Clinical and microbiological evaluation of non-surgical periodontal therapy in obese and non-obese individuals with periodontitis: a 9-month prospective longitudinal study

Abstract Objective Obesity is a chronic disease that negatively affects an individual's general and oral health. The present study aimed to compare the clinical and microbiological effects of non-surgical periodontal therapy with the full mouth disinfection (FMD) protocol on obese and non-obese individuals at 9 months post-therapy. Methodology This clinical study was first submitted and approved by the Ethics Committee. Fifty-five obese patients and 39 non-obese patients with periodontitis were evaluated. The full-mouth periodontal clinical parameters, clinical attachment level (CAL), probing depth (PD), gingival index (GI), and plaque index (PI), were monitored at baseline, 3, 6, and 9 months after periodontal treatment with full mouth disinfection (FMD) protocol. The mean count of Tannerella forsythia , Porphyromonas gingivalis , Treponema Denticola , and Aggregatibacter actinomycetemcomitans was determined by quantitative polymerase chain reaction on subgingival biofilm samples. Demographic data were assessed by Chi-square test. For clinical and microbiological parameters, two-factor repeated-measures ANOVA was used. Results In both groups, periodontal therapy using the one-stage full-mouth disinfection protocol significantly improved CAL, PD, GI, and PI (p<0.05). Obese and non-obese patients equally responded to non-surgical periodontal therapy (p>0.05). Microbial count found no major differences (p>0.05) between obese and non-obese individuals who had undergone non-surgical periodontal therapy. Conclusions Obesity did not affect the clinical and microbiological outcomes of non-surgical periodontal therapy.

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.

Microbiological and clinical effects of probiotics and antibiotics on nonsurgical treatment of chronic periodontitis: a randomized placebo- controlled trial with 9-month follow-up

ABSTRACT Objective: The aim of this double-blind, placebo-controlled and parallel- arm randomized clinical trial was to evaluate the effects of Lactobacillus rhamnosus SP1-containing probiotic sachet and azithromycin tablets as an adjunct to nonsurgical therapy in clinical parameters and in presence and levels of Tannerella forsythia, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans. Material and Methods: Forty-seven systemically healthy volunteers with chronic periodontitis were recruited and monitored clinically and microbiologically at baseline for 3, 6 and 9 months after therapy. Subgingival plaque samples were collected from four periodontal sites with clinical attachment level ≥1 mm, probing pocket depth ≥4 mm and bleeding on probing, one site in each quadrant. Samples were cultivated and processed using the PCR technique. Patients received nonsurgical therapy including scaling and root planing (SRP) and were randomly assigned to a probiotic (n=16), antibiotic (n = 16) or placebo (n = 15) group. L. rhamnosus SP1 was taken once a day for 3 months. Azithromycin 500mg was taken once a day for 5 days. Results: All groups showed improvements in clinical and microbiological parameters at all time points evaluated. Probiotic and antibiotic groups showed greater reductions in cultivable microbiota compared with baseline. The placebo group showed greater reduction in number of subjects with P. gingivalis compared with baseline. However, there were no significant differences between groups. Conclusions: The adjunctive use of L. rhamnosus SP1 sachets and azithromycin during initial therapy resulted in similar clinical and microbiological improvements compared with the placebo group.

Aggregatibacter actinomycetemcomitans en periodontitis crónica en individuos de Corrientes, Argentina: estudio preliminar / Aggregatibacter actinomycetemcomitans in chronic periodontitis of individuals from Corrientes, Argentina: a preliminary study

Objetivo: establecer la prevalencia de Aggregatibacter actinomycetemcomitans en pacientes con periodontitis crónica en distintos estadios yla distribución de serotipos, utilizando la técnica de reacción en cadena de la polimerasa. Materiales y métodos: participaron 54 sujetos, entre 35 y 65 años de edad, con diagnóstico de periodontitis crónica. La periodontitis se clasificó en leve, moderada y severa. Como grupo control se incluyeron 24 sujetos sin periodontitis, con los mismos parámetros de inclusión. Para la detección de Aggregatibacter actinomycetemcomitans se procesaron muestras de placa subgingival obtenidas con cono de papel absorbente, conservadas a -20ºC hasta su procesamiento. El ácido dexorribonucleico se extrajo por el método de bromuro de cetiltrimetilamonio y se utilizó la técnica de reacción en cadena de polimerasa para su identificación y serotipado. Resultados: los resultados de las reacciones fueron leidos por electroforesis en geles de agarosa teñidos con bromuro de etidio y la visualización fue realizada por transiluminación ultravioleta. La prevalencia de A. actinomycetemcomitans en periodontitis crónica fue del 14,91 por ciento; el serotipo más frecuente fue el b. No se obtuvieron diferencias estadísticamente significativas entre los grupos con y sin periodontitis (p=0,064), ni entre los estadios de periodontitis y el serotipo presente (p=0,2139). Conclusión: aunque los resultados coinciden con la bibliografía, sería conveniente repetir el estudio sobre una muestra mayor.

