Activation of the neutrophil respiratory burst by plasma from periodontitis patients is mediated by pro-inflammatory cytokines

Aim: To determine the effect of periodontitis patients' plasma on the neutrophil oxidative burst and the role of albumin, immunoglobulins (Igs) and cytokines. Materials and Methods: Plasma was collected from chronic periodontitis patients (n=11) and periodontally healthy controls (n=11) and used with/without depletion of albumin and Ig or antibody neutralization of IL-8, GM-CSF or IFN-a to prime/stimulate peripheral blood neutrophils, isolated from healthy volunteers. The respiratory burst was measured by lucigenin-dependent chemiluminescence. Plasma cytokine levels were determined by ELISA. Results: Plasmas from patients were significantly more effective in both directly stimulating neutrophil superoxide production and priming for subsequent formyl-met-leu-phe (fMLP)-stimulated superoxide production than plasmas from healthy controls (p<0.05). This difference was maintained after depletion of albumin and Ig. Plasma from patients contained higher mean levels of IL-8, GM-CSF and IFN-a. Individual neutralizing antibodies against IL-8, GM-CSF or IFN-a inhibited the direct stimulatory effect of patients' plasma, whereas the ability to prime for fMLP-stimulated superoxide production was only inhibited by neutralization of IFN-a. The stimulating and priming effects of control plasma were unaffected by antibody neutralization. Conclusions: This study demonstrates that plasma cytokines may have a role in inducing the hyperactive (IL-8, GM-CSF, IFN-a) and hyper-reactive (IFN-a) neutrophil phenotype seen in periodontitis patients.

Oxidative and inflammatory status in Type 2 diabetes patients with periodontitis

Aim: To determine the impact of periodontitis on oxidative/inflammatory status and diabetes control in Type 2 diabetes. Materials and Methods: A comparative study of 20 Type 2 diabetes patients with periodontitis [body mass index (BMI) 31+5], 20-age/gender-matched, non-periodontitis Type 2 diabetes controls (BMI 29+6) and 20 non-diabetes periodontitis controls (BMI 25+4) had periodontal examinations and fasting blood samples collected. Oxidative stress was determined by plasma small molecule antioxidant capacity (pSMAC) and protein carbonyl levels; inflammatory status by total/differential leucocytes, fibrinogen and high sensitivity C-reactive protein (hsCRP); diabetes status by fasting glucose, HbA1c, lipid profile, insulin resistance and secretion. Statistical analysis was performed using SPSS. Results: pSMAC was lower (p=0.03) and protein carbonyls higher (p=0.007) in Type 2 diabetes patients with periodontitis compared with those without periodontitis. Periodontitis was associated with significantly higher HbA1c (p=0.002) and fasting glucose levels (p=0.04) and with lower ß-cell function (HOMA-ß; p=0.01) in diabetes patients. Periodontitis had little effect on inflammatory markers or lipid profiles, but Type 2 diabetes patients with periodontitis had higher levels of hsCRP than those without diabetes (p=0.004) and the lowest levels of HDL-cholesterol of all groups. Conclusion: Periodontitis is associated with increased oxidative stress and compromised glycaemic control in Type 2 diabetes patients.

Sulforaphane restores cellular glutathione levels and reduces chronic periodontitis neutrophil hyperactivity in vitro

