Oral conditions are associated with salt taste disability among American adults.

AIM: To explore the association between oral conditions and their interaction with salt taste disability among American adults. METHODS: Data from the 2013-2014 NHANES cycle were used (n = 2373). The exposures were periodontitis, defined by the 2017 EFP-AAP classification, dental caries, missing teeth, and edentulism, as per the DMF-T index, and xerostomia. The outcome was salt taste disability, objectively assessed. Covariates included sex, age, educational level, poverty index, obesity, diabetes, smoking, alcohol consumption, and medications related to mouth dryness. Weighted multivariable logistic regression modeling was used to evaluate the relationship between oral conditions and their interaction and salt taste disability. RESULTS: Participants who reported xerostomia were more likely to have salt taste disability (OR 2.42; 95%CI 1.44-4.07), especially those older than 60 years (OR 3.63; 95%CI 1.72-7.63). Among participants aged 40-59, xerostomia increased the chance of salt taste disability; however, the confidence interval included the null value. The interactions between xerostomia and edentulism increased the chance of salt taste disability. CONCLUSION: Oral conditions seem to influence the ability to taste salt. Dental professionals may help identify individuals with taste alterations and raise their awareness of the risk of systemic diseases that require the reduction of salt intake.

Incidence, mortality and survival rates of lip, oral cavity and salivary glands cancers in Singapore: A half-century time trend analysis (1968-2017).

OBJECTIVES: To examine trends in incidence and mortality and evaluate overall survival (OS) of oral cancer in Singapore between 1968 and 2017. METHODS: All diagnosed oral cancers by anatomical sites and population size were extracted from the Singapore Cancer Registry and the Department of Statistics Singapore. The trend of age-standardized incidence rate (ASIR) and mortality rate (ASMR) (per 100 000 person-years) of the lip, oral cavity and salivary gland cancers were evaluated by Prais-Winsten regressions for each ethnicity and gender. Kaplan-Meier curves were performed to evaluate the OS by anatomical sites in each age group by ethnicity and sex. RESULTS: Overall, 49, 3494 and 1066 people were diagnosed, and 28, 2310 and 476 died from lip, oral cavity and salivary gland cancers, respectively. The oral cavity cancer ASIR and ASMR reduced from 3.07 (1968-1972) to 2.01(2008-2012) and from 2.06 (1978-1982) to 1.21 (2013-2017) per 100 000 person-years, respectively, with both highest in Indians throughout the whole period. Male:Female ratio ranged from 3.43 (1973-1977) to 1.75 (2013-2017) and from 3.41 (1978-1982) to 2.40 (2013-2017) for ASIR and ASMR, respectively. However, both salivary gland cancer ASIR and ASMR increased from 0.50 (1968-1972) to 0.80 (2008-2012) and from 0.18 (1968-1982) to 0.42 (1988-1992) per 100 000 person-years, respectively, with both higher in males since 1993. Oral cavity cancer ASIR decreased for males aged ≥60, and Indian females ≥25, but increased among Chinese females aged ≥60. Oral cavity cancer ASMR decreased among Chinese aged 25-59, and among Malay males and Indian females. Salivary gland cancer ASIR increased among Chinese males aged ≥60 and Malay males aged 25-59; while ASMR increased among Chinese males aged ≥60. The median OS for oral cavity, lip and salivary gland cancers were 3.0, 9.3 and 18.1 years, respectively, with females surviving longer than males. CONCLUSIONS: Singapore has experienced a decline in the incidence and mortality of lip, oral cancer, an increase in in the incidence and mortality of salivary gland cancer, with an increase in the median overall survival rate. Monitoring the magnitude of oral cancer burden and the demographic, and temporal variations is necessary for tailoring health planning and setting priorities for future clinical care and research.

Chewing Disability Is Associated With Cognitive Impairment Among Older Adults: A Population-Based Cohort Study.