Porphyromonas gingivalis Fim-A genotype distribution among Colombians / Distribución del genotipo Fim-A de Porphyromonas gingivalis entre la población colombiana

Introduction: Porphyromonas gingivalis is associated with periodontitis and exhibit a wide array of virulence factors, including fimbriae which is encoded by the FimA gene representing six known genotypes. Objetive: To identify FimA genotypes of P. gingivalis in subjects from Cali-Colombia, including the co-infection with Aggregatibacter actinomycetemcomitans, Treponema denticola, and Tannerella forsythia. Methods: Subgingival samples were collected from 151 people exhibiting diverse periodontal condition. The occurrence of P. gingivalis, FimA genotypes and other bacteria was determined by PCR. Results: Porphyromonas gingivalis was positive in 85 patients. Genotype FimA II was more prevalent without reach significant differences among study groups (54.3%), FimA IV was also prevalent in gingivitis (13.0%). A high correlation (p= 0.000) was found among P. gingivalis, T. denticola, and T. forsythia co-infection. The FimA II genotype correlated with concomitant detection of T. denticola and T. forsythia. Conclusions: Porphyromonas gingivalis was high even in the healthy group at the study population. A trend toward a greater frequency of FimA II genotype in patients with moderate and severe periodontitis was determined. The FimA II genotype was also associated with increased pocket depth, greater loss of attachment level, and patients co-infected with T. denticola and T. forsythia.

Introducción: Porphyromonas gingivalis es una bacteria asociada con la periodontitis. Expresa una amplia gama de factores de virulencia, incluyendo las fimbrias, las cuales están codificadas por el gen FimA que representa seis genotipos conocidos. Objetivo: Identificar los genotipos de FimA de P. gingivalis en pacientes de Cali - Colombia, incluyendo la co -infección con Aggregatibacter actinomycetemcomitans, Treponema denticola y Tannerella forsythia. Métodos: Se obtuvieron muestras subgingivales de 151 individuos con diferentes diagnósticos periodontales. La ocurrencia de P. gingivalis, los genotipos de FimA y otras bacterias se determinó por PCR. Resultados: Porphyromonas gingivalis fue positiva en 85 pacientes. El genotipo FimA II fue más prevalente, pero no hubo diferencias significativas entre los grupos de estudio (54.3%), FimA IV fue el más frecuente en la gingivitis (13.0%). Una alta correlación (p= 0.000) se encontró entre P. gingivalis , T. denticola y T. forsythia. El genotipo FimA II estuvo correlacionado con la detección de T. denticola y T. forsythia. Conclusiones: Porphyromonas gingivalis tuvo una alta frecuencia incluso en el grupo de individuos sanos. Se encontró una tendencia hacia una mayor frecuencia de FimA II en pacientes con periodontitis moderada y severa. El genotipo FimA II también se asoció con una mayor profundidad de la bolsa, una mayor pérdida de nivel de inserción, y con los pacientes en los que se detectó co-infección con T. denticola y T. forsythia.

Subgingivally applied minocycline microgranules in subjects with chronic periodontitis: a randomized clinical and microbiological trial / Microgránulos de minociclina subgingival en sujetos con periodontitis crónica. Estudio clínico y microbiológico aleatorizado

El objetivo de este estudio fue evaluar el efecto clínico y microbiológico de microgránulos de Minociclina, colocados subgingivalmente como coadyuvante del raspaje y alisado radicular en pacientes con Periodontitis crónica severa. Participaron 26 sujetos voluntarios con Periodontitis crónica, no fumadores. Se seleccionaron 4 sitios contralaterales con Sangrado al Sondaje (SS) y Profundidad al Sondaje (PS) > 6 mm. Condición Basal (CB): se registró, PS y Nivel de Inserción (NI). Se determinó mdiante PCR la presencia de Porphyromonas gingivalis (Pg), Tannerella forsythia (Tf), Treponema denticola (Td) y Aggregatibacter actinomycetemcomitans (Aa). Un lado de la boca fue aleatoriamente asignado al tratamiento experimental: grupo T, el otro al tratamiento control: grupo C. Al día 30 y 90 se repitieron los exámenes clínicos y microbiológicos. Día 30: el SS se redujo al 81 por ciento en el grupo C y al 12 por ciento en el grupo T (p<0,05). Estas diferencias se mantuvieron al día 90 (C: 58 por ciento, T: 8 por ciento) (p<0.05). Día 30 y 90: hubo disminución de la PS en ambos grupos, siendo significativamente mayor en el grupo T (p<0,05). En ambos grupos hubo disminución significativa del NI (p<0,05), no hubo diferencias entre los grupos al día 30 y sí al día 90. A los 30 y 90 días en ambos grupos se redujo la prevalencia para Pg, Tf ,Td y Aa. A los 30 y 90 días la reducción de sitios con Pg fue mayor en el grupo T (p=0,002). A los 90 días Td disminuyó en el grupo T y aumentó en el grupo C (p=0,023). No se observaron efectos adversos. Los resultados mostraron que la aplicación subgingival de microgránulos de minociclina adjunta al raspaje y alisado radicular produjo una reducción mayor del SS, la PS y el NI que el raspaje y alisado solo, aumentó la probabilidad de suprimir Pg y retardó la recolonización con Td.