The production of high levels of reactive oxygen species by neutrophils is associated with the local and systemic destructive phenotype found in the chronic inflammatory disease periodontitis. In the present study, we investigated the ability of sulforaphane (SFN) to restore cellular glutathione levels and reduce the hyperactivity of circulating neutrophils associated with chronic periodontitis. Using differentiated HL60 cells as a neutrophil model, here we show that generation of extracellular O2 . - by the nicotinamide adenine dinucleotide (NADPH) oxidase complex is increased by intracellular glutathione depletion. This may be attributed to the upregulation of thiol regulated acid sphingomyelinase driven lipid raft formation. Intracellular glutathione was also lower in primary neutrophils from periodontitis patients and, consistent with our previous findings, patients neutrophils were hyper-reactive to stimuli. The activity of nuclear factor erythroid-2-related factor 2 (Nrf2), a master regulator of the antioxidant response, is impaired in circulating neutrophils from chronic periodontitis patients. Although patients' neutrophils exhibit a low reduced glutathione (GSH)/oxidised glutathione (GSSG) ratio and a higher total Nrf2 level, the DNA-binding activity of nuclear Nrf2 remained unchanged relative to healthy controls and had reduced expression of glutamate cysteine ligase catalytic (GCLC), and modifier (GCLM) subunit mRNAs, compared to periodontally healthy subjects neutrophils. Pre-treatment with SFN increased expression of GCLC and GCM, improved intracellular GSH/GSSG ratios and reduced agonist-activated extracellular O2 . - production in both dHL60 and primary neutrophils from patients with periodontitis and controls. These findings suggest that a deficiency in Nrf2-dependent pathways may underpin susceptibility to hyper-reactivity in circulating primary neutrophils during chronic periodontitis. © 2013 Dias et al.

High permeability of the anionic form restricts accumulation of indomethacin by cultured gastric surface epithelial cells exposed to low apical pH

The 'ion-trapping' hypothesis suggests that the intracellular concentration of acidic non-steroidal anti-inflammatory drugs in gastric epithelial cells could be much higher than in the gastric lumen, and that such accumulation could contribute to their gastrotoxicity. Our aim was to examine the effect of the pH of the apical medium on the apical to basal transfer of the acidic drug indomethacin (pK a 4.5) across a gastric mucous epithelial cell monolayer, and to determine whether indomethacin accumulated in cells exposed to a low apical pH. Guinea-pig gastric mucous epithelial cells were grown on porous membrane culture inserts (Transwells®) for 72 h. Transfer and accumulation of [ 14C] indomethacin were assessed by scintillation counting. Transfer of [ 3H]mannitol and measurement of trans-epithelial electrical resistance were used to assess integrity of the monolayer. Distribution of [ 14C] urea was used to estimate the intracellular volume of the monolayer. The monolayer was not disrupted by exposure of the apical face to media of pH ≥ 3, or by indomethacin. Transfer of indomethacin (12 μM) to the basal medium increased with decreasing apical medium pH. The apparent permeability of the undissociated acid was estimated to be five times that of the anion. The intracellular concentration of indomethacin was respectively 5.3, 4.1 and 4.3 times that in the apical medium at pH 5.5, 4.5 and 3.0. In conclusion, this study represents the first direct demonstration that indomethacin accumulates in gastric epithelial cells exposed to low apical pH. However, accumulation of indomethacin was moderate and the predictions of the ion-trapping hypothesis were not met, probably due to the substantial permeability of anionic indomethacin across membranes. © 2006 Elsevier B.V. All rights reserved.

Serum α-tocopherol has a nonlinear inverse association with periodontitis among US adults

Background: Previous experimental models suggest that vitamin E may ameliorate periodontitis. However, epidemiologic studies show inconsistent evidence in supporting this plausible association. Objective: We aimed to investigate the association between serum α-tocopherol (αT) and γ-tocopherol (γT) and periodontitis in a large cross-sectional US population. Methods: This study included 4708 participants in the 1999–2001 NHANES. Serum tocopherols were measured by HPLC and values were adjusted by total cholesterol (TC). Periodontal status was assessed by mean clinical attachment loss (CAL) and probing pocket depth (PPD). Total periodontitis (TPD) was defined as the sum of mild, moderate, and severe periodontitis. All measurements were performed by NHANES. Results: Means ± SDs of serum αT:TC ratio from low to high quartiles were 4.0 ± 0.4, 4.8 ± 0.2, 5.7 ± 0.4, and 9.1 ± 2.7 μmol/mmol. In multivariate regression models, αT:TC quartiles were inversely associated with mean CAL (P-trend = 0.06), mean PPD (P-trend < 0.001), and TPD (P-trend < 0.001) overall. Adjusted mean differences (95% CIs) between the first and fourth quartile of αT:TC were 0.12 mm (0.03, 0.20; P-difference = 0.005) for mean CAL and 0.12 mm (0.06, 0.17; P < 0.001) for mean PPD, whereas corresponding OR for TPD was 1.65 (95% CI: 1.26, 2.16; P-difference = 0.001). In a dose-response analysis, a clear inverse association between αT:TC and mean CAL, mean PPD, and TPD was observed among participants with relatively low αT:TC. No differences were seen in participants with higher αT:TC ratios. Participants with γT:TC ratio in the interquartile range showed a significantly lower mean PPD than those in the highest quartile. Conclusions: A nonlinear inverse association was observed between serum αT and severity of periodontitis, which was restricted to adults with normal but relatively low αT status. These findings warrant further confirmation in longitudinal or intervention settings.

