TLR2 agonist prevents the progression of periapical lesions in mice by reducing osteoclast activity and regulating the frequency of Tregs.

AIM: To evaluate the role of regulatory T lymphocytes (Tregs) in the presence or absence of the synthetic ligand Pam3Cys during the progression of periapical lesion in wild-type (WT) and toll-like receptor 2 knockout (TLR2KO) mice. METHODOLOGY: A total of 130 C57BL/6 male WT and TLR2KO mice were allocated into control (n = 5) and experimental (periapical lesion induction) (n = 10) groups. In specific groups (WT+Pam3cys and TLR2KO+Pam3cys), the synthetic ligand Pam3cys was administered intraperitoneally every 7 days, according to the experimental period (14, 21 and 42 days). At the end of those periods, the animals were euthanized, and the mandible and the spleen were submitted to histotechnical processing. Mandible histological sections were analysed by haematoxylin and eosin, TRAP histoenzymology and immunohistochemistry (FOXP3, RANK, RANKL and OPG). Spleen sections were analysed by immunohistochemistry (FOXP3). RESULTS: The inflammatory infiltrate and bone resorption were more intense in the TLR2KO group compared to the WT group. The animals that received the Pam3cys had smaller periapical lesions when compared to the animals that did not receive the ligand (p < .05). TLR2KO animals showed a significant increase in the number of osteoclasts when compared to TLR2KO+Pam3cys group (p < .05). At 21 days, the WT+Pam3cys group had a lower number of osteoclasts when compared to the WT animals (p = .02). FOXP3 expression was more intense in the WT+Pam3cys groups when compared to the WT animals in the 42 days (p = .03). In the spleen analysis, the WT+Pam3cys group also had a higher expression of FOXP3 when compared to the WT animals at 14 and 42 days (p = .02). Concerning RANKL, there was a reduction in staining in the KOTLR2+Pam3cys groups at 21 and 42 days (p = .03) and a higher binding ratio between RANK/RANKL in animals that did not receive the ligand. CONCLUSION: Administration of the Pam3cys increased the proliferation of Tregs, showed by FOXP3 expression and prevented the progression of the periapical lesion in WT mice. On the other hand, in the TLR2KO animals, Treg expression was lower with larger areas of periapical lesions. Finally, systemic administration of the Pam3cys in KO animals was able to limit the deleterious effects of the absence of the TLR2 receptor.

Genetic polymorphism in the tumour necrosis factor alpha gene (G-308A) is associated with persistent apical periodontitis in Brazilians.

AIM: To investigate if there was an association between genetic polymorphisms in tumour necrosis factor (TNF)-⍺ and its receptors TNFRSF1A and TNFRSF1B with persistent apical periodontitis (PAP) in Brazilian subjects. METHODOLOGY: Patients who had pulpal necrosis and apical periodontitis at the time of treatment, with at least 1-year of follow-up after non-surgical root canal treatment were recalled. Three hundred and seventy eight subjects were included, 150 subjects with signs/symptoms of PAP and 228 subjects with root canal-treated teeth exhibiting healthy perirradicular tissues (healed). Genomic DNA was extracted from saliva and used for TNF-⍺ (rs1800629), TNFRSF1A (rs1800693) and TNFRSF1B (rs1061622) genotyping by real-time PCR. Genotypes and alleles frequencies were evaluated by c2 or Fisher's exact tests and odds ratios were implemented (α = 5%). RESULTS: The genetic polymorphism in TNF-α (rs1800629) was associated as a protective factor for the development of PAP (p < .05), once subjects who presented at least one allele A (AA+AG X GG), had a higher chance to lesion repair (p < .05). The polymorphisms rs1800693 and rs1061622 in TNF receptors (TNFRSF1A and TNFRSF1B, respectively) were not associated with the development of PAP (p > .05). CONCLUSIONS: The observed results demonstrate that polymorphism in TNF-α but not in its receptors is associated with PAP.

Use of the Probiotic Bifidobacterium animalis subsp. lactis HN019 in Oral Diseases.

