Microbiological study of the subgingival biofilm in HIV+/HAART patients at a specialized dental service / Estudio microbiológico del biofilm subgingival de pacientes VIH+ bajo TARGA en un servicio dental especializado

ABSTRACT The aim of this study was to describe the microbiological profile of HIV patients under highly active antiretroviral treatment (HAART). This crosssectional study comprised 32 HIV patients with periodontal disease (PD) who had been under HAART for more than 6 months. Information about the patients' medical history was obtained from clinical records. Clinical dental examination was performed by a calibrated researcher using standard dental instruments to determine probing depth (PD), clinical attachment level (CAL), and bleeding on probing (BOP). A total 4,765 periodontal sites were evaluated, 125 of which were also studied microbiologically. Subgingival biofilm samples were obtained using sterile paper points; one set was used for microbiological culture studies and the other for endpoint PCR. Statistical analysis was performed using KruskalWallis and posthoc DunnBonferroni contrast tests. All participants were on HAART at the time of the study, and 90.6% had a viral load below 50 copies/mm3. Prevalence of periodontally active sites was low in the study population. Microbiological studies: Black pigmented anaerobic bacteria and fusiform CFU counts were significantly higher in samples from sites with BOP and PD ≥4mm (p 0.020 and p 0.005, respectively). Molecular Assays: Detection of Porphyromonas gingivalis (p 0.002), Tannerella forsythia (p 0.023) and Treponema denticola (p 0.015) was significantly more frequent at sites with BOP and PD ≥4mm. Conclusions: The patients living with HIV/AIDS under HAART studied here had low prevalence of clinical periodontal disease signs. However, significant detection of P. gingivalis, T. denticola, and T. forsythia in periodontal active sites, and the involvement of these microorganisms as potential HIV reactivators, show the importance of creating awareness among dental health professionals of the need for close dental and periodontal monitoring in HIV patients.

RESUMEN El objetivo de este estudio fue describir el perfil microbiológico del biofilm subgingival de los pacientes con VIH bajo tratamiento antirretroviral de alta actividad (TARGA). El estudio comprendió a 32 pacientes VIH seropositivos con enfermedad periodontal (EP) que se encontraran en tratamiento con TARGA por más de 6 meses. Los antecedentes médicos de los pacientes se obtuvieron de las historias clínicas. El examen clínico instrumental (profun didad de sondaje (PS), nivel de inserción clínico (NIC) y sangrado al sondaje (SS)) fue realizado con instrumental odontológico estándar por un investigador calibrado. De este modo, se evaluaron un total de 4.765 sitios periodontales de los cuales 125 fueron estudiados microbiológicamente. Las muestras de biope lícula subgingival se obtuvieron empleando conos de papel estéril. Las muestras se emplearon en estudios microbiológicos y moleculares por PCR de punto final. El análisis estadístico se realizó según KruskalWallis y pruebas de contrastes posthoc de DunnBonferroni. El 90,6% de la población en estudio presentó carga viral inferior a 50 copias/mm3. La prevalencia de sitios periodontales activos fue baja (1%). Los recuentos de bacterias anaerobias estrictas pigmentadas de negro y fusiformes fueron significativamente más altos en muestras de sitios periodontales con SS positivo y PS ≥4 mm (p 0.020 y p 0.005). La detección molecular de Porphyromonas gingivalis (p 0.002), Tannerella forsythia (p 0.023) y Treponema denticola (p 0.015) fue significativamente mayor en los sitios con SS y PS ≥4mm. La prevalencia del 1% de enfermedad periodontal en el grupo de pacientes estudiados fue menor a la esperada, sin embargo; la detección significativa de P. gingivalis, T. denticola y T. forsythia en sitios periodontales activos y su potencial participación como agentes reactivadores del VIH, nos alerta de la importancia de crear conciencia en los profesionales de la salud (médicos y odontólogos) acerca de la necesidad de un monitoreo minucioso del estado periodontal de pacientes con características semejantes a las descriptas en la muestra poblacional estudiada.

Prevalence and Molecular Analysis of Novel Newly Identified Periodontal Pathogens in Subgingival Plaque Samples of Saudi Patients with Chronic Periodontitis

Objective: To evaluate the prevalence of novel newly identified periodontal pathogenic strains in subgingival plaque samples and relate it with bleeding on probing, probing pocket depth and age. Material and Methods: 268 chronic periodontitis patients with a mean age of 46.0 ± 6.0 years were included. The following microorganisms were evaluated: Campylobacter gracilis (Cg), Fusobacterium nucleatum (Fn), Porphyromonas gingivalis (Pg), and Tannerella forsythia (Tf). Full mouth examination was registered; the probing pocket depth and clinical attachment level were assessed at six sites per tooth. Dental subgingival plaque samples were taken in the deepest pocket per arch in the maxilla and mandible. DNA analysis was performed using DNA-strip technology. Pearson's correlation coefficient was used for statistical analysis. Results: Porphyromonas gingivalis and Tannerella forsythia were detected at high level of 80% and 82% respectively. F nucleatum revealed a rate of 94%. Bacterial load significantly increased by increasing mean probing depth of the examined sites. Pearson's coefficient was the highest for Pg (r=0.62, p=0.0001) and the lowest for Cg (r=0.08, p=0.04). The bacterial load significantly increased by increasing the number of bleeding sites; Pearson's coefficient varied from r= 0.14 for Pg (p=0.01) to r=0.39 for Tf (p=0.001). Conclusion: It was confirmed the presence of main putative periodontal pathogens detectable in Saudi periodontal subjects, also providing a comprehensive view for correlation of these putative periodontal pathogens with the increase in probing pocket depth to the presence and load of all the bacterial species.