BACKGROUND: Chewing disability is associated with impaired quality of life, potentially leading to depression, and cognitive impairment. Although the chewing-ability-cognition relationship has been explored, examining whether depression mediates this relationship remains unclear. We investigated the association between chewing disability and cognitive impairment development and a potential mediation via depression among older persons. METHODS: Older persons without cognitive impairment at baseline (n = 973) from the 3 waves of the Panel on Health and Ageing of Singaporean Elderly were investigated. The outcome was incident cognitive impairment by the end of the study, while the exposure was chewing disability over the study period. Time-varying depression was the mediator. Time-fixed confounders included sex, ethnicity, education, marital status, living arrangement, and housing type, and time-varying confounders included age, smoking, cardiovascular diseases, diabetes, number of teeth, and denture wearing. We used marginal structural modeling to evaluate the effect of chewing disability on cognitive impairment development. RESULTS: After 6 years, 11% developed cognitive impairment, and chewing disability was reported by 33%. Chewing disability was associated with higher odds of developing cognitive impairment (OR 1.43, 95% CI: 1.09, 1.87), of which 85.3% was explained by the controlled direct effect of chewing disability, whereas the remaining 14.7% could be eliminated if there was no depression. CONCLUSIONS: Our findings indicate an association between chewing disability and cognitive impairment, while the role of depression could not be fully elucidated. Oral health should be incorporated as part of older persons' care for its potential to assess the risk for other systemic conditions.

Intake and serum levels of micronutrients and chronic oral diseases burden.

OBJECTIVE: To investigate pathways from micronutrient intake and serum levels to Chronic Oral Diseases Burden. METHODS: We analyzed cross-sectional data from NHANES III (n = 7936) and NHANES 2011-2014 (n = 4929). The exposure was the intake and serum levels of vitamin D, calcium, and phosphorus. Considering the high correlation of those micronutrients in the diet, they were analyzed as a latent variable dubbed Micronutrient intake. The outcome was the Chronic Oral Diseases Burden, a latent variable formed by probing pocket depth, clinical attachment loss, furcation involvement, caries, and missing teeth. Pathways triggered by gender, age, socioeconomic status, obesity, smoking, and alcohol were also estimated using structural equation modeling. RESULTS: In both NHANES cycles, micronutrient intake (p-value < 0.05) and vitamin D serum (p-value < 0.05) were associated with a lower Chronic Oral Diseases Burden. Micronutrient intake reduced the Chronic Oral Diseases Burden via vitamin D serum (p-value < 0.05). Obesity increased the Chronic Oral Diseases Burden by reducing vitamin D serum (p-value < 0.05). CONCLUSION: Higher micronutrient intake and higher vitamin D serum levels seem to reduce Chronic Oral Diseases Burden. Healthy diet policies may jointly tackle caries, periodontitis, obesity, and other non-communicable diseases.

Salivary cytokine expression after non-surgical periodontal therapy in smokers: 12-month follow-up.

BACKGROUND: Diverse smoking trajectories may influence cytokine expression after non-surgical periodontal therapy and supportive periodontal care. Thus, we aimed to describe cytokine profiles in periodontal healing after periodontal therapy in smokers. METHODS: A smoking cessation program and non-surgical periodontal therapy were offered to 80 smokers with periodontitis. Smoking trajectories (quitters/light, moderate, heavy) were observed. The association of salivary interleukin (IL)-1ß, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13, interferon-gamma (IFN-γ), and tumor necrosis factor-alpha (TNF-α) with smoking trajectories and periodontal outcomes was determined using mixed-effects linear regression. RESULTS: Among quitters/light smokers, IL-1ß was associated with an increase in mean periodontal pocket depth (PPD) and mean clinical attachment level (CAL). IL-6 was associated with a decrease in mean PPD and CAL in heavy smokers, whereas IL-8 was associated with a decrease in PPD among moderate smokers. TNFα was associated with a reduction in mean PPD and CAL among quitters/light smokers, while among moderate smokers, TNFα was associated with an increase in mean PPD and CAL. IL-12 and IL-13 were associated with a decrease in mean PPD in moderate smokers. CONCLUSION: Our findings suggest that distinctive smoking exposures induce differential cytokine expression, which, in turn, seems to influence periodontal repair.

Precursors of insulin resistance underlying periodontitis in adolescents aged 17-18 years.

OBJECTIVE: To investigate the association between insulin resistance markers and periodontitis in adolescents, analyzing confounder variables and the adiposity as a mediator. METHODS: This is population-based study is representative of adolescents aged 17-18 years from public schools in São Luís, Brazil (n = 405). Insulin resistance was assessed using the Model of Assessment of the Homeostasis of the Insulin Resistance Index (HOMA-IR) and its percussor triglycerides/HDL-cholesterol ratio (TG/HDL-c). The outcome was Initial Periodontitis, a latent variable estimated by the common variance shared among bleeding on probing, probing depth ≥ 4 mm, and clinical attachment loss ≥ 4 mm. The association between insulin resistance and Initial Periodontitis was modeled via pathways triggered by socioeconomic status, smoking, alcohol, and Adiposity, using structural equation modeling. RESULTS: Higher TG/HDL-c was directly associated with higher Initial Periodontitis (standardized coefficient [SC] = 0.130, p < 0.001). HOMA-IR was not associated with periodontal outcome (SC = 0.023, p = 0.075), but it was with Adiposity (SC = 0.495, p < 0.001). Higher TG/HDL-c was associated with Adiposity (SC = 0.202, p < 0.001). CONCLUSION: The insulin resistance markers were associated with early signs of periodontal breakdown among adolescents, suggesting a possible relationship between diabetes and periodontitis commences early in life.