Serum α-tocopherol has a nonlinear inverse association with periodontitis among US adults

Previous experimental models suggest that vitamin E may ameliorate periodontitis. However, epidemiologic studies show inconsistent evidence in supporting this plausible association. We aimed to investigate the association between serum α-tocopherol (αT) and γ-tocopherol (γT) and periodontitis in a large cross-sectional US population. This study included 4708 participants in the 1999–2001 NHANES. Serum tocopherols were measured by HPLC and values were adjusted by total cholesterol (TC). Periodontal status was assessed by mean clinical attachment loss (CAL) and probing pocket depth (PPD). Total periodontitis (TPD) was defined as the sum of mild, moderate, and severe periodontitis. All measurements were performed by NHANES. Means ± SDs of serum αT:TC ratio from low to high quartiles were 4.0 ± 0.4, 4.8 ± 0.2, 5.7 ± 0.4, and 9.1 ± 2.7 μmol/mmol. In multivariate regression models, αT:TC quartiles were inversely associated with mean CAL (P-trend = 0.06), mean PPD (P-trend < 0.001), and TPD (P-trend < 0.001) overall. Adjusted mean differences (95% CIs) between the first and fourth quartile of αT:TC were 0.12 mm (0.03, 0.20; P-difference = 0.005) for mean CAL and 0.12 mm (0.06, 0.17; P < 0.001) for mean PPD, whereas corresponding OR for TPD was 1.65 (95% CI: 1.26, 2.16; P-difference = 0.001). In a dose-response analysis, a clear inverse association between αT:TC and mean CAL, mean PPD, and TPD was observed among participants with relatively low αT:TC. No differences were seen in participants with higher αT:TC ratios. Participants with γT:TC ratio in the interquartile range showed a significantly lower mean PPD than those in the highest quartile. A nonlinear inverse association was observed between serum αT and severity of periodontitis, which was restricted to adults with normal but relatively low αT status. These findings warrant further confirmation in longitudinal or intervention settings.

Apicectomías de dientes premolares y molares mandibulares en el conejo de raza neo zelandesa: estudio histológico usando distintos materiales de obturación apical