Diagnóstico molecular de microorganismos periodontopatógenos en pacientes alcohó-licos - fumadores con periodontitis crónica de la ciudad de Loja, Ecuador / Diagnóstico molecular de microrganismos periodontopatogênicos em pacientes taba-gistas com periodontite crônica na cidade de Loja, Equador / Molecular diagnosis of microorganisms periodontopathogens in alcoholic - smoking patients with chronic periodontitis of the city of Loja, Ecuador

Los pacientes alcohólicos y fumadores presentan mayor predisposición de desarrollar enfermedad periodontal. Objetivo: Determinar la presencia de los microorganismos periodontopatógenos: Porphyromona gingivalis, Treponema denticola, Tannerella forsythia, Fusobacterium nucleatum y Aggregatibacter actinomycetemcomitans mediante la Reacción en Ca-dena de la Polimerasa (PCR), en pacientes alcohólicos fumadores con periodontitis crónica. Materiales y métodos: El universo fue de 31 pacientes de 18 y 62 años de edad de sexo masculino alcohólicos y/o fumadores en los cuales fueron excluidos aquellos que consumían algún tipo de droga, dando una muestra de 23 pacientes con bolsas periodontales ≥ 6mm del Centro de Rehabilitación de la ciudad de Loja, Ecuador. Se realizó un examen periodontal completo y la toma de muestras en dos de los sitios más profundos de cada paciente. Las variables analizadas fueron: presencia de biofilm dental, sangrado al sondaje, profundidad al sondaje y nivel de inserción clínica. Los datos fueron analizados con la prueba de Kruskal Wallis con un nivel de significancia del 5%. Resultados: La periodontitis crónica estuvo en el 52,1% de los pacientes de 18 a 30 años, siendo más susceptibles los alcohólicos de riesgo y fumadores leves. El 91,04 % de alcohóli-cos y fumadores se encuentran asociados con la presencia de biofilm dental (p = 0,028) y en el diagnóstico molecular el 41,18% de los pacientes presentan más de 3 microorganismos (p = 0,039). Conclusión: Se evidenció la presencia de los periodontopatógenos estudiados en pacientes alcohólicos fumadores con periodontitis crónica.

Alcoholic and smoking patients have a greater predisposition to develop periodontal disease. Objective: To determine the presence of microorganisms periodontopathogens: Porphyromona gingivalis, Treponema denticola, Tannerella forsythia, Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans by Polymerase Chain Reaction (PCR), in alcoho-lic-smoking patients with chronic periodontitis. Materials and methods: The group was composed of 31 patients between the ages of 18 to 62-year-old, male alcoholics and/or smokers in which were excluded those who take some kind of drug, giving a sample of 23 patients with periodontal pockets ≥ 6mm from the Center of Rehabilitation of the city of Loja, Ecuador. A full periodontal examination and the sample taking were in two of the most profound sites of each patient. The variables analyzed were: the presence of dental biofilm, bleeding on probing, probing depth and clinical attachment level. The data were analyzed with the Kruskal Wallis Test with a significance level of 5%. Results: The chronic periodontitis was in the 52.1% of the patients between the ages of 18 to 30 years old, is more susceptible in risky alcoholics-smokers and low smokers. The 91.04 % of alcoholics and smokers are associated with the presence of dental biofilm (p = 0.028) and in the molecular diagnosis the 41.18% of patients exhibit more than 3 microorganisms (p = 0.039). Conclusion: That observed presence of the periodontopathogens in alcoholic-smoking patients with chronic periodontitis.

Os pacientes alcoólicos e fumantes apresentam uma possibilidade maior de desenvolver doença peridental. Objetivo: Determinar a presença do periodontopatógenos de microorganismos Porphyromona gingivalis, Treponema denticola, Fusobacterium nucleatum e Aggregatibacter actinomycetemcomitans por meio da Reação em Cadeia de Polimerasa (PCR),em pacientes alcoólicos e fumantes com periodontite crônico. Materiais e métodos: O universo pertenceu a 31 pacientes de 18 e 62 anos de idade de sexo masculino alcoólicos e/ou fumantes nos quais foram excluídos aqueles que consumiram algum tipo de droga, tendo uma amostra de 23 pacientes com bolsas peridentais. 6mm do Centro de Reabilitação da cidade de Loja, Equador. Um exame periodontal completo e a amostragem foram realizados em dois locais profundamente localizados de cada paciente. As variáveis analisadas eram: presença de biofilm dental, sangrou ao sondaje, profundidade ao sondaje e nível de suplemento clínico. Os dados foram analisados com o teste de Kruskal Wallis com um nível de significancia de 5%. Resultados: O periodontite crônico estava em 52,1% dos pacientes de 18 para 30 anos, enquanto sendo mais suscetível os alcoólicos de risco e fumantes ligeiros. 91,04% de alcoólicos e fumantes são associados com a presença de biofilm dental (p = 0028) e na diagnose 41,18% molecular dos pacientes apresenta mais de 3 microorganismos (p = 0039). Conclusões: A presença do periodontopatógenos foi comprovada nos pacientes alcoólicos e fumantes com periodontite crônico.

Comparison of full-mouth scaling and quadrant-wise scaling in the treatment of adult chronic periodontitis

Abstract In the search for the ideal treatment of periodontal disease various non-surgical techniques should be considered. The objective of this study was to evaluate the efficacy of full-mouth scaling (FMS) by clinical and microbiological parameters. 670 individuals were evaluated with 230 subjects meeting the selection criteria and were divided into two groups; 115 subjects treated with FMS and 115 treated with weekly sessions of scaling and root planning (SRP). The patient population had a mean age of 51.67 years, with moderate chronic periodontitis. Subjects were evaluated prior to treatment (T1) and 90 days after execution of therapy (T2), with regards to: probing depth (PD), clinical attachment level (CAL), plaque index (PI), gingival index (GI), and microbial detection for the presence of Porphyromonas gingivalis (P.g.) and Prevotella intermedia (P.i.) by culture method and confirmed by biochemical tests. Subjects treated in the FMS group also rinsed with 0.12% chlorhexidine mouthwash for seven days following treatment. The results were analyzed using statistical Student's t-test and chi-square test. No statistically significant differences were observed for PD and CAL between T1 and T2 in both groups. For GI and PI significant difference was observed between the groups. For the evaluated microbial parameters was observed reduction of P.g. and P.i., but only for P.g. with a significant reduction in both groups. The full mouth scaling technique with the methodology used in this study provided improved clinical conditions and reduction of P.g. in subjects with moderate periodontitis, optimizing the time spent in the therapeutic execution.