The aim of this study was to evaluate clinical and microbiological effects of subgingival minocycline microgranules when used as an adjunct to scaling and root planing in subjects with Chronic periodontitis. Twenty-six non-smoker volunteers participated in the study. Four opposite sites, clinically standardized, with bleeding on probing (BOP) and pocket depth (PD) ≥ 6 mm were selected. Baseline BOP, PD and Clinical attachment level (CAL) were measured and microbiological samples were collected from the study sites and analyzed using PCR. Porphyromonas gingivalis (Pg) Tannerella forsythia (Tf), Treponema denticola (Td) and Aggregatibacter actinomycetemcomitans (Aa) were detected. One side of the mouth was randomly allocated to the experimental treatment: scaling and root planing plus minocycline microgranules (Test group=T) and the other side of the mouth toscaling and root planing alone (Control group=C). At days 30 and 90, clinical and microbiological examination was repeated. After 30 days BOP was reduced to 81% in C and to 12% in T and at day 90 to 58% in C and to 8% in T (p<0.05). PD wassignificantly reduced in both groups (C: 4.8mm, T: 4.2mm) favoring T at days 30 and 90 (p<0.05). CAL reduction at day 30 showed no difference between groups. At day 90, CAL reduction was higher in T (p<0.05). At days 30 and 90 Pg, Tf, Td and Aa was reduced in both groups. Pg reduction wassignificantly greater in group T. At day 90 frequency of sites with Td decreased in T and increased in C (p<0.05). No adverse effect was observed. This study showed that minocycline microgranules adjunct to scaling and root planing resulted in grater reduction of BOP and PD, higher CAL gain, increased probability of Pg suppression and retarded recolonization of Td than root instrumentation alone.

Association between periodontal condition and subgingival microbiota in women during pregnancy: a longitudinal study

Objectivo In this study, the gingival conditions and the quantitative detection for Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis and Prevotella intermedia in pregnant women were determined. Material and Methods Quantitative determinations of periodontal bacteria by using a SyBr green system in women during pregnancy were performed. Women at the 2nd and 3rd trimesters of pregnancy and non-pregnant women were included in this study. A. actinomycetemcomitans was observed in high numbers in women at the 2nd and 3rd trimesters of pregnancy with a significant difference (p<0.05). F. nucleatum and P. intermedia were also observed in high levels. Results and Conclusion Our results show that pregnant women are more susceptible to gingivitis, and the presence of A. actinomycetemcomitans in subgingival biofilm might be taken into account for the treatment of periodontal disease. .

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...

Leukotoxicity of Aggregatibacter actinomycetemcomitans in generalized aggressive periodontitis in Brazilians and their family members

OBJECTIVE: The purpose of this study was to examine the leukotoxin promoter types of Aggregatibacter actinomycetemcomitans clones in subjects with generalized aggressive periodontitis (GAgP) and in their family members (FM). MATERIAL AND METHODS: Thirty-five patients with GAgP (33.9±7.1 years), 33 of their FM (22.8±11.4 years), and 41 patients with chronic periodontitis (CP) (44.1±9.4 years) were clinically analyzed using the plaque index, gingival index, probing depth (PD), and clinical attachment level (CAL). Subgingival biofilm samples were collected from four interproximal periodontal sites (>PD and >CAL) of each patient. The presence of A. actinomycetemcomitans and its leukotoxic clone was confirmed by polymerase chain reaction (PCR). RESULTS: A. actinomycetemcomitans was observed in 23 (51.1%) GAgP patients and 16 (30.1%) CP patients. Thirty-seven (94.8%) patients showed minimally leukotoxic strains and 2 (5.1%) showed highly leukotoxic strains. In the FM group, 10 (30.3%) had aggressive periodontitis (AgP), 12 (36.3%) had CP, 11 (33.3%) were periodontally healthy or had gingivitis, and 12.2% were A. actinomycetemcomitans positive. Greater full mouth PD and CAL were observed in GAgP patients positive for the bacteria than those negative for it (p<;0.05), and the presence of A. actinomycetemcomitans positively correlated with GAgP (Odds ratio, 3.1; confidence interval, 1.4-7.0; p=0.009). CONCLUSIONS: The presence of A. actinomycetemcomitans was associated with the clinical condition of GAgP, with most patients exhibiting a generalized form of the disease and minimally leukotoxic clones. Most of the relatives of GAgP patients presented either CP or AgP. .