La apicectomía es una técnica de cirugía odontológica humana y veterinaria, que consiste en la amputación o resección de la porción más apical de la raíz dentaria y la retrobturación del ápice con algún material, con el objetivo de conservar el diente afectado de alguna patología apical. Se han usado gran variedad de materiales para la obturación, como amalgama, gutapercha, ionómero de vidrio, composite, MTA e hidróxido de calcio. En este estudio pretendemos comprobar la viabilidad de la técnica de la realización de la apicectomías en premolares y molares mandibulares, usando distintos materiales de retrobturación como son el iónomero de vidrio Ketac Cem μ® y el composite Tetric EvoFlow® en el conejo de experimentación de raza Neozelandesa. Realizamos las apicectomías de los dientes 407, 408 y 409 (3º premolar, 4º premolar y 1º molar inferior derecho), en una sola fase a diez animales. En el primer diente (407) solo se aplica un sellado del diente apicectomizado con hidróxido de calcio en polvo® y pasta Dycal® Dentin y no se coloca ningún material de obturación. En el segundo diente (408), se aplica un sellado con hidróxido de calcio en polvo® y pasta Dycal® Dentin y encima una retrobturación con ionómero de vidrio Ketac Cem μ® y en el tercer diente (409), se aplica un sellado con hidróxido de calcio en polvo® y pasta Dycal® Dentin y encima una retroturación con el composite Tetric EvoFlow®. A las 4 y 8 semanas se sacrificaron los animales y se realizó la toma de muestras. Estas fueron radiografiadas y posteriormente se realizaron cortes histológicos de los dientes apicectomizados y del tejido alrededor. Los cortes fueron teñidos con la tinción Levai-Laczkó y se realizó el estudio histológico cuantitativo y semicuantitativo, según la norma UNE-EN ISO 10993-6:2007, para medir la presencia de inflamación, fibrosis, infiltrados adiposos y cierre óseo del defecto...

Systemic inflammatory impact of periodontitis on acute coronary syndrome

Aim: A causative relationship between acute coronary syndrome (ACS) and periodontitis has yet to be defined. The aim of this study was to assess differences in levels of serum cytokines between individuals with or without ACS or periodontal comorbidity. Material and Methods: In a case–control study, individuals with ACS (78 individuals, 10.3% females) and matching healthy controls (78 individuals, 28.2% females) were included. Medical and dental examinations were performed to diagnose ACS and periodontitis. Serum levels of cytokines were assessed, using Luminex technology. Results: A diagnosis of periodontitis in the ACS and control group was diagnosed in 52.6% and 12.8% of the individuals, respectively. The unadjusted odds-ratio that individuals with ACS also had periodontitis was 7.5 (95% CI: 3.4, 16.8, p 

Análise comparativa "in vitro" da extrusão apical de detritos originados por diferentes técnicas de instrumentação do sistema de canais radiculares

Introdução: A extrusão apical detritos (EAD) consequência indesejável da instrumentação canalar pode ser associada a dor/edema, podendo atrasar a cicatrização periapical. O nosso trabalho teve como objectivo avaliar e quantificar a EAD em canais instrumentados por sistemas de instrumentação rotatória contínua e reciprocante. Materiais e Métodos: 80 dentes monocanalares sem tratamento endodôntico prévio foram aleatoriamente divididos em 4 grupos (n=20): One Shape® Protaper® NEXT, Hyflex® EDM e WaveOne® Gold. Um tubo de Eppendorf (TdE) foi pesado antecipadamente numa balança analítica de precisão e com um dente inserido foi montado num dispositivo modificado, similar ao método descrito por Myers & Montgomery. Os canais foram instrumentados e irrigados com água destilada. Os dentes instrumentados foram removidos dos TdE e estes preenchidos com água destilada até perfazer 1,5ml, incubados a 70ºC durante cinco dias sendo pesados novamente, calculando a diferença entre o peso inicial e final determinando o peso dos detritos. Os dados foram analisados estatisticamente utilizando o IBM SPSS Statistics 22, considerando α=0,05. Efetuaram-se testes Kruskal-Wallis e post-hoc com ajustamento do ρ-value pelo método Dunn-Bonferroni. Resultados: Houve EAD em todas as técnicas de instrumentação. A análise estatística mostrou haver diferenças significativas na EAD entre as técnicas utilizadas (α=0,002). Entre as técnicas WaveOne® Gold e One Shape® (α=0,003), WaveOne® Gold e Protaper® NEXT (α=0,023) e WaveOne® Gold e Hyflex® EDM (α=0,028). Conclusões: A técnica One Shape® apresentou menor EAD e a técnica WaveOne® Gold com movimento reciprocante constitui maior fator de risco tendo apresentado maior EAD. Os resultados deste estudo indicam que os profissionais devem estar cientes para a EAD que pode ocorrer com cada instrumento, o que poderá servir de base para a selecção de um instrumento particular. Implicações clínicas: A escolha do sistema de instrumentação canalar influencia a extrusão de detritos. Fontes de financiamento: Agradecimentos as empresas; Micro-Mega, França, COLTÉNE e Dentsply Maillefer, Suíça.