Resumo Na busca do tratamento ideal da doença periodontal varias são técnicas não-cirúrgicas que podem ser consideradas. O objetivo deste estudo foi avaliar a eficácia da técnica de desinfecção total de boca (FMD, na sigla em Inglês) por parâmetros clínicos e microbiológicos. Foram avaliados 670 indivíduos com 230 indivíduos atendendo aos critérios de seleção e divididos em dois grupos; 115 indivíduos tratados com FMD e 115 tratados com sessões semanais de raspagem e alisamento corono radicular (SRP, na sigla em Inglês). A população avaliada tinha idade média de 51,67 anos, com periodontite crônica moderada. Os sujeitos foram avaliados antes do tratamento (T1) e 90 dias após a execução da terapia (T2), quanto à profundidade de sondagem (PS), nível de inserção clínica (NIC), índice de placa (IP), índice gengival (IG) e detecção microbiana da presença de Porphyromonas gingivalis (P.g.) e Prevotella intermedia (P.i.) por método de cultura e confirmada por testes bioquímicos. Os indivíduos tratados no grupo FMD também realizaram bochechos com clorexidina 0,12% durante sete dias após o tratamento. Os resultados foram analisados ​​utilizando o teste estatístico t de Student e o teste de qui-quadrado. Não foram observadas diferenças estatisticamente significativas para PS e NIC entre T1 e T2 em ambos os grupos. Para IG e IP observou-se diferença significativa entre os grupos. Para os parâmetros microbianos avaliados foi observada redução de P.g. e P.i., mas apenas para P.g. com uma redução significativa em ambos os grupos. A técnica FMD com a metodologia utilizada neste estudo proporcionou condições clínicas melhoradas e redução da P.g. Em indivíduos com periodontite moderada, otimizando o tempo gasto na execução terapêutica.

PAR-2 expression in the gingival crevicular fluid reflects chronic periodontitis severity

Abstract Recent studies investigating protease-activated receptor type 2 (PAR-2) suggest an association between the receptor and periodontal inflammation. It is known that gingipain, a bacterial protease secreted by the important periodontopathogen Porphyromonas gingivalis can activate PAR-2. Previous studies by our group found that PAR-2 is overexpressed in the gingival crevicular fluid (GCF) of patients with moderate chronic periodontitis (MP). The present study aimed at evaluating whether PAR-2 expression is associated with chronic periodontitis severity. GCF samples and clinical parameters, including plaque and bleeding on probing indices, probing pocket depth and clinical attachment level, were collected from the control group (n = 19) at baseline, and from MP patients (n = 19) and severe chronic periodontitis (SP) (n = 19) patients before and 6 weeks after periodontal non-surgical treatment. PAR-2 and gingipain messenger RNA (mRNA) in the GCF of 4 periodontal sites per patient were evaluated by Reverse Transcription Polymerase Chain Reaction (RT-qPCR). PAR-2 and gingipain expressions were greater in periodontitis patients than in control group patients. In addition, the SP group presented increased PAR-2 and gingipain mRNA levels, compared with the MP group. Furthermore, periodontal treatment significantly reduced (p <0.05) PAR-2 expression in patients with periodontitis. In conclusion, PAR-2 is associated with chronic periodontitis severity and with gingipain levels in the periodontal pocket, thus suggesting that PAR-2 expression in the GCF reflects the severity of destruction during periodontal infection.

Porphyromonas gingivalis, patógeno de relevancia en la enfermedad periodontal / Porphyromonas gingivalis, pathogen of interest in periodontal disease

Porphyromonas gingivalis (P.Gingivalis) es un microorganismo comprometido en el inicio y progresión de la enfermedad periodontal crónica y agresiva, y es considerado su principal agente etiológico. Esta bacteria cuenta con una serie de factores de virulencia que le permiten, iniciar el proceso infeccioso, perpetuar la infección y también transformar la placa dental benigna en una comunidad microbiana patógena. Estudiar sus factores de virulencia y su capacidad de modular la respuesta inmunológica del huésped es muy importante para comprender el papel de este patógeno en el desarrollo y establecimiento de la enfermedad. Esta revisión proporciona una visiónactual sobre los factores de virulencia y su impacto sobre la respuesta inmunológica en relación con la patogénesis de la enfermedad periodontal.

Posible papel de Porphyromonas gingivalis en el desarrollo de la artritis reumatoide / Possible role of Porphyromonas gingivalis in the develpment of rheumatoid arthritis

Varios estudios han sugerido una asociación entre la periodontitissevera, la prevalencia de la bacteria Porphyromonas gingivalis y el desarrollo de artritis reumatoide. Como fundamento de esta relación, se ha observado que esta bacteria secreta una enzima, peptidil-arginina deiminasa, que es capaz de citrulinar proteínas del hospedero y así favorecer una respuesta autoinmune. Sin embargo, debido a la heterogeneidad de diseños experimentales, selección de pacientes y valoración de los desenlaces, los resultados no han mostrado la reproducibilidad deseada. Asimismo, observaciones recientes apuntan a que la actividad enzimática podría ser generada por otras especies bacterianas, lo que hace más compleja su relación. Sin embargo, por otro lado, algunos estudios sugieren que el tratamiento periodontal puede limitar el desarrollo de la artritis reumatoide.

Various studies have suggested a link between severe periodontitis,the prevalence of Porphyromonas gingivalis, and the development ofrheumatoid arthritis. As evidence of this relationship, P. gingivalis hasbeen found to secrete an enzyme, peptidyl arginine deiminase, which isable to citrullinate host proteins and thus help activate an autoimmuneresponse. However, due to the heterogeneity of experimental designs,patient selection, and assessment of clinical outcomes, the results havenot shown the desired reproducibility. Furthermore, recent fi ndingsindicate that the enzymatic activity may be produced by other species ofbacteria, which suggests the relationship is more complex. However, anumber of studies have shown that periodontal treatment could inhibitthe development of rheumatoid arthritis.