Distribution of serotype-specific genotypes of aggregatibacter actinomycetemcomitans in Brazilian patients with down syndrome with different periodontal conditions / Distribución de los serotipos genotipos específicos de aggregatibacter actinomycetemcomitans en los pacientes brasileños con síndrome de down con diferentes condiciones periodontales

This study evaluated the occurrence of highly or minimally leukotoxic strains of Aggregatibacter actinomycetemcomitans (Aa) from patients with Down syndrome and the distribution of the different serotype-specific genotypes of this microorganism. Sixty-seven patients with Down syndrome were subjected to dental, periodontal and radiographic evaluations. Samples of subgingival biofilm were collected and plated onto TSBV agar and characteristic colonies of A. actinomycetemcomitans were identified by biochemical methods. The occurrence of this bacterium was also evaluated directly in the clinical specimens by PCR. The presence of 530 bp deletion in the promoter region was also determined by PCR in order to evaluate distribution of highly or minimally leukotoxic strains. A. actinomycetemcomitans was detected in 11.1 percent by culture and 22.2 percent by PCR from periodontally healthy subjects, 100 percent of the patients with Down syndrome with aggressive periodontitis, 50 percent and 75 percent of patients with chronic periodontitis by culture and PCR respectively. Only two patients with aggressive periodontitis were colonized by highly leukotoxic Aa. Serotype-specific genotypes a and c were the most prevalent. The results suggest the role of peculiar characteristics of Aa and patients with Down syndrome in the development of periodontitis and the influence of peculiar characteristics of the population in this process.

Este estudio evaluó la presencia de cepas altamente o mínimamente toxicas de Aggregatibacter actinomycetemcomitans (Aa) de los pacientes con síndrome de Down y la distribución de los serotipos genotipos específicos de este microrganismo por cultivo y PCR. Sesenta y siete pacientes con síndrome de Down fueron sometidos a un tratamiento dental y evaluaciones clínicas. Las muestras de biofilme subgingival fueron recogidas y cultivadas en agar TSBV y colonias características de Aa fueran identificadas mediante métodos bioquímicos. La presencia de esta bacteria se evaluó también directamente en las muestras clínicas por PCR. Los aislados y las muestras clínicas también se probaron con el fin de evaluar la distribución de serotipos de genotipos específicos por PCR, mientras que la presencia de delección de 530 bp en la región promotora del gen ltxC también fue determinado por PCR con el fin de evaluar de distribución de las cepas altamente o mínimamente toxicas. Aa fue aislado en 11,1 por ciento y 22,2 por ciento por PCR de pacientes periodontalmente sanos; en todos los pacientes con síndrome de Down con periodontitis agresiva, y en 50 por ciento y 75 por ciento de los pacientes con periodontitis crónica por cultivo y PCR, respectivamente. Sólo dos pacientes con periodontitis agresiva fueron colonizados por cepas altamente tóxicas. Los serotipos y genotipos específicos a y c fueron los más frecuentes. Los resultados sugieren una asociación de las peculiares características de Aa con las características de los pacientes con síndrome de Down en el desarrollo de la periodontitis.

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.

Presencia de Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola y Aggregatibacter actinomycetemcomitans en el biofilm subgingival de pacientes diabéticos tipo 2: estudio transversal / Presence of Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola and Aggregatibacter actinomycetemcomitans in the subgingival biofilm of diabetic mellitus 2 patients: a cross sectional study