Cytokine profiles and the dynamic of gingivitis development in humans.

AIM: To investigate the relationship between cytokine profiles and "fast" and "slow" patterns of gingival inflammation development. MATERIALS AND METHODS: Forty-two adults participated in an experimental gingivitis study, comprising a 2-week hygiene phase (clinical examination and professional cleaning); a 3-week induction phase (absence of oral hygiene); and a 2-week resolution phase (re-establishment of oral hygiene). Plaque and gingival inflammation scores were assessed. Interferon-gamma (IFN-γ), interleukin (IL)-1ß, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13, and tumour necrosis factor-alpha (TNF-α) from gingival crevicular fluid were collected and measured by multiplex ELISA. Group-based-trajectory-modelling (GBTM) was used to model cytokine profiles over the induction phase. The effect of gingival inflammation on cytokine levels over time was estimated with mixed-effects modelling. RESULTS: GBTM analysis revealed two cytokine profiles, "non-organized response" (IL-4, IL-6, IL-8, IL-12, and IL-13) and "organized response" (IL-2, IL-10, and TNF-α). Among the "slow" responders, neither cytokine profile was associated with gingivitis. In contrast, a "fast" response was associated with a higher "non-organized response" factor (coef. 0.14) and a lower "organized response" factor (coef. -0.03). CONCLUSION: A "fast" gingivitis development was associated with a higher "non-organized response" and a lower "organized response", which may elucidate the role of individual variability in gingivitis susceptibility.

Dietary vitamin D and calcium and periodontitis: A population-based study.

Aim: This study aimed to explore the relationship between dietary vitamin D and calcium intake and periodontitis among adults and whether it differs from males to females. Methods: Cross-sectional analysis of a population-based cohort study with adults aged 20 to 60 from Southern Brazil. Intake of vitamin D and calcium were gathered in 2012 using two 24h-dietary recalls. Clinical examination assessed the clinical attachment level and bleeding on probing. Confounders included sex, age, family income, smoking, and obesity. The controlled direct effect of vitamin D and calcium on periodontitis was examined using marginal structural modeling. Analyses were also stratified by sex. Results: Of the 1,066 investigated adults (mean age 35 ± 11.7 years; 49% females), 12.3% (95%CI 10.2;14.7) had periodontitis. Calcium intake had a direct protective effect on periodontitis (risk ratio (RR) 0.61; 95%CI 0.45;0.83), whereas no association between vitamin D and periodontitis was observed (RR 1.13; 95%CI 0.82;1.56). Stratified analyses revealed a null association between both vitamin D and calcium intake and periodontitis among men, but a protective association between calcium and intake and periodontitis among women (RR 0.56; 95%CI 0.38;0.79), while vitamin D remained without any association (RR 1.07; 95%CI 0.72;1.61). Conclusion: Our findings suggest a protective association between dietary calcium intake and periodontitis among women.

Influence of Bacterial Profiles in Cytokine and Clinical Features of Endodontic Disease.

INTRODUCTION: We verified the association between selected bacterial profiles and levels of cytokines, chemokines, and the expression of signs and symptoms of primary endodontic infection with apical periodontitis. METHODS: Samples were collected from 21 root canals, and macrophages were stimulated for 24 hours. Tumor necrosis factor alpha (TNF-α), interleukin (IL)-6, IL-10, IL-12p70, interferon gamma, and chemokine (C-C motif) ligand 2 (CCL2) were measured using cytometric bead array. We investigated the overlapping networks between cytokines and chemokines with regression analysis. Checkerboard DNA-DNA hybridization was used to assess 40 target bacteria species. Using factor analysis, bacterial species aggregated in 2 factors. The association of bacteria species-based factors on cytokine and chemokine levels and clinical features was estimated with regression analysis. RESULTS: A negative relationship between IL-10 (anti-inflammatory cytokine) and CCL2, TNF-α, and IFN-γ (proinflammatory cytokines) (all P < .05) was observed. CCL2 was positively correlated with TNF-α (P < .01). Thirty-eight bacteria species were detected in primary endodontic infection with apical periodontitis. The first bacteria species-based factor was associated with the size of the radiolucent area (coefficient = 15.42) and tenderness to percussion/pain on palpation (coefficient = 20.79). The second factor was associated with CCL2 levels (coefficient = 1.28). CONCLUSIONS: Different bacterial profiles can be differentially related to the expression of inflammatory proteins and the experience of clinical features.