Microbiological effects of periodontal therapy plus azithromycin in patients with diabetes: results from a randomized clinical trial

Evidencia actual sugiere que la infección periodontal puede agravar el control de la diabetes. El objetivo de este estudiofue determinar los cambios en la frecuencia de detección de porphyromonas gingivalis, Tannerella forsythia, Treponema denticola y Aggregatibacter actinomycetemcomitans en pacientes con diabetes con el uso de terapia mecánica no quirúrgica mas azitromicina en un estudio clínico controlado aleatorizado.Ciento cinco pacientes (105) con diabetes y perio -dontitis fueron asignados aleatoriamente en tres grupos: terapiamecánica mas azitromicina, terapia mecánica mas placebo y profilaxis supragingival mas azitromicina. Un análisis periodontal completo y detección de patógenos perio dontales por medio de reacción en cadena de la polimerasa (PCR) se realizaron al inicio, 3, 6 y 9 meses después de la terapia pe -riodontal. La frecuencia de detección de Porphyromonas gingivalis, Treponema denticola y Aggregatibacter actinomycetemcomitans disminuyó a los 3 meses en todos los grupos. La frecuencia de detección de Tannerella forsythia se incrementó a los 3 meses en todos los grupos. Todos los patógenosperiodontales mostraron una frecuencia de detección similar a los 9 meses en todos los grupos. La terapia periodontal mas azitromicina no tuvo efectos adicionales sobre la frecuencia dedetección de los patógenos periodontales investigados en pacientes diabéticos...

Alteraciones morfo-cardiovasculares producidas por bacterias periodontopatógenas en modelo murino / Morphological and cardiovascular alterations produced by periodontopathic bacteria in a murine model

Diferentes autores han descrito a la enfermedad periodontal como un factor de riesgo de ateroesclerosis. Este estudio tuvo como objetivo determinar la posible relación entre las bacterias periodontopatógenas y alteraciones cardiovasculares en modelos murinos con ingesta hiperlipídica. Se utilizaron 30 ratas Wistar divididas en un grupo control (n=5) y tres grupos experimentales (n=15): Grupo I (n=5): dieta normal e infectados; Grupo II (n=5): dieta hiperlipídica y sin infectar y Grupo III (n=5): dieta hiperlipídica e infectados. La infección del surco gingival se logró previa sedación a través de hilos de sutura contaminados, colocados durante 6 días consecutivos, con Pophyromonas gingivalis (P. gingivalis) (5x10a-4 UFC) y Agregatibacter actinomycetemcomitans (A. actinomycetemcomitans) (5x10-4 UFC) obtnidas de muestras de pacientes con periodontitis crónica. A los 30 días posinfección se fijaron muestras de corazón y arteria aorta para su procesamiento. En el grupo control no se encontraron alteraciones. En el grupo I se observó en el endocardio valvular abundante exudado fibrinoleucocitario y focos hemorrágicos. En el miocardio, neovascularización y sufusiones hemorrágicos y en la pared de vasos arteriales, engrosamiento intimal. En el grupo II se observaron depósitos lipídicos en el interior de las células endoteliales, estrías grasas y pequeños ateromas. El grupo III mostró en un 100 por ciento de los casos, placas ateromatosas bien detalladas con abundante depósito de colesterol, calcificaciones, hemorragias y neovascularización. La suma de estos factores de riesgo: dieta hiperlipídica e infección con bacterias periodontopatógenas, potenciarían la expresión de alteraciones cardíacas y coronarias, incrementando la formación de ateromas

Efeito da severidade da doença periodontal sobre a expressão do Receptor Ativado por Protease do Tipo 2 (PAR-2) / Effect of the severity of periodontal disease on the expression of Protease Activated Receptor Type 2 (PAR-2)

Pesquisas recentes investigando o receptor ativado por protease do tipo 2 (PAR-2) sugerem uma associação entre este receptor e a inflamação periodontal. Além disso, é sabido que a gingipaína, protease bacteriana secretada por um importante periodontopatógeno, Porphyromonas gingivalis (Pg), tem a capacidade de ativar o PAR-2. Ademais, um estudo anterior do grupo, verificou que quanto mais profunda a bolsa periodontal, maior era a expressão do receptor PAR-2. No entanto, não é sabido se a expressão de PAR-2 é proporcional a severidade de doença periodontal e a quantidade de gingipaína expressa na bolsa periodontal. Desta forma, o presente estudo, verificou no fluido gengival a correlação entre a expressão gênica de PAR-2 (Real Time-PCR) com os parâmetros clínicos periodontais, e a expressão gênica da protease gingipaína em pacientes com periodontite crônica severa e moderada, antes e após o tratamento periodontal não-cirúrgico. A expressão de PAR-2 e da protease gingipaína foi estatisticamente maior nos pacientes do grupo periodontite crônica severa (PS) em comparação com os pacientes do grupo periodontite crônica modera (PM) e controle (C). Além disso, o tratamento periodontal levou à redução significativa (p<0.05) da expressão de PAR-2 nos pacientes com periodontite crônica moderada. Em conclusão, dentro dos limites do presente estudo, nós demonstramos que a severidade da doença periodontal e a expressão de gingipaína influenciaram a expressão de PAR-2 no fluido gengival de pacientes com periodontite crônica.

Recent studies investigating the protease-activated receptor type 2 (PAR-2) suggest an association between the receptor and periodontal inflammation. In addition, it is known that gingipain, a bacterial protease secreted by an important periodontopathogen, Porphyromonas gingivalis (Pg), has the ability to activate PAR- 2. Furthermore, a previous study from our group found that the deeper the periodontal pocket, the higher the expression of the PAR-2 receptor. However, it is not known whether the expression of PAR-2 is associated to the severity of periodontal disease and the amount of gingipain expressed in the periodontal pocket. Thus, the present study verified, in the gingival fluid, the correlation between the PAR-2 gene expression (Real Time-PCR) with the clinical periodontal parameters, and the gene expression of the gingipain protease in patients with moderate and severe chronic periodontitis before and after non-surgical periodontal treatment. PAR-2 expression and gingipain protease were statistically more expressed in patients of the severe chronic periodontitis group (PS) compared with those in the moderate chronic periodontitis group (PM) and control group (C). Furthermore, periodontal treatment led to a significant reduction (p <0.05) in the expression of PAR-2 in patients with moderate chronic periodontitis. In conclusion, within the limits of the present study, we demonstrated that the severity of periodontal disease and the expression of gingipain influenced the PAR-2 expression in the gingival fluid of patients with chronic periodontitis.