Antecedentes: La investigación de la microflora subgingival en pacientes diabéticos tipo 2 con periodontitis ha presentado resultados contradictorios. Objetivo: Determinar la presencia de Porphyromonas gingivalis, Tannerella forshytia, Treponema denticola y Aggregatibacter actinomycetemcomitans, en el biofilm subgingival de pacientes diabéticos tipo 2 y relacionarlo con el grado de control metabólico. Método: Estudio descriptivo transversal, en el cual se analizaron 23 pacientes diabéticos derivados consecutivamente del Policlínico de Especialidades de la Universidad de los Andes. Previo consentimiento informado, se realizó un examen clínico periodontal que incluyó mediciones de profundidad al sondaje, nivel de inserción clínica y sangrado gingival. Fueron clasificados según severidad de periodontitis y control metabólico de la diabetes determinado por un promedio de 3 exámenes de hemoglobina glicosilada. La detección microbiológica se realizó mediante la técnica de reacción en cadena de la polimerasa. Resultados: En el grupo de pacientes estudiados, Treponema denticola y Tannerella forsythia fueron las bacterias más prevalentes (65.2 por ciento), seguida por Porphyromonas gingivalis (17.3 por ciento) y Aggregatibacter actinomycetemcomitans (13 por ciento). Los pacientes con peor control glicémico tuvieron una mayor presencia de Treponema denticola, Tannerella forsythia, Porphyromonas gingivalis y Agreggatibacter actinomycetemcomitans y un aumento en el índice de sangrado al sondaje. Conclusiones: En el grupo de pacientes diabéticos estudiado, las bacterias más prevalentes fueron Treponema denticola y Tannerella forsythia. Los pacientes diabéticos tipo 2 con moderado y mal control glicémico presentaron mayor presencia de los microorganismos estudiados, comparado con los grupos con mejores niveles de control glicémico.

Background: The investigation of subgingival microflora in type 2 diabetic patients with periodontitis presented conflicting results. Aim: To determine the presence of Porphyromonas gingivalis, Tannerella forshytia, Treponema denticola and Aggregatibacter actinomycetemcomitans in subgingival biofilm of patients with diabetes type 2 and to relate it to the degree of metabolic control. Method: A descriptive study, which analyzed 23 diabetic patients consecutively referred from the Internal Medicine Unit of Medicine Faculty at Universidad de los Andes was conducted. After obtaining an informed consent from the patients a clinical examination that included measurements of periodontal pocket depth, clinical attachment level and gingival bleeding was performed. The patients were classified according to the severity of periodontitis and metabolic control of diabetes as determined by an average of 3 of glycosylated haemoglobin tests. Microbial technique was performed by chain reaction of polymerase. Results: In the group of patients examined the most prevalent bacteria were, Treponema denticola and Tannerella forsythia (65.2 percent), followed by Porphyromonas gingivalis (17.3 percent) and Aggregatibacter actinomycetemcomitans (13 percent). Patients with poor glycemic control had a greater presence of Treponema denticola, Tannerella forsythia, Porphyromonas gingivalis and Agreggatibacter actinomycetemcomitans and an increase in the rate of bleeding on probing. Conclusions: In the group of diabetic patients studied, the most prevalent bacteria were Treponema denticola and Tannerella forsythia. Type 2 diabetic patients with moderate and poor glycemic control had a higher presence of these microorganisms, compared to groups with higher levels of glycemic control.

Co-detección de patógenos periodontales en pacientes chilenos con periodontitis crónica / Co-detection of periodontal pathogens in Chilean patients with chronic periodontitis

Las enfermedades periodontales asociadas a la placa dental son un importante problema de salud pública. La etiología de estas patologías es de origen multifactorial e involucra factores del hospedero, medio ambiente y de carácter infeccioso asociados a bacterias embebidas en la placa dental. Las principales bacterias asociadas a la periodontitis crónica son Porphyromonas gingivalis, Treponema denticola y Tannerella forsythia, mientras que Aggregatibacter actinomycetemcomitans se ha asociado principalmente a la periodontitis agresiva. Otro microorganismo clave en el desarrollo de la enfermedad es Fusobacterium nucleatum, el cual tiene la capacidad de co-agregarse con los patógenos periodontales y así facilitar su colonización. Para demostrar la prevalencia de estas bacterias, la co-detección y la asociación entre pacientes chilenos fumadores y no fumadores, se analizaron 67 muestras mediante PCR convencional. Los resultados mostraron que un 90 por ciento de las muestras fueron positivas para F. nucleatum, siendo la bacteria más detectada. Al analizar la co-detección entre las distintas bacterias se observa que F nucleatum está presente en más de un 80 por ciento de los casos cuando se detecta cualquiera de las cuatro bacterias restantes, mientas que A. actinomycetemcomitans se detecta en no más de un 20 por ciento al amplificar cualquiera de las bacterias restantes. Por otra parte, los resultados por género indican que existen diferencias significativas en la detección de T. forsythia, F. nucleatum y A. actinomycetemcomitans. Al considerar el factor de tabaquismo se observó que ninguna de las muestras de pacientes fumadores resultó ser positiva para A. actinomycetemcomitans.