The oral cavity is one of the environments on the human body with the highest concentrations of microorganisms that coexist harmoniously and maintain homeostasis related to oral health. Several local factors can shift the microbiome to a pathogenic state of dysbiosis. Existing treatments for infections caused by changes in the oral cavity aim to control biofilm dysbiosis and restore microbial balance. Studies have used probiotics as treatments for oral diseases, due to their ability to reduce the pathogenicity of the microbiota and immunoinflammatory changes. This review investigates the role of the probiotic Bifidobacterium animalis subsp. lactis (B. lactis) HN019 in oral health, and its mechanism of action in pre-clinical and clinical studies. This probiotic strain is a lactic acid bacterium that is safe for human consumption. It mediates bacterial co-aggregation with pathogens and modulates the immune response. Studies using B. lactis HN019 in periodontitis and peri-implant mucositis have shown it to be a potential adjuvant treatment with beneficial microbiological and immunological effects. Studies evaluating its oral effects and mechanism of action show that this probiotic strain has the potential to be used in several dental applications because of its benefit to the host.

Absence of Tumor Necrosis Factor Receptor 1 Inhibits Osteoclast Activity in Apical Dental Resorption Caused by Endodontic Infection in Mice.

INTRODUCTION: The aim of this study was to evaluate osteoclastogenesis and dental resorption resulting from endodontic infection in wild-type (WT) and tumor necrosis factor receptor 1 genetically deficient (TNFR1 KO) mice. METHODS: After approval by the ethics committee on the use of animals, 40 mice were distributed into 2 experimental groups based on time periods: 14 days (n = 10 WT mice and n = 10 TNFR1 KO mice) and 42 days (n = 10 WT mice and n = 10 TNFR1 KO mice). After these periods, morphometric analysis was performed using bright field and fluorescence microscopy and tartrate-resistant acid phosphatase histoenzymology to identify osteoclasts. One-way analysis of variance followed by the Tukey post hoc test was used for the statistical analysis (α = 0.05). RESULTS: WT mice in the 42-day period had a greater apical dental resorption in the distal root of the first molar than TNFR1 KO mice (P < .05). On the other hand, TNFR1 KO mice showed a smaller number of osteoclasts on the dental surface than WT mice (P < .05). CONCLUSIONS: WT mice with apical periodontitis had more extensive apical dental resorptions and a larger number of osteoclasts on the tooth surface than TNFR1 KO mice.

Ex vivo evaluation of the effectiveness of XP-endo Finisher on the removal of smear layer from the root canal / Avaliação ex vivo da eficácia do XP-endo Finisher na remoção de smear layer do canal radicular

ABSTRACT Objective: The aim of this study was to evaluate the effectiveness of XP-endo Finisher (XP) on removal of the smear layer in root canals by comparing different irrigation protocols. Methods: Seventy-two human single-rooted teeth were similarly instrumented using R25 Reciproc files (VDW, Munich, Germany) applied in reciprocating mode with a VDW GOLD endo motor (VDW, Munich, Germany). The working length was determined at 1 mm short of the apical foramen. The canals were irrigated with 5 mL of 2.5% sodium hypochlorite during instrumentation. The teeth were divided at random into six groups (n=12). A control group, which was not submitted to the final irrigation protocol, and five experimental groups with different irrigants and agitation techniques: EDTA/File, EDTA/XP, EDTA/Passive Ultrasonic Irrigation (PUI), Distilled Water (DW)/XP, and DW/PUI). Smear layer removal quality scores were assessed in the apical, middle, and cervical thirds of the root canal based on images obtained by scanning electron microscopy. Data were analyzed using the Kruskal-Wallis test, followed by two-by-two comparisons with the Dunn test (α=5%). Results: EDTA/File, EDTA/PUI, and EDTA/XP groups demonstrated significantly lower scores than the other groups (P<0.05) in all thirds evaluated. No significant difference was observed between the groups in which distilled water was used and the control group in all thirds evaluated (P> 0.05). Conclusion: The XP-endo Finisher file did not increase the efficiency of EDTA in removal of the smear layer in root canals.