Collider bias in the association of periodontitis and carotid intima-media thickness.

OBJECTIVES: This cross-sectional study tested the presence of collider bias in the relationship between periodontitis and the carotid intima-media thickness (cIMT). METHODS: Data from 480 members of the 1982 Pelotas Birth Cohort, Brazil, were used. Periodontitis at the age of 24 years was determined as the main exposure. cIMT at the age of 30 years was set as the outcome. High-sensitivity C-reactive protein (hsCRP) was considered the mediator (collider). Confounding variables included sex, income, BMI and smoking. The association between cIMT and periodontitis was tested in conventional logistic regression stratified on hsCRP levels, marginal structural modelling and sensitivity analysis for collider stratification bias. RESULTS: Conventional adjusted logistic regression analysis showed a positive association between periodontitis and cIMT (OR 1.5; 95% CI 1.1; 2.3). Stratified analysis according to the hsCRP levels revealed that the magnitude of the association was even higher among participants with hsCRP ≥ 3 mg/L (OR 2.2, 95% CI 1.1; 4.2) with 36% collider bias probability. No association between periodontitis and cIMT was found among participants with hsCRP < 3 mg/L (OR 1.3; 95% CI 0.8; 2.1). The association was not detected using marginal structural modelling (OR 1.3, 95% CI 0.8; 2.0). CONCLUSIONS: The association between periodontitis and surrogate markers of cardiovascular disease might be induced by collider bias stratification using conventional regression analysis.

Periodontitis and self-reported halitosis among young adults from the 1982 Pelotas Birth Cohort.

OBJECTIVE: To investigate the association between periodontitis and self-reported halitosis among young adults, and whether there is an interaction between the effects of smoking and periodontitis on halitosis. METHODS: Data from the 1982 Pelotas birth cohort, Brazil, were used. The controlled direct effect of periodontitis on self-reported halitosis, not mediated by tongue coating, was estimated using marginal structural modeling. In addition, an interaction between the effects of smoking and periodontitis on halitosis was also tested. Confounders comprised sociodemographic information, obesity, diabetes, and oral hygiene habits. RESULTS: The controlled direct effect of periodontitis on halitosis not mediated by tongue coating showed that individuals with periodontitis had 90% higher risk of self-reporting halitosis (RR 1.90) compared to healthy individuals. Individuals with mild periodontitis had twice the risk of reporting halitosis than periodontally healthy individuals (RR 2.31). We also found an interaction between the effects of smoking and periodontitis on halitosis, as noted among smokers with mild (RR 2.91) and moderate-to-severe periodontitis (RR 5.84). CONCLUSION: There is a controlled direct effect of periodontitis on halitosis not mediated by tongue coating. Additionally, an interaction between the effects of smoking and periodontitis on halitosis was also detected.

Impact of Smoking Cessation on Periodontitis: A Systematic Review and Meta-analysis of Prospective Longitudinal Observational and Interventional Studies.