Clinical and microbiological evaluation of one-stage full-mouth disinfection: a short-term study / Avaliação clínica e microbiológica do protocolo one-stage full-mouth disinfection: estudo de curto prazo

Introduction: Patients seem to adhere better to short-term periodontal treatment schemes. Besides, time-reduced treatments are more cost-effective. However, the degree of benefits related to this type of treatment still requires additional investigations. Aim: The present short-term study evaluated clinical and microbiological outcomes, from baseline to 3-months, of chronic periodontitis subjects treated by the one-stage full-mouth disinfection protocol. Material and Method: Sixteen chronic periodontitis subjects (mean-age 49.87 ± 8.22) who met inclusion/exclusion criteria were included. A calibrated examiner measured whole-mouth plaque and gingival indices, periodontal pocket depth and clinical attachment level at baseline and at 3-months. Subgingival samples were also collected from the 5 most diseased periodontal sites to determine total bacterial load and levels of P. gingivalis and S. oralis by real time qPCR. Periodontal treatment consisted of full-mouth manual debridement plus wide intraoral use of chlorhexidine in gel and solution. Additionally, after debridement, individuals rinsed 0.12% chlorhexidine at home twice a day for the following 2 months. Data monitored were compared by paired Student-t test (p<0.05). Result: Statistical analysis revealed that, in general, one-stage full-mouth disinfection treatment provided significant clinical and microbiological improvements at 3-months. Total bacterial load showed one of the most pronounced reductions from baseline to 3-months (p=0.0001). Also, subgingival levels P. gingivalis and S. oralis reduced overtime. Conclusion: After a short period of monitoring, chronic periodontitis subjects showed clinical and microbial improvements following one-stage full-mouth disinfection treatment.

Introdução: Os pacientes parecem aderir melhor ao tratamento periodontal de curto prazo. Além disso, tratamentos de tempo curto possuem melhor custo-benefício. No entanto, os benefícios associados a este tipo de tratamento ainda requerem investigações adicionais. Objetivo: O presente estudo avaliou longitudinalmente indivíduos com periodontite crônica clínica e microbiologicamente tratados pelo protocolo one-stage full-mouth disinfection. Material e Método: Dezesseis indivíduos (49,87 ± 8,22), que se enquadraram nos critérios de inclusão/exclusão foram incluídos. Um examinador calibrado, avaliou índice de placa e gengival, profundidade de bolsa e nível clínico de inserção pré e pós terapia. Amostras subgengivais coletadas dos cinco sítios periodontais mais doentes estabeleceram a carga bacteriana total e níveis de P. gingivalis e S. oralis por qPCR. One-stage full-mouth disinfection foi realizado com instrumentos manuais associado a gel de clorexidina e solução. Após, os indivíduos utilizaram clorexidina 0,12% para bochecho, duas vezes ao dia, durante os dois meses. Dados foram comparados pelo teste pareado t de Student (p<0,05). Resultado: A análise estatística revelou que o tratamento proporcionou melhorias clínicas e microbianas significativas em três meses. A carga bacteriana total evidenciou reduções mais pronunciadas do pré para o pós-tratamento (p=0,0001). Similarmente, P. gingivalis e S. oralis mostraram redução no pós-tratamento. Conclusão: Após 3 meses de monitoramento, indivíduos com periodontite crônica apresentaram melhora clínica e microbiana com o protocolo one stage full-mouth disinfection.

Factores de virulencia de Prophyromonas gingivalis / Virulence factors of Porphyromonas gingivalis

Porphyromonas gingivalis es un microorganismo implicado en el inicio y progresión de la enfermedad periodontal crónica y agresiva, y es considerado como su principal agente etiológico. Esta bacteria cuenta con una serie de factores de virulencia importantes para su persistencia dentro de los tejidos y su resistencia al tratamiento que le permiten, primer, iniciar el proceso infeccioso a través demecanismos como la colonización, invasión y replicación dentro de las células epiteliales y fibroblastos y luego, evadir y manipular el sistema inmune del huésped y de esta forma establecer una infección crónica y por último, destruir los tejidos del huésped por medio de varias proteasas, colagenasas y enzimas. Estudiar estos factores de virulencia es muy importante para comprender el papel de este patógeno en el desarrollo y establecimiento de la enfermedad periodontal y reconcoer la importancia del control microbiológico para alcanzar el éxito en la terapia periodontal. Por otro lado, este patógeno parece estar envuelto en algunas de las manifestaciones sistémicas que genera la enfermedad periodontal no tratada. En estudio in vitro y de experimentación, ha sido vinculado con mecanismos que generan desde bajo peso al nacer, artritis reumatoidea, disfunción endotelial y peroxidación de lípidos. Estas implicaciones del patógeno periodontal lo hacen también interesante desde el punto de vista de la salud general de las comunidades.

Evaluación de la terapia mecánica periodontal en bolsas profundas: respuesta clínica y bacteriológica / Evaluation of periodontal mechanical therapy in deep pockets: Clinical and bacteriological response

Objetivos: Evaluar la composición microbiológica y los parámetros clínicos de bolsas periodontales >5 mm de profundidad al inicio, 1 semana, 3 y 12 meses post raspado y alisado radicular. Materiales y Métodos: Se tomaron registros clínicos y muestras de placa subgingival de 44 sitios de pacientes con diagnóstico de periodontitis crónica. Se identificaron por técnica de Reacción en Cadena de la Polimerasa (PCR) patógenos putativos periodontales: Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Treponema denticola (Td), Tannerella forsythia (Tf) y Prevotella intermedia (Pi). Los pacientes recibieron terapia mecánica periodontal y fueron reevaluados a los 7 días, 3 y 12 meses. Resultados: Luego del tratamiento, todos los parámetros clínicos (Placa Bacteriana, Hemorragia, Supuración, Profundidad al Sondaje y Nivel de Inserción Clínica) se redujeron significativamente y los valores obtenidos se mantuvieron hasta los 12 meses. Al inicio, las especies bacterianas prevalentes fueron Pg, presente en 66 por ciento de los sitios, Tf (55 por ciento) y Td (41 por ciento). Los sitios más profundos se relacionaron con las asociaciones Tf-Td (6.8 mm) y Tf-Td-Pi (7 mm). Post terapia, el número de sitios positivos para Td, Tf y Pg se redujo significativamente. Conclusiones: El raspado y alisado radicular mejoró significativamente los parámetros clínicos y redujo la prevalencia de los patógenos periodontales Pg, Tf y Td en bolsas periodontales profundas. Los resultados obtenidos se mantuvieron hasta los 12 meses. No se detectaron mayores pérdidas de inserción clínica en el 86 por ciento de los sitios a 3 meses y en 79 por ciento a los 12 meses. Los sitios en los que el tratamiento no fue efectivo en la eliminación de patógenos a los 12 meses desarrollaron mayores profundidades de sondaje.