RESUMO Objective: Avaliar a eficácia do XP-endo Finisher (XP) na remoção da smear layer em canais radiculares, comparando diferentes protocolos de irrigação. Métodos: Setenta e dois dentes humanos unirradiculares foram similarmente instrumentados usando limas R25 (VDW, Munich, Germany) aplicadas no modo reciprocrantes em um motor endodôntico VDW GOLD (VDW, Munich, Germany). O comprimento de trabalho foi determinado a 1 mm aquém do forame apical. Os canais foram irrigados com 5 mL de hipoclorito de sódio 2,5% durante a instrumentação. Os dentes foram divididos aleatoriamente em seis grupos (n=12). Um grupo controle, que não foi submetido ao protocolo final de irrigação, e cinco grupos experimentais com diferentes irrigantes e técnicas de agitação: EDTA/Lima, EDTA/XP, EDTA/Irrigação Ultrassônica Passiva (IUP), Água destilada (AD)/XP, e AD/IUP). Os escores de qualidade de remoção da camada de smear layer foram avaliados nos terços apical, médio e cervical do canal radicular com base em imagens obtidas por microscopia eletrônica de varredura. Os dados foram analisados pelo teste de Kruskal-Wallis, seguido de comparações dois a dois pelo teste de Dunn (α = 5%). Resultados: Os grupos EDTA/lima, EDTA/PUI e EDTA/XP demonstraram escores significativamente menores que os outros grupos (P <0,05). Não foi observada diferença entre os grupos que utilizaram água destilada e o grupo controle em todos os terços avaliados (P> 0.05). Conclusão: A lima XP-endo Finisher não aumentou a eficiência do EDTA na remoção da smear layer em canais radiculares.

M1 and M2 macrophages phenotypes modulation after stimuli with materials used in endodontic treatment.

The aim of this study was to evaluate the M1 and M2 macrophage modulation after stimuli with different materials used during endodontic treatment. In bone marrow-derived macrophage cell culture, from males C57BL/6 wild-type (WT) mice, gene expression analysis of markers to M1 and M2 macrophages was performed by qRT-PCR (Cxcl10, CxCL9, iNOS, Arg1, Chil3, Retnla and MRC1) and cytokine quantification by Luminex® (GM-CSF, IL-10, IL-6, IL-1ß and TNF-α) after exposure to the five endodontic sealers: AH Plus, Sealapex Xpress, Endosequence BC Sealer, BioRoot RCS and a calcium hydroxide-based paste. For normal values, ANOVA test was used, followed by Tukey post-test. For non-normal values, the Kruskall-Wallis test was used. BioRootTM RCS and EndoSequence BC SealerTM stimulated the highest expression of markers for M1 macrophages, while calcium hydroxide-based paste stimulated the lowest expression of these gene markers. For M2 protein markers, BioRootTM RCS presented the highest stimulation while calcium hydroxide-based paste also presented the lowest stimulation. It was concluded that all the evaluated filling materials increased the genetic expression of pro- and anti-inflammatory markers: TNF-α and IL-10 respectively. The others proinflammatory mediators showed differences against the filling materials. However, this process did not induce the inflammatory response polarization, resulting in a hybrid macrophage.

M1 and M2 macrophages phenotypes modulation after stimuli with materials used in endodontic treatment

Abstract The aim of this study was to evaluate the M1 and M2 macrophage modulation after stimuli with different materials used during endodontic treatment. In bone marrow-derived macrophage cell culture, from males C57BL/6 wild-type (WT) mice, gene expression analysis of markers to M1 and M2 macrophages was performed by qRT-PCR (Cxcl10, CxCL9, iNOS, Arg1, Chil3, Retnla and MRC1) and cytokine quantification by Luminex® (GM-CSF, IL-10, IL-6, IL-1β and TNF-α) after exposure to the five endodontic sealers: AH Plus, Sealapex Xpress, Endosequence BC Sealer, BioRoot RCS and a calcium hydroxide-based paste. For normal values, ANOVA test was used, followed by Tukey post-test. For non-normal values, the Kruskall-Wallis test was used. BioRootTM RCS and EndoSequence BC SealerTM stimulated the highest expression of markers for M1 macrophages, while calcium hydroxide-based paste stimulated the lowest expression of these gene markers. For M2 protein markers, BioRootTM RCS presented the highest stimulation while calcium hydroxide-based paste also presented the lowest stimulation. It was concluded that all the evaluated filling materials increased the genetic expression of pro- and anti-inflammatory markers: TNF-α and IL-10 respectively. The others proinflammatory mediators showed differences against the filling materials. However, this process did not induce the inflammatory response polarization, resulting in a hybrid macrophage.