AIMS: This systematic review aimed to estimate the effect of tobacco smoking cessation on the risk for periodontitis compared to the risk among never-smokers and to evaluate the effect of tobacco smoking cessation on the clinical outcomes of nonsurgical periodontal treatment. METHODS: Electronic searches were performed in PubMed, Scopus, and Embase. Search strategy included MeSH and free terms: periodontitis, periodontal diseases, smoking, tobacco use, tobacco, tobacco products, cigarette, pipe, and cigar. Only original prospective longitudinal observational and interventional studies that investigated the association between smoking cessation and periodontitis onset or progression were included. Meta-analyses were conducted to summarize the evidence. RESULTS: A total of 2743 articles were identified in electronic searches; out of which only six were included in the meta-analysis. Pooled estimates showed that the risk of periodontitis incidence or progression among those who quit smoking was not significantly different from the risk for never-smokers (risk ratio [RR] = 0.97; 95% confidence interval [CI] = 0.87% to 1.08%). Smokers had approximately 80% higher risk of periodontitis than quitters (RR = 1.79; 95% CI = 1.36% to 2.35%) and never-smokers (RR = 1.82; 95% CI = 1.43% to 2.31%). Periodontal therapy resulted in up to 0.2 mm (95% CI = -0.32% to -0.08%) higher gain in attachment level and extra 0.32 mm (95% CI = 0.07% to 0.52%) reduction in pocket depth among quitters over nonquitters after short follow-up (12-24 months). CONCLUSIONS: Few studies on the topic were identified. Smoking cessation reduced the risk for periodontitis onset and progression, and improved the outcomes of nonsurgical periodontal therapy. IMPLICATIONS: This review provides the first quantitative evidence of the impact of smoking cessation on the risk for periodontitis onset and progression. The findings have demonstrated that the risk for periodontitis becomes comparable to that of never-smokers and that nonsurgical periodontal treatment outcomes improve after smoking cessation. Dental professionals ought to consider smoking cessation interventions as a relevant component of the periodontal therapy.

Metabolic syndrome and periodontitis: A structural equation modeling approach.

BACKGROUND: This study aimed to investigate the association between metabolic syndrome (MetS) and periodontitis among young adults, and also to compare results using observed and latent variables for MetS and periodontitis. METHODS: Data from the 1982 Pelotas Birth Cohort, Brazil, were used. Metabolic syndrome at the age of 23 years was measured using clinical and biochemical analysis and set as the main exposure. Periodontitis at the age of 31 years was clinically measured and set as the outcome. Confounding variables included sex and maternal education, assessed at birth, family income at 23 years, and smoking status at the age of 23 and 30 years. Factor analyses (exploratory and confirmatory) were performed to define latent variables for MetS and periodontitis. In addition, both conditions were also defined as categorical observed variables. The association between MetS and periodontitis was tested in structural equation models. RESULTS: Two latent periodontal variables were identified: "initial" and "advanced" periodontitis, while one latent variable was identified for MetS. Metabolic syndrome is positively associated with "advanced" (coefficient 0.11; P value < 0.01), but not with "initial" (coefficient -0.01; P value = 0.79) periodontitis. When MetS and periodontitis were set as observed variables in the structural equation models, no association was found irrespective of the criteria used for periodontitis classification. CONCLUSIONS: There was a positive association between metabolic syndrome and "advanced" periodontitis, when the multiple dimensions of both diseases were accounted in latent variables. Nevertheless, when MetS and periodontitis were treated as observed variables, no association was detected irrespective of the criteria used for periodontitis classification.

Pattern recognition receptor polymorphisms in early periodontitis.

BACKGROUND: Even though bacteria trigger inflammation, most of the tissue destruction in periodontitis is due to the host inflammatory response. In addition to immunological events that drive development of early periodontitis, numerous environmental factors like genetics and smoking play a role. We investigated whether the carriage of selected single nucleotide polymorphisms (SNP) of toll-like receptors (TLR), NOD-like receptors (NLR) and RIG-I-like receptors (RLR) was associated with the diagnosis of early periodontitis in a case-control study. METHODS: Adolescents with positive (n = 87) and negative (n = 73) diagnosis for periodontitis had blood samples taken. All participants were genotyped for 42 SNP in the genes encoding TLR1-10, NOD1-2, DDX58, and IFIH1 using multiplex assays. Associations between SNP and periodontitis diagnosis were tested. RESULTS: TLR1-rs5743611 showed protective effect for periodontitis (CC versus GG and GC, P = 0.01, odds ratio [OR] 0.10, 95% confidence interval [CI] 0.01-0.78). Carriage of the TLR4-rs7873784 was associated with higher odds for periodontitis (GG versus CC and GC, P = 0.05, OR 2.30, 95% CI 1.00-5.63; GG versus GC, P = 0.05, OR 2.46, 95% CI 1.01-5.99). In male participants, reduced susceptibility to periodontitis was observed in carriers of TLR7-rs3853839 (CC versus GG and CG, P = 0.02, OR 0.30, 95% CI 0.11-0.85) and TLR8-rs3764879 (CC versus GG and CG, P = 0.02, OR 0.31, 95% CI 0.12-0.82). Associations were maintained after adjustments for sex, smoking habits, and mother´s education. CONCLUSION: This study demonstrated an association between TLR1-rs5743611, TLR4-rs7873784, TLR7-rs3853839, and TLR8-rs3764879 and susceptibility to periodontitis in adolescents.