Periodontal disease associated with dental plaque is a major public health problem. The etiology of these disorders is multifactorial involved seeing host factors, environmental factors and infectious nature associated with the presence of bacteria belonging to the plaque. The main bacteria associated with chronic periodontitis are Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia, while Aggregatibacter actinomycetemcomitans is mainly associated with aggressive periodontitis. Another key organism in the development of the disease is Fusobacterium nucleatum, which has the ability to co-aggregate with other periodontal pathogen. To demonstrate the prevalence of these bacteria and the association between Chilean smokers and nonsmokers patients, 67 samples were analyzed by conventional PCR. The results showed that 90 percent of the samples were positive for F nucleatum being the most commonly detected bacteria. By analyzing the co-detection between different bacteria shows that F nucleatum is present in more than 80 percent of cases when it detects any of the four remaining bacteria, which lie A. actinomycetemcomitans was detected in no more than 20 percent by amplifying any remaining bacteria. Moreover the results by gender indicate that significant differences exist in the detection of T forsythia, F. nucleatum and A. actinomycetemcomitans. In considering the factor of smoking was observed that none of the samples from smokers was found to be positive for A. actinomycetemcomitans.

Occurrence of Aggregatibacter actinomycetemcomitans in Brazilians with chronic periodontitis

Studies have suggested that A. actinomycetemcomitans is involved in the aetiology of aggressive periodontitis as well as in chronic periodontitis. This study was aimed at elucidating the occurrence of A. actinomycetemcomitans in a Brazilian population with chronic periodontitis. A total of 555 (mean age 33.04 ± 12.45) individuals, living in two large areas of the São Paulo State, namely "Baixada Santista" and "Vale do Paraíba", and diagnosed with mild [180 (mean age 29.59 ± 10.94)], moderate [241 (mean age 31.18 ± 11.45)] or severe [134 (mean age 33.04 ± 12.45)] chronic periodontitis were enrolled in this survey. Clinical exams including measurements of Probing Depth, Clinical Attachment Loss, Plaque and Gingival indices and subgingival microbiological assessments were performed at all population. The genomic DNA of A. actinomycetemcomitans was identified by Polymerase Chain Reaction from periodontal pocket samples. The occurrence of A. actinomycetemcomitans among chronic periodontitis subjects as well as its association with age and gender were statistically analysed using the Chi-square and Odds Ratio tests. The significance of differences was established at 5 percent (p < 0.05). A. actinomycetemcomitans was detected in 102 (18.37 percent) individuals: 29 (16.11 percent) mild; 42 (17.42 percent) moderate; and 31 (23.13 percent) severe chronic periodontitis with no statistical difference among groups. A higher occurrence of the searched bacterium was found both in the youngest group (p < 0.05) as well as in the female group (p < 0.05). This study elucidated that A. actinomycetemcomitans harbored subgingival pockets of our target group of chronic periodontitis subjects and that this bacterium seems to be inversely related to age, but related to the female gender.

Toothbrush contamination in family members

Objective: The purpose of this study was to determine the bacterial contamination of toothbrushes in family members. Materials and Methods: One hundred and two healthy subjects were included in this descriptive study. Every individual was examined clinically and microbiologically using the CPITN index and collecting subgingival plaque samples. Each participant received a toothbrush for home use and after one month they returned it to the investigators. All toothbrushes were cultured to determine the presence of periodontopathic bacteria and enteric rods. Wilkoxon signed rank test and t student test (P d"0.05) were used to compare differences in the subgingival microbiota and toothbrush contamination and CPITN index among family members. Results: A high proportion of toothbrushes resulted highly contaminated with enteric rods (P d"0.001) compared to the subgingival environment where periodontopathic bacteria were more prevalent. The most frequent microorganisms found in toothbrushes used by parents and children for one month were Enterobacteriaceae, Pseudomonadaceae species (>50 percent) and Fusobacterium spp (30 percent). Conclusions: High levels of enteric rods were commonly detected in toothbrushes used for 1 month among members of the families. These opportunistic organisms may have an important role in oral infections including gingivitis and periodontitis. Monthly replacement or disinfection of the toothbrush can reduce the risk of bacterial transmission/translocation and thus diminish the incidence of biofilm associated oral diseases.