Resumo O objetivo deste estudo foi avaliar a modulação dos macrófagos M1 e M2 após estímulos com diferentes materiais utilizados durante o tratamento endodôntico. Em cultura de células de macrófagos derivados da medula óssea de camundongos machos C57BL/6 wild-type (WT), após a exposição à cinco cimentos endodônticos: AH Plus, Sealapex Xpress, Endosequence BC Sealer, BioRoot RCS e pasta à base de hidróxido de cálcio foi realizada a análise da expressão gênica dos marcadores para macrófagos M1 e M2 por qRT-PCR (Cxcl10, CxCL9, iNOS, Arg1, Chil3, Retnla e MRC1) e quantificação de citocinas por Luminex® (GM -CSF, IL-10, IL-6, IL-1β e TNF-α). Para valores normais, foi utilizado o teste ANOVA, seguido do pós-teste de Tukey. Para valores não normais, foi utilizado o teste de Kruskall-Wallis. BioRootTM RCS e EndoSequence BC SealerTM estimularam maior expressão de marcadores para macrófagos M1, enquanto a pasta à base de hidróxido de cálcio estimulou expressão mais baixa desses marcadores gênicos. Para o marcador de proteínas para M2, BioRootTM RCS apresentou a maior estimulação, enquanto a pasta à base de hidróxido de cálcio também apresentou menor estimulação. Concluiu-se que os materiais obturadores avaliados aumentaram a expressão genética de marcadores pró- e anti-inflamatórios: TNF-α e IL-10 respectivamente. Os demais marcadores pró inflamatórios mostraram diferenças em relação aos materiais obturadores. No entanto, esse processo não induziu a polarização da resposta inflamatória, resultando em um macrófago híbrido.

Alteration of the oral microbiota may be a responsible factor, along with estrogen deficiency, by the development of larger periapical lesions.

OBJECTIVES: To answer the questions: (1) Does reducing estrogen levels influence the microbial composition of the oral cavity? (2) Does the presence of periapical lesion (PL) cause changes in the oral microbiota? (3) Since estrogen deficiency alters the oral microbiota, can this be one of the factors that contribute to the increase of the PL? MATERIALS AND METHODS: Thirty-six rats were divided into four groups: sham (control), ovariectomy (OVX), control with PL (Sham + PL), and OVX + PL. After 9 weeks of OVX, the lower first molars were submitted to PL induction. After 21 days, the microbiological collection of the oral cavity was performed, and the animals were euthanized. The contents were evaluated by the checkerboard DNA-DNA hybridization method, to verify the prevalence of 40 bacterial species (divided into 7 microbial complexes). The blocks containing the lower first molars were submitted to histotechnical processing and staining with hematoxylin and eosin (HE), for the measurement of the periapical lesion area. The results were submitted to ANOVA and Kruskal-Wallis tests and Tukey and Dunn post-tests, with a significance level of 5%. RESULTS: In conditions of estrogen deficiency, there was alteration of the oral microbiota. The OVX groups had a higher amount of bacteria compared to the SHAM group in most of the microbial complexes (p < 0.001). The animals in the control group (with or without lesion) did not present a statistically significant difference (p > 0.001) in any of the microbial complexes. The PLs in OVX animals were significantly higher compared to SHAM animals (p < 0.001). CONCLUSIONS: Hypoestrogenicity conditions interfere in the oral microbiota by increasing the amount of bacteria in the saliva and influencing the progression of periapical lesions. CLINICAL RELEVANCE: This inedited study shows that deficiency of estrogen leads to alteration of the oral microbiota.

Immunohistochemical characterization of immune cell infiltration in paediatric and adult Langerhans cell histiocytosis.