Effect of Smoking on Periodontitis: A Systematic Review and Meta-regression.

CONTEXT: The study systematically reviewed articles on the association between tobacco smoking and periodontitis, as it has been hypothesized that smoking affects the course of periodontitis through impairment of immunological and vascular mechanisms. EVIDENCE ACQUISITION: Searches of articles indexed in PubMed, Scopus, and Embase were performed up to and including May 2017. Search strategy included MeSH and free terms: periodontitis, periodontal diseases, smoking, tobacco use, tobacco, tobacco products, cigarette, pipe, and cigar. Only original prospective longitudinal studies that investigated the association between smoking and periodontitis incidence or progression were included. Results were shown as combined risk ratio. Meta-regression and subgroup analyses were used to explore potential sources of heterogeneity. Analyses were conducted in August 2017. EVIDENCE SYNTHESIS: Twenty-eight studies were included in the review; of these, only 14 presented data that could be included in the meta-analysis. Pooled adjusted risk ratios estimate that smoking increases the risk of periodontitis by 85% (risk ratio=1.85, 95% CI=1.5, 2.2). Meta-regression demonstrated that age explained 54.2% of the variability between studies, time of follow-up explained 13.5%, loss to follow-up 10.7%, criteria used to assess the periodontal status explained 2.1%, and severity of periodontitis explained 16.9%. CONCLUSIONS: Smoking has a detrimental effect on the incidence and progression of periodontitis. Tobacco smoking, therefore, is important information that should be assessed along with other risk factors for periodontitis.

Comparison of Fusobacterium nucleatum and Porphyromonas gingivalis Lipopolysaccharides Clinically Isolated from Root Canal Infection in the Induction of Pro-Inflammatory Cytokines Secretion

Abstract The aim of this study was to compare the biological activity of lipopolysaccharides (LPS) purified from Fusobacterium nucleatum and Porphyromonas gingivalis strains, both isolated from primary endodontic infection (PEI) in the levels of IL-1β and TNF-α released by macrophage cells. Moreover, LPS was purified from F. nucleatum and P. gingivalis American Type Collection (ATCC) and its biological activity was compared to respectively clinical isolates strains. F. nucleatum and P. gingivalis strains clinically isolated from PEI had their identification confirmed by sequencing the 16S rRNA gene. LPS from F. nucleatum and P. gingivalis and their respective ATCC strains were extracted by using Tri-reagent method. Macrophages (Raw 264.7) were stimulated with LPS at 100 ng/mL for 4, 8 and 12 h. Secretion of IL-1 β and TNF-α was also determined. Paired t-test, repeated measures ANOVA and one-way ANOVA were employed. All LPS induced significant production of IL-1β and TNF-α, with the former being secreted at higher levels than the latter in all time-points. F. nucleatum induced a higher expression of both cytokines compared to P. gingivalis (p<0.05). No differences were observed between clinical and ATCC strains, as both presented the same potential to induce pro-inflammatory response. It was concluded that F. nucleatum and P. gingivalis LPS presented different patterns of activation against macrophages as seen by the IL-1β and TNF-α production, which may contribute to the immunopathogenesis of apical periodontitis. Moreover, clinical and ATCC strains grown under the same in vitro environment conditions presented similar biological activity.

Resumo O objetivo deste estudo foi comparar a atividade biológica de lipopolissacarídeos (LPS) purificados a partir de linhagens de Fusobacterium nucleatum e Porphyromonas gingivalis, ambas isoladas de infecções endodônticas primárias (IEP) nos níveis de IL-1β e TNF-α produzidos por macrófagos. Adicionalmente, LPS foi purificado de F. nucleatum e P. gingivalis "American Type Collection" (ATCC) e sua atividade comparada às respectivas linhagens clinicamente isoladas. Linhagens de F. nucleatum e P. gingivalis isoladas clinicamente de IEP tiveram sua identificação confirmada por sequenciamento do gene 16S rRNA. LPS de F. nucleatum e P. gingivalis e das respectivas linhagens foram extraídos com o uso do método "Tri-reagent". Macrófagos (Raw 264.7) foram estimulados com LPS a 100 ng/mL por 4, 8 e 12 h. A secreção de IL-1β e de TNF-α foi determinada. Foram usados os testes t-pareado, ANOVA de medidas repetidas e ANOVA de um fator. Todos os LPS induziram a produção significante de IL-1β e TNF-α, sendo o primeiro secretado em mais altas concentrações que o último em todos os tempos avaliados. F. nucleatum induziu uma maior expressão de ambas as citocinas comparativamente ao P. gingivalis (p<0,05). Não foram observadas diferenças entre as linhagens clínica e ATCC, uma vez que ambas apresentaram o mesmo potencial de indução da resposta pró-inflamatória. Conclui-se que F. nucleatum e P. gingivalis possuem diferentes padrões de ativação dos macrófagos, como visto pela produção de IL-1β e TNF-α, o que pode contribuir para a imunopatogênese da periodontite apical. Ainda, linhagens clínica e ATCC mantidas no mesmo ambiente in vitro apresentaram ativação biológica semelhante.

