE separation and silver staining. TLR4 activation determined LPS function activities. Statistical comparisons were carried out using one-way anova with Tukey-Kramer post-hoc tests at the 5% significance level. RESULTS: Matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of control lipid A demonstrated the ion cluster at mass/charge (m/z) 1882 and an intense band in SDS-PAGE followed by silver staining of control LPS. In parallel, LPS control induced a robust TLR4 activation when compared to ACS (P ≤ .001). 5.25% NaOCl treatment led to the absence of lipid A peaks and LPS bands, whilst no changes occurred to lipid A/LPS after treatment with others ACS. Concomitantly, 5.25% NaOCl-treated LPS did not activate TLR4 (P < .0001). As for Ca(OH)2 , lipid A was not detected by MALDI-TOF nor by gel electrophoresis within 24 h. LPS treated with Ca(OH)2 was a weak TLR4 activator (P < .0001). From 24 h onwards, no significant differences were found amongst the time periods tested (P > 0.05). The addition of Ca(OH)2 for 7 days to cells treated either with 2.5% NaOCl or 2% CHX led to the absence of lipid A peaks and LPS bands, leading to a lower activation of TLR4. CONCLUSION: 5.25% NaOCl and Ca(OH)2 dressings from 24 h onwards were able to induce both, loss of lipid A peaks and no detection of LPS bands, rendering a diminished immunostimulatory activity through TLR4.
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Texto completo: 1 Bases de datos: MEDLINE Métodos Terapéuticos y Terapias MTCI: Plantas_medicinales Asunto principal: Irrigantes del Conducto Radicular / Hidróxido de Calcio / Lipopolisacáridos / Fusobacterium nucleatum / Receptor Toll-Like 4 / Lípido A Tipo de estudio: Diagnostic_studies Idioma: En Revista: Int Endod J Año: 2018 Tipo del documento: Article País de afiliación: Brasil

Texto completo: 1 Bases de datos: MEDLINE Métodos Terapéuticos y Terapias MTCI: Plantas_medicinales Asunto principal: Irrigantes del Conducto Radicular / Hidróxido de Calcio / Lipopolisacáridos / Fusobacterium nucleatum / Receptor Toll-Like 4 / Lípido A Tipo de estudio: Diagnostic_studies Idioma: En Revista: Int Endod J Año: 2018 Tipo del documento: Article País de afiliación: Brasil