Langerhans cell histiocytosis (LCH) is an inflammatory myeloid neoplasia commonly affecting children with frequent somatic mutations in MAPK pathway genes including BRAFV600E and MAP2K1. Some studies suggest that LCH cells can recruit and modulate inflammatory cells, which could provide reciprocal survival signals. To characterize the immune profile of infiltrating inflammatory cells, and to clarify their participation in LCH pathogenesis, a detailed immunohistochemical analysis was performed. Fifteen (10 children, 5 adults) LCH cases were assessed through macrophage (CD68 and CD163), mature dendritic cell (mDC; CD83 and CD208), regulatory T cell (Treg; CD4, CD25 and FOXP3) and cytotoxic lymphocyte (CL; CD56, CD57, perforin and granzyme B) immunomarkers. Moreover, lymphocytic and LCH markers were also analysed. All cases were S100, CD1a, CD207 and CD4-positive. Bcl-2 and cyclin D1 expression was observed in 13 of 15 cases. In the immune microenvironment, M2-polarized macrophages and Tregs were the predominant cell populations, followed by significantly (P < .005) smaller levels of mDCs and CLs. Additionally, the number of CD3 + cells was significantly higher than that of CD20 + cells. In the CD3 + cell population, there were a significantly higher number of CD4 + cells than CD8 + cells. While there were no differences when comparing the paediatric and adult populations, FOXP3 + cells were significantly higher in patients with multisystem involvement and treated with chemotherapy, than single-site cases and those without chemotherapy. Our results suggest that M2-polarized macrophages and Treg infiltration can promote LCH development and survival, probably through pro-tumoral, immunosuppressive and/or cytokine-mediated mechanisms. This work highlights the need for further exploration of immune-targeted therapy for LCH.

Clinical, microbiological, and immunological evaluation of patients in corrective orthodontic treatment.

BACKGROUND: The objective was to analyze clinical, microbiological, and immunological periodontal parameters in patients in corrective orthodontic treatment. MATERIALS AND METHODS: Twenty-eight patients were selected. Plaque index (PI), bleeding on probing (BOP), width of keratinized gingiva, levels of 40 bacterial species, and of 3 cytokines (IL-1ß, MMP-8, and TNF-α) in gingival crevicular fluid (GCF) were evaluated at T0, before orthodontic treatment; T1, 6 months; and T2, 12 months post-treatment. Non-parametric, Friedman, Wilcoxon, ANOVA, and Spearman correlation coefficient tests were used for statistical analyses, with the significance level of 5%. RESULTS: No significant difference was found for the width of keratinized gingiva, but PI presented a significant increase at T1 and T2 (p < 0.05) when compared with T0. The percentage of sites with BOP increased significantly from T0 to T1 (p < 0.05); however, at T2, the values decreased and did not differ anymore from T0 (p > 0.05). In the microbiological analysis, red complex pathogens were in significantly greater proportions in T2 compared with T0 (p < 0.05). There was no statistically significant difference in the cytokine levels between the periods but there was a positive correlation between BOP and IL-1ß (r = 0.49 p = .01) and TNF-α (r = 0.39 and p = .05). CONCLUSION: In conclusion, corrective orthodontic treatment caused clinical periodontal alterations regarding biofilm accumulation and gingival bleeding, with alteration of periodontopathogens.

Effects of 5-lipoxygenase gene disruption on inflammation, osteoclastogenesis and bone resorption in polymicrobial apical periodontitis.

OBJECTIVES: To investigate the regulation of inflammatory and osteoclastogenic signaling by 5-lipoxygenase (5-LO) in apical periodontitis induced by oral contamination of dental root canals in mice. DESIGN: Apical periodontitis was induced in 5-lipoxygenase enzyme knockout (129-Alox5tm1Fun) and 129 wild-type mice (n = 96) by exposure of the dental root canal to the oral cavity. After 7, 14, 21, and 28 days, the animals were euthanized and the tissues removed (n = 12 teeth per period) for histopathological and histometric analyses (hematoxylin and eosin [HE]), evaluation of osteoclastogenic activity (tartrate-resistant acid phosphatase enzyme [TRAP]), and determination of inflammatory and osteoclastogenic signaling (qRT-PCR). RESULTS: Oral contamination of dental root canals induced recruitment of neutrophils and osteoclasts to the periodontal ligament, resulting in bone resorption. Absence of 5-LO did not impair neutrophil recruitment while osteoclastic formation was increased. Nonetheless, early bone resorption progressed similarly to lesions in wild-type animals. Interestingly, in the absence of 5-LO, the synthesis of mRNAs for cytokines, chemokines, and their receptors was significantly reduced while that of regulators of osteoclastogenesis (RANK, RANKL, and OPG) was increased in comparison with the corresponding levels in wild-type animals. CONCLUSIONS: The 5-LO pathway plays a role in the stimulation of inflammatory mediator synthesis and inhibition of osteoclastogenesis in apical periodontitis in mice. However, the paradoxical inflammatory-osteoclastogenic signaling did not impair inflammatory cell recruitment and bone resorption during early development of the disease.