Prevalencia de la periodontitis y factores de riesgo en los trabajadores de la compañía GUAPAN, entre agosto del 2002 y marzo del 2003

Se estudia la prevalencia de la periodontitis con el íncice periodontal CPITN [Indice Comunitario de Necesidades de Tratamiento Periodontal] recomendado por la Organización Mundial de la Salud [OMS] y factores de riesgo en 243 trabajadores entre 25-76 años de edad, de la Compañía Industria Guapán S.A. [CIGSA], ubicada en la ciudad de Azogues, provincia del Cañar, Ecuador, Sur América. El trabajo fue aprobado por el Comité de Etica de la Facultad de Ciencias Médicas de la Universidad de Cuenca y el consentimiento se recibió en forma verbal. La prevalencia de la periodontitis fue de 25.5 por ciento; luego se analizó la relación entre la presentación o no de la periodontitis y la exposición o no al factor de riesgo; la relación entre el factor y la periodontitis, así como, el grado de esa asociación se determinó mediante el parámetro razón de la prevalencia [RP], con su respectivo IC al 95 por ciento, obteniendo los siguientes resultados: Mal hábito de higiene oral RP 4.71, IC 95 por ciento, 1.20-18.37; incremento de edad RP 1.83, IC 95 por ciento 1.09-3.10; bajo nivel de instrucción RP 1.89, IC 95 por ciento 1.07-3.34; prótesis parcial removible defectuosa [PPRD] RP 1.75, IC 95 por ciento 1.13-2.69; y, hábito de fumar RP 1.59, IC 95 por ciento 1.02-2.46; por tanto, al relacionar la periodontitis con los factores de riesgo estudiados, se encontró una asociación significativa con cada uno de ellos

Periodontitis as a risk factor for systemic disease: Are microparticles the missing link?

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Exogenous Administration of 15d-PGJ(2)-Loaded Nanocapsules Inhibits Bone Resorption in a Mouse Periodontitis Model

The 15-deoxy-(Delta 12,14)-PG J(2) (15d-PGJ(2)) has demonstrated excellent anti-inflammatory results in different experimental models. It can be used with a polymeric nanostructure system for modified drug release, which can change the therapeutic properties of the active principle, leading to increased stability and slower/prolonged release. The aim of the current study was to test a nano-technological formulation as a carrier for 15d-PGJ(2), and to investigate the immunomodulatory effects of this formulation in a mouse periodontitis model. Poly (D, L-lactide-coglycolide) nanocapsules (NC) were used to encapsulate 15d-PGJ(2). BALB/c mice were infected on days 0, 2, and 4 with Aggregatibacter actinomycetemcomitans and divided into groups (n = 5) that were treated daily during 15 d with 1, 3, or 10 mu g/kg 15d-PGJ(2)-NC. The animals were sacrificed, the submandibular lymph nodes were removed for FACS analysis, and the jaws were analyzed for bone resorption by morphometry. Immunoinflammatory markers in the gingival tissue were analyzed by reverse transcriptase-quantitative PCR, Western blotting, or ELISA. Infected animals treated with the 15d-PGJ(2)-NC presented lower bone resorption than infected animals without treatment (p < 0.05). Furthermore, infected animals treated with 10 mu g/kg 15d-PGJ(2)-NC had a reduction of CD4(+)CD25(+)FOXP3(+) cells and CD4/CD8 ratio in the submandibular lymph node (p < 0.05). Moreover, CD55 was upregulated, whereas RANKL was downregulated in the gingival tissue of the 10 mu g/kg treated group (p < 0.05). Several proinflammatory cytokines were decreased in the group treated with 10 mu g/kg 15d-PGJ(2)-NC, and high amounts of 15d-PGJ(2) were observed in the gingiva. In conclusion, the 15d-PGJ(2)-NC formulation presented immunomodulatory effects, decreasing bone resorption and inflammatory responses in a periodontitis mouse model. The Journal of Immunology, 2012, 189: 1043-1052.