Proinflammatory Activity of Primarily Infected Endodontic Content against Macrophages after Different Phases of the Root Canal Therapy.

INTRODUCTION: This study investigated the presence of target bacterial species and the levels of endotoxins in teeth with apical periodontitis. Levels of inflammatory mediators (interleukin [IL]-1ß and tumor necrosis factor [TNF]-α) were determined after macrophage stimulation with endodontic content after different phases of endodontic therapy using different irrigants. METHODS: Thirty primarily infected root canals were randomly assigned into 3 groups according to the irrigant used for root canal preparation (n = 10 per group): GI: 2.5% sodium hypochlorite, GII: 2% chlorhexidine gel, and GIII (control group): saline solution. Root canal samples were taken by using paper points before (s1) and after root canal instrumentation (s2), subsequently to 17% EDTA (s3), after 30 days of intracanal medication (Ca[OH]2 + saline solution) (s4), and before root canal obturation (s5). Polymerase chain reaction (16S recombinant DNA) and limulus amebocyte lysate assay were used for bacterial and endotoxin detection, respectively. Macrophages were stimulated with the root canal contents for IL-1ß/TNF-α measurement using enzyme-linked immunosorbent assay. RESULTS: Porphyromonas gingivalis (17/30), Porphyromonas endodontalis (15/30), and Prevotella nigrescens (11/30) were the most prevalent bacterial species. At s1, endotoxins were detected in 100% of the root canals (median = 32.43 EU/mL). In parallel, substantial amounts of IL-1ß and TNF-α were produced by endodontic content-stimulated macrophages. At s2, a significant reduction in endotoxin levels was observed in all groups, with GI presenting the greatest reduction (P < .05). After a root canal rinse with EDTA (s3), intracanal medication (s4), and before root canal obturation (s5), endotoxin levels reduced without differences between groups (P < .05). IL-1ß and TNF-α release decreased proportionally to the levels of residual endotoxin (P < .05). CONCLUSIONS: Regardless of the use of sodium hypochlorite or CHX, the greatest endotoxin reduction occurs after chemomechanical preparation. Increasing steps of root canal therapy associated with intracanal medication enhances endotoxin reduction, leading to a progressively lower activation of proinflammatory cells such as macrophages.

Can peri-implant crevicular fluid assist in the diagnosis of peri-implantitis? A systematic review and meta-analysis.

BACKGROUND: A broader understanding of the immune inflammatory profile of peri-implant diseases could be helpful in the development of host-targeted preventive and therapeutic strategies. The aim of this study is to answer two clinical questions: 1) whether patients with peri-implantitis (PP) present higher prevalence of any specific inflammatory cytokine in peri-implant crevicular fluid (PICF) compared with healthy patients; and 2) whether local inflammation measured in PICF can be used as a predictor for incipient PP. METHODS: A systematic review of the literature on the most common cytokines released in PICF in healthy and PP-affected sites was conducted from 1996 up to and including October 2013 using predefined search strategies. Cross-sectional and prospective longitudinal studies were considered. Meta-analyses were done separately for healthy, mucositis (MU), and PP outcomes. RESULTS: Interleukin (IL)-1ß was the most studied cytokine (n = 12), followed by tumor necrosis factor (TNF)-α (n = 10). Other cytokines were also linked to PP, such as IL-4, IL-6, IL-8, IL-10, IL-12, and IL-17. Statistical differences were revealed when IL-1ß release was compared between healthy implant sites and PP (P = 0.001) or MU sites (P = 0.002), respectively; when PP and MU were compared, no statistical differences could be detected (P = 0.80). For TNF-α release, significant differences were found between healthy and PP implants (P = 0.02). CONCLUSIONS: PICF containing inflammatory mediators, such as IL-1ß and TNF-α, can be used as additional criteria for a more robust diagnosis of peri-implant infection. Additionally, once the inflammatory process is installed, no differences were found between peri-implant MU and PP.