Radiotherapy impairs adhesive bonding in permanent teeth.

OBJECTIVES: To evaluate the in vitro effects of radiotherapy (RT) on the morphological surface of the enamel and dentin and to determine the best adhesive system and most appropriate time to restore teeth in head and neck cancer patients. METHODS: Sixty third molars were cut into 120 enamel fragments and 120 dentin fragments and divided into four groups (n = 30): G1 (control): nonirradiated, only restorative procedure; G2: restorative procedure immediately before RT; G3: restorative procedure immediately after RT; and G4: restorative procedure 6 months after RT. Each group was divided into two subgroups: Adper™ Single Bond 2 (SB) and Clearfill SE Bond (CL) based on the material used. After RT and restorative procedures, the specimens were subjected to confocal microscopy and shear bond strength test. Data were analyzed using a two-way ANOVA followed by Tukey's test at a significance level of 5%. RESULTS: Morphological changes were observed in both substrates after a cumulative dose of 40 Gy, and after 60 Gy, the changes were more evident in both substrates. CL had the highest strength values in both substrates (p < 0.05), and G2 had the lowest strength values for the enamel and dentin (p < 0.05). CONCLUSIONS: Based on the in vitro study results, we can conclude that RT substantially changes the morphological surface of enamel and dentin and impairs the bond strength. The Clearfill system yielded better results than Adper Single Bond 2, and restoring teeth before RT resulted in the worst results in both substrates.

Examination of OPG, RANK, RANKL and HIF1A polymorphisms in temporomandibular joint ankylosis patients.

PURPOSE: To evaluate the association between polymorphisms in genes that regulate bone metabolism, such as OPG, RANK, RANKL, and HIF1A, in patients with temporomandibular joint (TMJ) ankylosis. METHODS: The sample consisted of 181 individuals, the study included 17 individuals with TMJ ankylosis and 164 controls. DNA was extracted from buccal epithelial cells. The genotyping of genetic polymorphisms in OPG (rs2073618), RANK (rs3826620), RANKL (rs9594738), and HIF1A (rs2301113 and rs2057482) was performed by real-time PCR using TaqMan™ technology (Applied Biosystems). The data were subjected to statistical analysis with a level of significance of 0.05. RESULTS: The OPG (rs2073618) polymorphism was associated with TMJ ankylosis, both in the additive model and in the dominant model (p < 0.05). In the additive model, when the individuals carried the CC genotype, they presented as 10.80 times more likely to develop the condition (p = 0.03). In the dominant model, individuals that carried at least one C allele were 5.76 times more likely to have TMJ ankylosis, than those with the G allele (p = 0.01). CONCLUSION: The polymorphism rs2073618 of OPG is a possible marker that is associated with the risk of manifestation of TMJ ankylosis.

RANKL is associated with persistent primary teeth and delayed permanent tooth emergence.

BACKGROUND: Tooth eruption is a process that is not fully understood. AIM: To evaluate whether genetic polymorphisms for RANK/RANKL/OPG are associated with delayed tooth emergence. To evaluate whether the relative expression of this genes is associated with persistent primary teeth. DESIGN: To evaluate whether genetic polymorphisms for RANK/RANKL/OPG could be involved in delayed tooth emergence, saliva samples from 160 children, aged 6-13 years old, were analysed. To test if there is correlation between gene expression of RANK/RANKL/OPG in children with delayed tooth emergence and persistent primary teeth, periapical tissue from 15 children with persistent primary teeth and from 15 control subjects were collected for qPCR analysis. RESULTS: Fifty-six children with delayed tooth emergence (35%) had at least one permanent tooth with delayed emergence. The T allele in RANKL (rs9594738) increased the risk of delayed tooth emergence (P = 0.02; OR = 1.71, 95%CI 1.09-2.75). The relative gene expression for RANKL and the ratio RANKL/OPG in children with delayed tooth emergence and persistent primary teeth were lower compared to controls (P = 0.02 and P = 0.005, respectively). CONCLUSIONS: Data suggest that the polymorphism rs9594738 in RANKL is associated with delayed permanent tooth emergence. Moreover, reduced relative gene expression of RANKL in periapical tissue is associated with persistent primary teeth.