Occurrence of Aggregatibacter actinomycetemcomitans in Brazilian indians from Umutina Reservation, Mato Grosso, Brazil

Aggregatibacter actinomycetemcomitans is associated with periodontal disease, especially localized aggressive periodontitis, produces a potent leukotoxin and its distribution is influenced by ethnic characteristics of the population. Objective: Using culture and polymerase chain reaction (PCR) techniques, this study evaluated the occurrence of this microorganism and the distribution of leukotoxic strains isolated from Indians belonging to the Umutima Reservation, Mato Grosso, Brazil. MATERIAL AND METHODS: Forty-eight native Brazilians with gingivitis and 38 with chronic periodontitis, belonging to Umutina, Paresi, Bororo, Bakairi, Kayabi, Irantxe, Nambikwara and Terena ethnicities, were studied. Subgingival, supragingival and saliva samples of each patient were collected and transferred to VMGA III medium and to ultra pure Milli Q water. Bacteria were grown on TSBV agar and incubated in anaerobiosis (90 percent N2 + 10 percent CO2) at 37ºC for 72 h. The presence of the ltx promoter was determined by PCR, and a 530 bp deletion in the promoter was evaluated by using specific primers. RESULTS: A. actinomycetemcomitans was isolated from 8.33 percent of saliva, supragingival and subgingival samples from patients with gingivitis and from 18.42 percent of saliva and supragingival biofilm, and 26.32 percent subgingival biofilm from patients with chronic periodontitis. By PCR, the bacterial DNA was detected in 8.33 percent of saliva, supragingival and subgingival biofilms from patients with gingivitis and from 23.68 percent of saliva, 28.95 percent supragingival biofilm and 34.21 percent subgingival biofilm from patients with periodontitis. All strains were grouped as non-JP2 clones based on the absence of deletion in the leukotoxin promoter. Differences among the microbial and clinical parameters in patients were analyzed by using the Mann-Whitney, Chi-square or Fisher's exact tests. CONCLUSIONS: The present results suggest that A. actinomycetemcomitans ...

Preliminary results of the use an ultrasonic equipment in the treatment of moderate periodontitis: clinical and microbiological real time polimerase chain reacion-based (PCR) study

OBJETIVE: This preliminary study aimed to evaluate clinically and microbiologically the treatment of patients with light to moderate periodontal disease by using an ultrasonic device. MATERIAL AND METHODS: Sixteen adults with moderate periodontitis were studied. A Real-time PCR protocol was optimised using specific primers for the three microorganisms. Clinical and microbiological testing was performed prior and following treatment. RESULTS: Despite clinical improvement, bacteria quantification didn’t change. All the patients were positive for P. gingivalis and P. intermedia and nine for A. actinomycetemcomitans. CONCLUSION: VectorTM-system treatment showed clinical improvement. A Real-time PCR protocol is now available for periodontal diagnostics and monitorization. However, further studies with larger populations are indicated.

OBJETIVOS: Este estudo preliminar teve a finalidade de avaliar clinicamente microbiologicamente o tratamento de pacientes portadores de periodontite leve a moderada, utilizando-se um novo aparelho piezo-elétrico disponível no mercado (VectorTM system). MATERIAL E MÉTODO: Dezesseis pacientes adultos, portadores de periodontite leve a moderada, foram submetidos ao tratamento. Utilizou-se um protocolo de PCR em tempo real utilizando primers específicos para três microrganismos. Realizaram-se avaliação clínica e um estudo microbiológico simultâneo antes e após os tratamentos. RESULTADOS: A avaliação clínica foi positiva, embora não tendo ocorrido alterações na quantidade de bactérias. Todos os pacientes apresentaram amplificação específica para P. intermedia e P. gingivalis e nove pacientes para A. Actinomycetemcomitans. CONCLUSÃO: Considerando-se os resultados deste estudo preliminar, tratamento com o aparelho testado mostrou eficácia clínica. O protocolo utilizado neste estudo está agora disponível para diagnóstico periodontal e monitoração de resultados de tratamento. Entretanto, indica-se estudos clínicos e microbiológicos mais aprofundados, com amostras maiores, para conclusões mais consistentes.

Osteomyelitis of the mandible due to Aggregatibacter (Actinobacillus) actinomycetemcomitans.

Aggregatibacter (Actinobacillus) actinomycetemcomitans is a capnoic gram negative coccobacilli known to produce juvenile periodontitis. This organism was isolated in pure culture from an unusual case of osteomyelitis of the mandible. The patient was treated with tetracycline, which is the drug of choice for A. actinomycetemcomitans and the clinical response improved. From our limited review of the literature, it appears that this is the first case of osteomyelitis due to A.actinomycetemcomitans reported in India.