Clinical influence of different intracanal medications on Th1-type and Th2-type cytokine responses in apical periodontitis.

INTRODUCTION: This clinical study assessed the influence of different intracanal medications on Th1-type and Th2-type cytokine responses in apical periodontitis and monitored the levels of bacteria from primarily infection during endodontic procedures. METHODS: Thirty primarily infected teeth were randomly divided into 3 groups according to the medication selected: chlorhexidine (CHX), 2% CHX gel; Ca(OH)2/SSL, Ca(OH)2 + SSL; and Ca(OH)2/CHX, Ca(OH)2 + 2% CHX gel (all, n = 10). Bacterial sample was collected from root canals, and the interstitial fluid was sampled from lesions. Culture techniques were used to determine bacterial counts (colony-forming units/mL). Th1 (tumor necrosis factor-α, interferon-γ, and interleukin [IL]-2) and Th2 cytokines (IL-4, IL-5, and IL-13) were measured by enzyme-linked immunosorbent assay. RESULTS: All intracanal medication protocols were effective in reducing the bacterial load from root canals (all P < .05) and lowering the levels of Th1-type cytokines in apical lesions (all P < .05), with no differences between them (P > .05). Both Ca(OH)2 treatment protocols significantly increased the levels of Th2-type cytokines (P < .05), with no differences between them (P > .05). Thus, chlorhexidine medication showed the lowest effectiveness in increasing the levels of Th2-type cytokine. After treatment, regardless of the type of medication, the linear regression analysis indicated the down-regulation of Th2-type cytokines by Th1-type cytokines. CONCLUSIONS: All intracanal medication protocols were effective in reducing bacterial load and lowering the levels of Th1-type cytokines. Thus, the use of Ca(OH)2 medications contributed to the increase in the Th2-type cytokine response in apical periodontitis.

Macrophage cell activation with acute apical abscess contents determined by interleukin-1 Beta and tumor necrosis factor alpha production.

INTRODUCTION: This clinical study has investigated the antigenic activity of bacterial contents from exudates of acute apical abscesses (AAAs) and their paired root canal contents regarding the stimulation capacity by levels of interleukin (IL)-1 beta and tumor necrosis factor alpha (TNF-α) throughout the root canal treatment against macrophage cells. METHODS: Paired samples of infected root canals and exudates of AAAs were collected from 10 subjects. Endodontic contents were sampled before (root canal sample [RCS] 1) and after chemomechanical preparation (RCS2) and after 30 days of intracanal medication with calcium hydroxide + chlorhexidine gel (Ca[OH]2 + CHX gel) (RCS3). Polymerase chain reaction (16S rDNA) was used for detection of the target bacteria, whereas limulus amebocyte lysate was used to measure endotoxin levels. Raw 264.7 macrophages were stimulated with AAA exudates from endodontic contents sampled in different moments of root canal treatment. Enzyme-linked immunosorbent assays were used to measure the levels of TNF-α and IL-1 beta. RESULTS: Parvimonas micra, Porphyromonas endodontalis, Dialister pneumosintes, and Prevotella nigrescens were the most frequently detected species. Higher levels of endotoxins were found in samples from periapical exudates at RCS1 (P < .005). In fact, samples collected from periapical exudates showed a higher stimulation capacity at RCS1 (P < .05). A positive correlation was found between endotoxins from exudates with IL-1 beta (r = 0.97) and TNF-α (r = 0.88) production (P < .01). The significant reduction of endotoxins and bacterial species achieved by chemomechanical procedures (RCS2) resulted in a lower capacity of root canal contents to stimulate the cells compared with that at RCS1 (P < .05). The use of Ca(OH)2 + CHX gel as an intracanal medication (RCS3) improved the removal of endotoxins and bacteria from infected root canals (P < .05) whose contents induced a lower stimulation capacity against macrophages cells at RCS1, RCS2, and RCS3 (P < .05). CONCLUSIONS: AAA exudates showed higher levels of endotoxins and showed a greater capacity of macrophage stimulation than the paired root canal samples. Moreover, the use of intracanal medication improved the removal of bacteria and endotoxins from infected root canals, which may have resulted in the reduction of the inflammatory potential of the root canal content.