Novel endodontic sealers induced satisfactory tissue response in mice.

OBJECTIVES: The aim of this study was to evaluate the subcutaneous response induced by Roeko Guttaflow2 (RG), Sealapex Xpress (SX), AH Plus (AHP) sealers. METHODS: 100 BALB/c mice received implants in the subcutaneous tissue with the tested materials (10 animals per period for each evaluated sealer) and were evaluated after different experimental periods (7, 21 and 63 days), in each animal was placed a tube, the control group was an empty tube. Histological analysis evaluated semi-quantitatively the inflammatory infiltration, collagen fiber formation and tissue thickness. In addition, immunohistochemistry was performed for interleukin-6 (IL-6). Data were statistically analyzed (α = 0.05). RESULTS: RG promoted a greater collagen fiber formation at 7 days and 63 days compared to the CG (p = 0.004) and AHP (p = 0.005) respectively, while at 21 days, the SX promoted a greater reaction (p = 0.021). For the tissue thickness, there was a greater reaction at 7 days with CG (p = 0.0156) and with RG at 63 days (p = 0.03). Regarding the inflammatory infiltrate, there was no difference at 7 days and 63 days (p = 0.5; p = 0.27), while at 21 days, a statistically difference was found between SX, CG (p = 0.04) and RG (p = 0.027). In addition, the presence of IL-6 was observed in almost all groups, with a more intense marking at 7days. SIGNIFICANCE: All cements evaluated presented a satisfactory tissue response, however, RG was the one that presented a more satisfactory tissue response.

The effect of ovariectomy and 2 antiresorptive therapeutic agents on bone response in rats: A 3-dimensional imaging analysis.

OBJECTIVE: The aim of this study was to evaluate bone mineral density (BMD) and microarchitecture in femurs and maxillary bones of ovariectomized (OVX) rats treated or not treated with alendronate (ALD) or odanacatib (ODN). STUDY DESIGN: Twenty rats were divided into groups: SHAM, OVX, OVX/ALD, and OVX/ODN. After 12 weeks, the femurs and maxillae were removed and subjected to 3-dimensional analysis by micro-computed tomography. Results were analyzed with 1-way analysis of variance and Tukey's post hoc test (α = 0.05). RESULTS: OVX decreased maxillary and femoral BMD and altered femoral microarchitecture (P < .05). The drugs increased BMD of both types of bones, but only ALD maintained the phenotype similar to the SHAM group. The action of ALD was limited to the femoral trabecular separation (Tb.Sp). OVX and the drugs had no effect on the microarchitecture of the maxilla (P > .05). CONCLUSIONS: ALD and ODN therapy increased BMD in both bones after ovariectomy. ALD was more successful than ODN in preserving the morphology of bone similar to the SHAM group. ALD maintained the phenotype for Tb.Sp in the femur, but ODN did not. In the maxillae, neither ovariectomy nor the 2 antiresorptive drugs had significant effects on microarchitecture.

Genetic polymorphism in RANK is associated with mandibular size.

OBJECTIVE: The aim of this study was to evaluate the association between genetic polymorphisms in RANK, RANKL and OPG with maxillary and mandibular dimensions in humans. DESIGN: DNA extracted from saliva and the rs3826620, rs9594738 and rs2073618 polymorphisms in RANK, RANKL and OPG, respectively, were analysed by real-time PCR. Four linear measurements (Co-Gn, GoPg, Co-Go and PTM-A) from lateral cephalograms were examined for the evaluation of craniofacial measurements. ANOVA testing and a multivariate linear regression analysis, adjusted for age and gender, were used for statistical analysis, with an alpha of 5%. Hardy-Weinberg equilibrium was also evaluated using the chi-square test within each polymorphism. SETTING: School of Dentistry of Ribeirão Preto, University of São Paulo. PARTICIPANTS: A total of 100 unrelated non-syndromic Brazilian Caucasian subjects were included in this study. RESULTS: The polymorphism in RANK was associated with a higher Go-Pg measurement (p = .039). In the multivariate analysis, adjusted for age and gender, the polymorphism in RANK was associated with Go-Pg (p = .017) and Co-Gn (p = .043). CONCLUSION: The polymorphism rs3826620 in RANK is associated with the mandibular size.