Distribution of biotypes and leukotoxic activity of Aggregatibacter actinomycetemcomitans isolated from Brazilian patients with chronic periodontitis / Distribuição de biótipos e atividade leucotóxica de Aggregatibacter actinomycetemcomitans isolados de pacientes brasileiros com periodontite crônica

Aggregatibacter actinomycetemcomitans is an important etiologic agent of the periodontitis and is associated with extra-oral infections. In this study, the detection of the ltxA gene as well as the ltx promoter region from leukotoxic A. actinomycetemcomitans isolated from 50 Brazilian patients with periodontitis and 50 healthy subjects was performed. The leukotoxic activity on HL-60 cells was also evaluated. Leukotoxic activity was determined using a trypan blue exclusion method. The 530 bp deletion in the promoter region was evaluated by PCR using a PRO primer pair. A. actinomycetemcomitans was detected by culture and directly from crude subgingival biofilm by PCR using specific primers. By culture, A. actinomycetemcomitans was detected in nine (18 percent) of the periodontal patients and one (2 percent) healthy subject. However, by PCR, this organism was detected in 44 percent of the periodontal patients and in 16 percent of the healthy subjects. It was verified a great discrepancy between PCR detection of the ltx operon promoter directly from crude subgingival biofilm and from bacterial DNA. Only one periodontal sample harbored highly leukotoxic A. actinomycetemcomitans. Moreover, biotype II was the most prevalent and no correlation between biotypes and leukotoxic activity was observed. The diversity of leukotoxin expression by A. actinomycetemcomitans suggests a role of this toxin in the pathogenesis of periodontal disease and other infectious diseases

Aggregatibacter actinomycetemcomitans é um importante agente etiológico da periodontite e produz infecções extra-bucais. Neste estudo, foram detectados os biótipos, o gene ltxA associado à produção de leucotoxina e o promotor ltx em A. actinomycetemcomitans de pacientes com e sem periodontite. A atividade leucotóxica sobre células HL-60 também foi avaliada. A atividade leucotóxica foi determinada através do método de exclusão do azul de tripam. A deleção de 530 bp no promotor ltx foi avaliada usando-se o par de iniciadores PRO. A. actinomycetemcomitans foi detectado por cultura e por PCR. Por cultura, A. actinomycetemcomitans foi detectado em nove pacientes com periodontite (18 por cento) e em um indivíduo sadio (2 por cento). Por PCR esse microrganismo foi detectado em 44 por cento dos pacientes com periodontite e em 16 por cento dos saudáveis. Verificou-se diferença estatística entre a detecção do promotor do operon ltx, por PCR, diretamente do biofilme subgengival e do DNA bacteriano. Somente uma amostra clínica apresentou A. actinomycetemcomitans altamente leukotóxico. O biótipo II foi o mais prevalente e não foi observada correlação biótipo-atividade leucotóxica. A expressão da leucotoxina por A. actinomycetemcomitans na doença periodontal e outras doenças infecciosas necessita ser avaliado.

Ocorrência de Aggregatibacter actinomycetemcomitans no biofilme subgengival de crianças com idade de 6 a 12 anos / Occurrence of Aggregatibacter actinomycetemcomitans in subgengival biofilm of children aged 6-12 years old

O presente estudo avaliou as condições gengivais, sócio-econômicas e a ocorrência de A. actinomycetemcomitans em 233 crianças com idade entre 6 e 1 2 anos, em Araçatuba-São Paulo. Na avaliação clínica periodontal foram utilizados os índices de placa e de sangramento a sondagem. A coleta dos espécimes clínicos de biofilme subgengival foi realizada com cones de papel absorvente, que foram introduzidos no interior dos sulcos gengivais sadios ou inflamados, durante 1 minuto, e transferidos para tubos contendo 3ml de VMGA III e para criotubos com 300µl de água ultrapura. As amostras foram cultivadas em ágar TSBV, em anaerobiose, a 37°C, por 3 dias. A presença desse microrganismo também foi avaliada através de PCR com iniciadores específicos. As condições sócio-econômicas da população estudada foram avaliadas por meio de questionário específico. A maioria das crianças estudadas apresentou gengivite. Verificou-se que a baixa renda familiar e o número de crianças na família estão estatisticamente associados com a gengivite nessas crianças, sendo que a ocorrência de A. actinomycetemcomitans não mostrou correlação com as condições gengivais ou sócio-econômicas da população alvo.

The present study evaluated gingival, socioeconomic conditions and the occurrence of Aggregatibacter actinomycetemcomitans in 233 children from 6 to 12 years-old in Araçatuba-São Paulo. In the periodontal evaluation the plaque index and bleeding on probing index were used. Subgingival biofilm samples were taken through sterile paper points, which were transfered to VMGA III or to criotubes with 300µl of ultrapure water. Microbial isolation was carried out using TSBV agar and the occurrence of A. actinomycetemcomitans was also determined by mean of PCR using specific primer pairs. The socio-economic conditions were evaluated by specific questionaire. The vast majority of studied children exihibited gingivitis and it was statistically correlated low income, family's children number and the gingival status of the patients. The occurence of A. actinomycetemcomitans did not show any correlation between gingival or socio-economic conditions and target population.