Objectives: To evaluate the microbial composition and clinical parameters of periodontal pockets with probing depth >5 mm at baseline, 1 week, 3 and 12 months after scaling and root planning. Methods: Clinical parameters were measured and bacterial samples were collected from 44 sites in 11 patients with chronic periodontitis. By means of Polymerase Chain Reaction (PCR) the presence of Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Treponema denticola (Td), Tannerella forsythia (Tf) and Prevotella intermedia (Pi) was estimated. The patients received mechanical periodontal therapy and were evaluated after 1 week, 3 months and 12 months. Results: After treatment, all clinical parameters (Plaque, Bleeding on Probing, Supuration, Probing Pocket Depth and Clinical Attachment Level) were significantly reduced, and the values obtained were maintained up to the 12 months that the study lasts. At baseline, the most prevalent species were Pg, present in 66 percent of the sites, Tf (55 percent) and Td (41 percent). The deepest sites were related to the association Tf-Td (6.8 mm) and Tf-Td-Pi (7 mm). The number of positive sites for Td, Tf and Pg was significantly reduced after therapy. Conclusions: Scaling and root planning improve significantly clinical parameters as well as reduce the prevalence of periodontal pathogens Pg, Td and Tf in deep periodontal pockets. The results obtained were maintained up to 12 months. No further clinical attachment loss was found in 86 percent of the sites at 3 months and 79 percent at 12 months. The sites where the treatment failed in removing pathogens developed at 12 months greater probing pocket depths.

Increased levels of Porphyromonas gingivalis are associated with ischemic and hemorrhagic cerebrovascular disease in humans: an in vivo study

OBJECTIVE: This study investigated the role of periodontal disease in the development of stroke or cerebral infarction in patients by evaluating the clinical periodontal conditions and the subgingival levels of periodontopathogens. MATERIAL AND METHODS: Twenty patients with ischemic (I-CVA) or hemorrhagic (H-CVA) cerebrovascular episodes (test group) and 60 systemically healthy patients (control group) were evaluated for: probing depth, clinical attachment level, bleeding on probing and plaque index. Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans were both identified and quantified in subgingival plaque samples by conventional and real-time PCR, respectively. RESULTS: The test group showed a significant increase in each of the following parameters: pocket depth, clinical attachment loss, bleeding on probing, plaque index and number of missing teeth when compared to control values (p<0.05, unpaired t-test). Likewise, the test group had increased numbers of sites that were contaminated with P. gingivalis (60 percentx10 percent; p<0.001; chi-squared test) and displayed greater prevalence of periodontal disease, with an odds ratio of 48.06 (95 percent CI: 5.96-387.72; p<0.001). Notably, a positive correlation between probing depth and the levels of P. gingivalis in ischemic stroke was found (r=0.60; p=0.03; Spearman's rank correlation coefficient test). A. actinomycetemcomitans DNA was not detected in any of the groups by conventional or real-time PCR. CONCLUSIONS: Stroke patients had deeper pockets, more severe attachment loss, increased bleeding on probing, increased plaque indexes, and in their pockets harbored increased levels of P. gingivalis. These findings suggest that periodontal disease is a risk factor for the development of cerebral hemorrhage or infarction. Early treatment of periodontitis may counteract the development of cerebrovascular episodes.

Relación entre la Enfermedad Periodontal (EP) y la Enfermedad Cardiovascular (ECV): Una revisión de tema / Relationship between periodontal disease (PD) and Cardiovascular Disease (CVD): a review of topic

Several clinical studies had investigated the possible relationship between periodontal disease (PD) and cardiovascular disease (CVD). Some studies prove an epidemiological association between PD and CVD, that from the biological viewpoint may be linked to both chronic inflammation triggered by microbes and persistent microbial infection by periodontal pathogens. In that sense, individuals presenting deep periodontal pockets without periodontal treatment, increased number of missing teeth and other periodontal markers like clinical attachment level and bleeding on probing, seem to be important risk factors markers for cardiovascular disease. Both, EP and cerebrovascular diseases are frequent, and those association is important in Public Health setting. EP and CVD share risk factors such as age, tobacco, stress, socioeconomic status and fat metabolism, so that the chances of introducing confusion into the studies is high. Few population studies have examined the effect of periodontal treatment on the risk of developing cardiovascular events (either primary or secondary prevention), however, it must be noted that the evidence in human studies suggest a possible beneficial effect of periodontal treatment of cardiovascular function index. In this paper, we review the literature that enables the general dentist and specialist an approach to this problem and better understanding of this phenomenon to patient benefit...(AU)

Varios estudios clínicos han investigado la posible relación entre la enfermedad periodontal (EP) y la enfermedad cardiovascular (ECV). Algunos demuestran una asociación epidemiológica entre EP y ECV que desde el punto de vista biológico puede estar ligado con inflamación crónica e infección microbiana persistente con la presencia de patógenos periodontales. En ese sentido para los individuos el presentar las bolsas periodontales profundas sin tratamiento periodontal, el número de dientes perdidos y otros marcadores periodontales como el nivel de inserción clínica y el sangrado al sondeo, parecen ser factores de riesgo importantes para las enfermedades cardiovasculares. Ambas enfermedades EP y ECV son frecuentes, y su asociación es muy importante en salud pública, porque comparten factores de riesgo, tales como la edad, tabaco, stress, estatus socioeconómico y metabolismo de las grasas, por lo que las posibilidades de introducir confusión en los estudios son elevadas. Muy pocos estudios poblacionales han estudiado el efecto del tratamiento periodontal en el riesgo de desarrollar eventos cardiovasculares (ya sea en la prevención primaria o secundaria), sin embargo, hay que señalar que es la evidencia en estudios con seres humanos sugieren un posible efecto beneficioso del tratamiento periodontal de los índices de función cardiovascular. En este trabajo, se hace una revisión de la literatura que permita al odontólogo general y especializado, un acercamiento a esta problemática y una mayor comprensión del fenómeno para el beneficio de los pacientes...(AU)