Determinación del tamaño de la preparación apical: un dilema clínico en endodoncia

La determinación del tamaño de la preparación apical es un tema de constante debate, ya que a pesar de la evidencia científica disponible aún no se logra un consenso para saber cuál es el diámetro adecuado para obtener un tratamiento exitoso; en este sentido, el clínico debe ser crítico ante cada tratamiento, evaluando el diagnóstico y la anatomía del sistema de conductos radiculares. En la presente revisión, se realizó una búsqueda de la literatura disponible, con el fin de ofrecer al odontólogo un enfoque ante sus procedimientos clínicos, considerando diferentes parámetros, así como las distintas implicaciones clínicas que derivan de la determinación del tamaño de la preparación apical, lo cual constituye un aspecto importante dentro del proceso de la preparación biomecánica.

Reabsorción de hueso alveolar en pacientes diabéticos tipo 2 y pacientes no diabéticos con periodontitis crónica

Introducción: Existen diversos trastornos sistémicos que se manifiestan en la cavidad bucal, siendo el periodonto uno de los componentes que se ven más afectados. La Diabetes Mellitus es uno de los trastornos sistémicos que más se han estudiado en su relación con manifestaciones bucales. La microangiopatía, la alteración de la síntesis del colágeno y la disminuída acción fagocitaria hacen de las personas que padecen esta enfermedad sean más propensos a la Enfermedad Periodontal. Las enfermedades periodontales comúnmente abarcan númerosos y complejos signos y síntomas, motivo por el cual existen diversos métodos diagnósticos para evaluar la condición y magnitud de éstas. El indicador más importante de la magnitud con que se presenta la enfermedad periodontal está basado en la reabsorción de hueso alveolar, por ser este el tejido periodontal con evolución más lenta y por ser el que en última instancia se ve afectado. Tanto diabetes como enfermedad periodontal tienen una alta incidencia en la población general, ambas son multifactoriales y tienen que ver con alguna disfunción inmunoreguladora numerosos estudios indican mayor incidencia de enfermedad periodontal en personas con diabetes que en personas sanas. Objetivo: Determinar la influencia de la Diabetes Mellitus tipo 2 en el grado de reabsorción de hueso alveolar en pacientes con Periodontitis Crónica. Materiales y Métodos: En este estudio se incluyeron 17 pacientes diabéticos pertenecientes al Posgrado de Periodoncia de la facultad de Odontología de la Universidad Autónoma de Nuevo León y 29 pacientes no diabéticos que acudierón al Posgrado de Periodoncia de la Facultad de Odontología de la Universidad de Nuevo León, siguiendo los criterios de inclusión y exclusión. Parámetros clínicos fueron tomados en consideración para evaluar el estado periodontal tales como: profundidad de bolsa (PD), pérdida de inserción clínica (CAL), índice periodontal (PI) y evaluación radiográfica. De todos los pacientes que integraron ambos grupos. Se utilizó una rejilla milímetrada adosada a la placa radiográfica, con la cual se obtuvo una imagen radiopaca cuadriculada milímetrada, con el fin de facilitar una medición más exacta. Resultados: La investigación revela que, existe diferencia en el grado de enfermedad periodontal, representado por la reabsorción de hueso alveolar, entre el grupo de estudio (diabéticos tipo 2) = 4.81mm. promedio y el grupo control (no diabéticos) = 2.69 mm. promedio por otra parte, se encontró , diferencia estadísticamente significativa al comparar los promedios de reabsorción ósea alveolar total (p =<0.05). Conclusión: Dentro de las limitaciones del estudio, es posible asumir que diabetes mellitus no causa enfermedad periodontal, más bien crea condiciones para su proliferación.