Cytokine profile changes in gingival crevicular fluid after placement different brackets types.

OBJECTIVE: The aim of this study was to examine the relationship between bracket design and ratio of five proinflammatory cytokine, in gingival crevicular fluid (GCF), and bacterial adhesion without tooth movement influence. DESIGN: The sample was comprised of 20 participants, aged 11 to 15 years old (mean age: 13.3 years±1.03). A conventional Gemini™ metallic bracket and two self-ligating brackets, In-Ovation®R and SmartClip™, were bonded to the maxillary incisors and canines. GCF was collected using a standard filter paper strip before and 60days after bonding. The cytokine levels (IL-12, IL-1α, IL-1ß, IL-6 and TNF-α) were performed by the LUMINEX assay. The levels of the red and orange bacterial complexes were analyzed by the Checkerboard DNA-DNA hybridization. The data of cytokine and bacterial complexes were carried out using the non-parametric tests at 5% of significance level. RESULTS: Increased cytokine levels were observed. However, only the SmartClip™ group showed a significantly increased level of TNF-α (p=0.046). The SmartClip™ brackets group presented higher levels of red complex bacteria. CONCLUSIONS: The bracket design affected cytokine levels and bacterial adhesion since it was observed that the proinflammatory cytokines released in GCF to the SmartClip™ group showed an increase in the TNF-α levels associated with higher bacterial levels, which possibly represents greater inflammatory potential. Thereby, the bracket design should be considered in patients with risk of periodontal disease and root resorption.

Cementocytes Express Receptor Activator of the Nuclear Factor Kappa-B Ligand in Response to Endodontic Infection in Mice.

INTRODUCTION: Although studies have recently shown that osteocytes embedded in mineralized bone matrix play an important role in bone diseases, the participation of cementocytes in apical periodontitis has not been evaluated. The aim of the present study was to evaluate the possible involvement of cementocytes in the development of apical periodontitis. METHODS: Apical periodontitis was experimentally induced in the lower first molars of wild-type mice by pulp exposure to the oral environment. At 0, 7, 21, and 42 days after pulp infection, the animals were euthanized, and the jaws were prepared for analysis under conventional and fluorescence microscopy (morphologic and morphometric analysis), immunohistochemistry and immunofluorescence (receptor activator of nuclear factor kappa-B [RANK], receptor activator of the nuclear factor kappa-B ligand [RANKL], and osteoprotegerin [OPG]), enzyme histochemistry (osteoclasts and cementoclasts), and real-time polymerase chain reaction (RANK, RANKL, OPG, and cathepsin K). RESULTS: At 7, 21, and 42 days after pulp exposure, there was a progressive increase in periodontal ligament, cementum and bone resorption areas, osteoclasts, and cementoclast counts as well as higher messenger RNA levels of RANK, RANKL, OPG, and cathepsin K. In intact teeth, cementocytes and osteocytes did not express RANKL. After infection, RANKL was strongly expressed in cementocytes, but not in osteocytes, and its expression increased with lesion progression. CONCLUSIONS: Our findings show that cementocytes express RANKL in response to endodontic infection and may be involved in the pathogenesis of apical periodontitis.

Lymphangioma of the Lower Lip Mimicking a Mucocele in Children.

Lymphangioma is a hamartoma resulting from the proliferation of common lymphatic vessels in the head and neck area, rarely appearing in the lower lips. Its clinical presentation is a nodular mass with a pebbly surface and no defined borders formed by a cluster of slow-growing coalescing vesicles. The purpose of this paper is to present two children with a lesion in the lower lip whose clinical characteristics (single papillary lesion with a pediculated base, same color and consistency of the surrounding mucosa, and negative result for diascopy test) suggested an initial diagnosis of mucocele but were later confirmed as cavernous lymphangiomas. The clinical and microscopic characteristics of the lesions are discussed as well as the differential diagnosis and the treatment approach. These cases reinforce the importance of always performing a confirmatory histopathological analysis, even for lesions with typical clinical features.