Genótipos fimA de porphyromonas gingivalis e periodontite / Porphyromonas gingivalis fimA genotypes and periodontitis

Porphyromonas gingivalis é um dos principais patógenos envolvidos com o início e progressão da doença periodontal. Este microorganismo exibe uma diversidade genotípica e fenotípica que pode resultar em diferenças na capacidade dos clones induzirem destruição periodontal. Variações no potencial patogênico dos distintos genótipos do gene fimA de P. gingivalis foram relacionadas a diferentes doenças periodontais em diversos estudos. Esta revisão teve o objetivo de analisar criticamente os estudos que relacionaram as condições periodontais e os diferentes genótipos fimA de P. gingivalis. Foram incluídas publicações na língua inglesa de estudos clínicos em humanos, que avaliaram a relação entre as condições periodontais e os diferentes genótipos fimA de P. gingivalis. Doze estudos foram considerados válidos para a realização desta revisão. Pôde-se concluir que os genótipos fimA II e IV de P. gingivalis encontram-se frequentemente associados à periodontite crônica

Porphyromonas gingivalis is a major pathogen involved in the initiation and progression of periodontal disease. This microorganism exhibits a genotypic and phenotypic diversity which may result in differences in the ability to induce periodontal destruction of the clones. Variations in the pathogenic potential of different genotypes of fimA gene of P. gingivalis were related to different periodontal diseases in several studies. This systematic review aimed to critically analyze the studies that have linked periodontal conditions and the different fimA genotypes of P. gingivalis. We included English language publications of human clinical studies that evaluated the relationship between periodontal conditions and the different fimA genotypes of P. gingivalis. Twelve studies were considered valid for the completion of this review. It was concluded that fimA genotypes II and IV of P. gingivalis are often associated with chronic periodontitis.

Presença de Aggregatibacter actinomycetemcomitans em associação a Porphyromonas gingivalis e Prevotella intermedia em pacientes periodontais / Presence of Aggregatibacter actinomycetemcomitans in association with Porphyromonas gingivalis and Prevotella intermedia in periodontal patients

Introdução: O conhecimento das inter-relações bacterianas e sua associação com a condição clínica periodontal podem auxiliar o manejo terapêutico. Objetivos: Avaliar a distribuição de Aggregatibacter actinomycetemcomitans em relação a duas espécies bacterianas membros dos complexos Vermelho e Laranja, e analisar a relação entre a combinação de cepas leucotóxicas de A.actinomycetemcomitans, Porphyromonas gingivalis e Prevotella intermedia com a manifestação clínica da doença. Material e método: Incluíram-se 50 indivíduos (38,38 ± 10,15 anos), de ambos os gêneros, com periodontite crônica (n = 25) ou agressiva (n = 25). De cada indivíduo, amostras subgengivais foram coletadas de quatro dentes em quadrantes diferentes, com profundidade de sondagem (PS) maior ou igual a 6 mm e nível clínico de inserção (NCI) maior ou igual a 4 mm. Os parâmetros avaliados foram: clínicos - PS, NCI, Índice de Placa (IP-0/1) e Índice Gengival (IG-0/1) - e microbiológicos, com detecção de A. actinomycetemcomitans (cepas de máxima e mínima leucotoxicidades), P. gingivalis e P. intermedia por reação em cadeia da polimerase. Os dados foram estatisticamente analisados usando-se testes t-Student e Qui-quadrado (p <0,05). Resultado: Apresentaram P. gingivalis 90% dos indivíduos, P. intermedia, 82%, e A. actinomycetemcomitans, 28%. A. actinomycetemcomitans foi mais frequente em indivíduos jovens ou com periodontite agressiva. Apenas IG apresentou maiores frequências médias (p = 0,019) quando da presença simultânea de cepas A. actinomycetemcomitans de máxima leucotoxicidade, P. gingivalis e P. intermedia. Conclusão: Indivíduos com periodontite agressiva exibiram maiores frequências de A. actinomycetemcomitans. A presença de P. gingivalis e/ou P. intermedia não potencializou a patogenicidade das cepas de A. actinomycetemcomitans de máxima leucotoxicidade, pois tal combinação foi somente acompanhada de maior inflamação.

Introduction: Knowledge of bacterial interrelationships and its association with clinical periodontal condition may help the therapeutic management. Objective: To evaluate the distribution of Aggregatibacter actinomycetemcomitans in relation to 2 bacterial species members of Red and Orange Complexes; also to exam the relationship between the combined presence of A. actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia with clinical pattern of disease. Material and method: 50 (mean age 38.38 ± 10.15) subjects both gender were enrolled in the study, with chronic (n = 25) or aggressive (n = 25) periodontitis. For each subject subgingival samples were collected from 4 teeth in different quadrants showing probing depth (PD) is greater than or equal to 6 mm and clinical attachment level (CAL) is greater than or equal to 4 mm. The parameters evaluated were: clinical - PD, CAL, Plaque Index (PI-0/1) and Gingival Index (GI-0/1); microbiological - detection of A. actinomycetemcomitans (highly and minimally leukotoxic strains), P. gingivalis and P. intermedia by polymerase chain reaction. Data were statistically analyzed using Student t and Chi-square tests (p <0.05). Result: 90% of the subjects harbored P. gingivalis, 82% P. intermedia and 28% A. actinomycetemcomitans. A. actinomycetemcomitans was most frequent in young or aggressive periodontitis subjects. Only GI showed higher mean frequency (p = 0.019) in the combined presence of highly leukotoxic A. actinomycetemcomitans strains,P. gingivalis and P. intermedia. Conclusion: Subjects with aggressive periodontitis exhibited higher frequency of A. actinomycetemcomitans. The presence of P. gingivalis and/or P. intermedia did not improve the pathogenicity of highly leukotoxic A. actinomycetemcomitans strains due to the fact that this combination was only followed by increased inflammation.

