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1.
Braz. oral res. (Online) ; 32: e103, 2018. tab, graf
Artículo en Inglés | LILACS | ID: biblio-974462

RESUMEN

Abstract The aim of this study is to evaluate the expression of cytokines in response to mineral trioxide aggregate (MTA) plus selenium in germ-free mice with experimental furcal perforation. The first left maxillary molar was opened, and the furcal area was perforated and treated with post-MTA-Se (experimental group). The same surgical intervention was performed for the maxillary right first molar, which was treated with MTA (control group). Fifteen mice were sacrificed 7, 14, and 21 days after furcal perforation, and periapical tissue samples were collected. The mRNA expression levels of the cytokines TGF-β, TNF-α, IFN-γ, HPRT, IL-10, IL-4, RANK, RANKL, IL-1, and IL-17 were assessed by using real-time polymerase chain reaction. In the experimental group, at 21-days post-MTA-Se sealing, the mRNA levels of TNF-α and IL-10 were upregulated compared with those in the control group (p < 0.05). Futher assessment revealed basal mRNA expression levels of IL-1α, IFN-γ, RANK, RANKL, IL-17A, IL-4, and TGF-β, over long experimental times, in both the experimental and control groups (p > 0.05). In conclusion, MTA+Se sealing favoured increased expression of IL-10 and TNF-α at later time points (day 21).


Asunto(s)
Animales , Masculino , Femenino , Óxidos/farmacología , Materiales de Obturación del Conducto Radicular/farmacología , Selenio/farmacología , Citocinas/análisis , Silicatos/farmacología , Defectos de Furcación/tratamiento farmacológico , Compuestos de Calcio/farmacología , Compuestos de Aluminio/farmacología , Cavidad Pulpar/lesiones , Tratamiento del Conducto Radicular/métodos , Factores de Tiempo , Reproducibilidad de los Resultados , Resultado del Tratamiento , Defectos de Furcación/inmunología , Cavidad Pulpar/efectos de los fármacos , Cavidad Pulpar/inmunología , Combinación de Medicamentos , Reacción en Cadena en Tiempo Real de la Polimerasa , Diente Molar/efectos de los fármacos , Diente Molar/lesiones
2.
J. appl. oral sci ; 25(1): 90-100, Jan.-Feb. 2017. tab, graf
Artículo en Inglés | LILACS, BBO | ID: biblio-841165

RESUMEN

Abstract IL-10 expressing regulatory B cells (B10) play a key role in immune system balance by limiting excessive inflammatory responses. Effects of toll-like receptor signaling and co-stimulatory molecules on B10 activity during innate and adaptive immune responses are not fully understood. Objective This study is to determine the effects of P. gingivalis LPS and CpG on B10 cell expansion and IL-10 competency in vitro. Material and Methods Spleen B cells were isolated from C57BL/6J mice with or without formalin-fixed P. gingivalis immunization. B cells were cultured for 48 hours under the following conditions: CD40L, CD40L+LPS, CD40L+CpG, and CD40L+LPS+CpG in the presence or absence of fixed P. gingivalis. Percentages of CD1dhiCD5+ B cells were measured by flow cytometry. IL-10 mRNA expression and secreted IL-10 were measured by real-time quantitative PCR and by ELISA respectively. Results P. gingivalis LPS plus CD40L significantly increased CD1dhiCD5+ B cell percentages and secreted IL-10 levels in both immunized and non-immunized mice B cells in the presence or absence of P. gingivalis, compared with control group. Secreted IL-10 levels were significantly increased in CD40L+LPS treated group compared with CD40L treatment group in the absence of P. gingivalis. CpG plus CD40L significantly decreased CD1dhiCD5+ B cell percentages, but greatly elevated secreted IL-10 levels in immunized and non-immunized mice B cells in the absence of P. gingivalis, compared with CD40L treatment group. Conclusions P. gingivalis LPS and CpG differentially enhance IL-10 secretion and expansion of mouse B10 cells during innate and adaptive immune responses.


Asunto(s)
Animales , Lipopolisacáridos/fisiología , Interleucina-10/inmunología , Porphyromonas gingivalis/fisiología , Ligando de CD40/fisiología , Receptor Toll-Like 9/agonistas , Receptor Toll-Like 4/agonistas , Linfocitos B Reguladores/inmunología , Bazo/citología , Factores de Tiempo , ARN Mensajero/análisis , Ensayo de Inmunoadsorción Enzimática , Distribución Aleatoria , Células Cultivadas , Interleucina-10/análisis , Interleucina-10 , Receptor Toll-Like 9/fisiología , Receptor Toll-Like 4/fisiología , Reacción en Cadena en Tiempo Real de la Polimerasa , Inmunidad Innata , Ratones Endogámicos C57BL
3.
Braz. oral res. (Online) ; 31: e63, 2017. graf
Artículo en Inglés | LILACS | ID: biblio-952122

RESUMEN

Abstract This study was conducted to investigate the roles of different Toll-like receptor (TLR) signaling in Porphyromonas gingivalis (P. gingivalis)-induced and ligature-induced experimental periodontal bone resorption in mice. Wild-type (WT), TLR2 knockout (KO), TLR4KO, and TLR2&4 KO mice with C57/BL6 background were divided into three groups: control, P. gingivalis infection, and ligation. Live P. gingivalis or silk ligatures were placed in the sulcus around maxillary second molars over a 2-week period. Images were captured by digital stereomicroscopy, and the bone resorption area was measured with ImageJ software. The protein expression level of gingival RANKL was measured by ELISA. The gingival mRNA levels of RANKL, IL-1β, TNF-α, and IL-10 were detected by RT-qPCR. The results showed that P. gingivalis induced significant periodontal bone resorption in WT mice and TLR2 KO mice but not in TLR4 KO mice or TLR2&4 KO mice. For all four types of mice, ligation induced significant bone loss compared with that in control groups, and this bone loss was significantly higher than that in the P. gingivalis infection group. RANKL protein expression was significantly increased in the ligation group compared with that in the control group for all four types of mice, and in the P. gingivalis infection group of WT, TLR2 KO, and TLR4 KO mice. Expression patterns of RANKL, IL-1β, TNF-α, and IL-10 mRNA were different in the P. gingivalis infection group and the ligation group in different types of mice. In summary, P. gingivalis-induced periodontal bone resorption is TLR4-dependent, whereas ligation-induced periodontal bone resorption is neither TLR2- nor TLR4-dependent.


Asunto(s)
Animales , Periodontitis/microbiología , Pérdida de Hueso Alveolar/etiología , Porphyromonas gingivalis/patogenicidad , Modelos Animales de Enfermedad , Receptor Toll-Like 2/fisiología , Receptor Toll-Like 2/genética , Receptor Toll-Like 4/fisiología , Factores de Tiempo , Ensayo de Inmunoadsorción Enzimática , Reproducibilidad de los Resultados , Pérdida de Hueso Alveolar/microbiología , Factor de Necrosis Tumoral alfa/metabolismo , Interleucina-10/metabolismo , Ratones Noqueados , Receptor Toll-Like 2/análisis , Receptor Toll-Like 4/análisis , Receptor Toll-Like 4/genética , Interleucina-1beta/metabolismo , Receptor Activador del Factor Nuclear kappa-B/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Ligadura , Metabolismo , Ratones Endogámicos C57BL
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