Periodontal parameters and BANA test in patents with chronic renal failure undergoing hemodialysis

OBJECTIVES: The aim of this study was to analyze the periodontal parameters of patients with chronic renal failure. MATERIAL AND METHODS: The periodontal status of 16 Brazilian patients aged 29 to 53 (41.7±7.2) years with chronic renal failure (CRF) and another matched group of 14 healthy controls with periodontitis was assessed clinically and microbiologically. Probing pocket depth (PPD), gingival recession (GR), dental plaque index (PLI), gingival index (GI), and dental calculus index (CI) were the clinical parameters recorded for the entire dentition (at least 19 teeth), while the anaerobic periodontopathogen colonization in four sites with the highest PPD was evaluated using the BANA test ("PerioScan"; Oral B). RESULTS: The results for the CRF group and control group, respectively were: PPD: 1.77±0.32 and 2.65±0.53; GR: 0.58±0.56 and 0.51±0.36; PLI: 1.64±0.56 and 1.24±0.67; GI: 0.64±0.42 and 0.93±0.50; CI: 1.17±0.54 and 0.87±0.52. Comparison between groups using the "t" test revealed a significantly increased PPD (p<0.001) in the control group. Comparison of the other clincial parameters by the Mann-Whitney test showed differences only for PLI, which was significantly higher (p<0.05) in the CRF group. Spearman's test applied to each group showed a positive correlation among all clinical parameters, except for GR (p<0.05). None of the groups showed any correlation between GR and GI, while a significant negative correlation between GR and PPD was observed for the CRF group. The percentage of BANA-positive sites was 35.9 percent for the CRF group and 35.7 percent for the control group. The BANA test correlated positively with PPD only in the control group and with GR only in the CRF group. CONCLUSIONS: In spite of a higher PLI and dense anaerobic microbial population even in shallow PPD, patients with CRF exhibited better periodontal conditions than periodontitis patients, which is an evidence of altered response to local irritants.

The ability of the BANA test to detect different levels of P. gingivalis, T. denticola and T. forsythia

The aim of this study was to evaluate the ability of the BANA Test to detect different levels of Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia or their combinations in subgingival samples at the initial diagnosis and after periodontal therapy. Periodontal sites with probing depths between 5-7 mm and clinical attachment level between 5-10 mm, from 53 subjects with chronic periodontitis, were sampled in four periods: initial diagnosis (T0), immediately (T1), 45 (T2) and 60 days (T3) after scaling and root planing. BANA Test and Checkerboard DNA-DNA hybridization identified red complex species in the subgingival biofilm. In all experimental periods, the highest frequencies of score 2 (Checkerboard DNA-DNA hybridization) for P. gingivalis, T. denticola and T. forsythia were observed when strong enzymatic activity (BANA) was present (p < 0.01). The best agreement was observed at initial diagnosis. The BANA Test sensitivity was 95.54 percent (T0), 65.18 percent (T1), 65.22 percent (T2) and 50.26 percent (T3). The specificity values were 12.24 percent (T0), 57.38 percent (T1), 46.27 percent (T2) and 53.48 percent (T3). The BANA Test is more effective for the detection of red complex pathogens when the bacterial levels are high, i.e. in the initial diagnosis of chronic periodontitis.

Medición de cambios cuantitativos de la microbiota subgingival posterior a la remoción de placa bacteriana supragingival / Measurement of quantitative changes of the microbiota subgingival after to removal of bacterial plaque supragingival

Uno de los campos de interés en el estudio de la microbiología periodontal para muchos investigadores ha sido identificar si es posible que los microorganismos responsables del origen y progresión de la enfermedad periodontal que habitan sobre el margen gingival (supragingival) y bajo este (subgingival) tengan una relación directa que permita mantener interacciones influyentes en el crecimiento y desarrollo de las diferentes especies bacterianas que habitan en los tejidos periodontales. Por lo tanto, al remover los microorganismos que se ubican supragingivalmente sería posible encontrar cambios en el medio subgingival al no existir un intercambio entre los ambientes aerobios (supragingival) y anaerobios (subgingival) una vez desorganizada la placa bacteriana supragingival. Para demostrar esta relación se seleccionaron 7 individuos con diagnóstico de periodontitis crónica moderada y severa a los cuales se les realizó un destartraje supragingival de boca completa para lograr desorganizar la placa bacteriana supragingival. A su vez se tomaron muestras microbiológicas de los sacos periodontales más profundos de cada cuadrante de estos individuos, siendo la primera muestra tomada previo al destartraje supragingival considerada como muestra basal (día 0), luego se tomaron a las 24 horas, a los 7 y 21 días de removida la placa bacteriana supragingival. De los resultados del presente estudio pudimos concluir que al desorganizar el biofilm supragingival se observa una disminución en la cantidad total de microorganismos subgingivales, así como también disminuye de manera considerable la proporción de Porphyoromona gingivalis presente en el medio subgingival. Lo cual permitiría establecer la existencia de una relación directa y dependiente entre los microorganismos que habitan el medio supragingival y subgingival.

One of the fields of interest in the study of the microbiology periodontal for many investigators has been to identify if it is possible that the microorganisms responsible for the origin and progression of the disease periodontal that live on the margin gingival (supragingival) and under this (subgingival) they have a direct relation that allows to support influential interactions in the growth and development of the different bacterial species that they live in the tissue periodontal.Therefore having removed the microorganisms that are located supragingivalmente would be possible to find changes in the way subgingival when an exchange not to exist between the aerobic environments (supragingival) and anaerobic (subgingival) once disorganized the bacterial plate supragingival. To demonstrate this relation 7 individuals selected with diagnosis of periodontitis chronicle moderate and severe to which they there was realized a destartraje supragingival of complete mouth to achieve supragingival to disorganize the bacterial plate. In turn microbiological samples of the sacks took periodontales deeper of every quadrant of these individuals, being the first taken sample before the destartraje supragingival considered as sample basal (the 0th), then they took at to 1, 7 and 21 days of removed the bacterial plate supragingival anaerobios (subgingival) once disorganized the bacterial plate supragingival Of the results of the present study we could conclude that on having disorganized the biofilm supragingival a decrease is observed in the total quantity of microorganisms subgingivales, as well as also it diminishes in a considerable way the proportion of present Porphyoromona gingivalis in the way subgingival. Which would lead to thinking that there exists a direct and dependent relation between the microorganisms that live the way supragingival and subgingival.