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OBJECTIVES: This systematic review aimed to evaluate the impact of periodontal therapy on systemic biomarkers of inflammation and oxidative stress in patients with type 2 diabetes mellitus (T2DM) and periodontitis. MATERIALS AND METHODS: An electronic search without restriction on dates or languages was performed in six electronic databases, protocol records and other sources until May 2024. To develop the search strategy, clinical question was formulated using the PICOD method. Eligibility criteria included randomized controlled trials on the effects of periodontitis-therapy on the inflammatory parameters of T2DM patients. Risk of bias and certainty of evidence were assessed by RoB2 and GRADE tools, respectively. The review protocol was registered in PROSPERO platform (CRD42020206295). RESULTS: Of 1,062 records screened, the authors determined that 14 studies enrolling 1223 participants proved eligible. Moderate-quality evidence suggested a positive effect of periodontitis-therapy on serum levels of c-reactive protein [0.39 (CI95%: 0.27-0.5)], even without the use of antibiotics [0.34 (CI95%: 0.22-0.46)], in T2DM patients. The significant reduction in C-reactive protein (CRP) among smokers in favor of periodontitis-therapy was greatest at six months of follow-up. CONCLUSIONS: Non-surgical periodontal therapy improved short-term biomarkers of systemic inflammation in T2DM patients, with moderate evidence of improvement in serum levels of high sensitivity-CRP. CLINICAL RELEVANCE: Systemic inflammation in T2DM patients can be reduced after non-surgical periodontal therapy, which also has the potential to reduce the risk of other important systemic outcomes, such as cardiovascular disease.

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Objective: This systematic review aimed to investigate the changes in the composition of the subgingival microbiota among subjects with normo-weight, overweight and obesity, in conditions of periodontal health and disease. Materials and Methods: The protocol for this study was designed following PRISMA guidelines. Records were identified using different search engines (PubMed/MedLine, Scopus and Web of Science). Observational studies, in human subjects diagnosed with obesity (BMI >30kg/m2) and periodontal disease (gingivitis and periodontitis), on the analysis of subgingival microbiota were selected. Eight articles were included. Results: The subgingival microbiota of 1,229 subjects (n=894 exposure group and n=335 control group) was analyzed. Periodontal pathogens were the most common bacteria detected in subjects with obesity and periodontitis (Porphyromonas gingivalis, Tannerella forsythia, Campylobacter gracilis, Eubacterium nodatum, Fusobacterium nucleatum spp. vincentii, Parvimonas micra, Prevotella intermedia, Campylobacter rectus, and Aggregatibacter actinomycetemcomitans), as along with some accessory pathogens such as: Streptococcus gordonii, and Veillonella parvula that favor the virulence of late colonizers. Conclusions: Although there are evident alterations in the composition of the subgingival microbiota in subjects with obesity and periodontitis, it is still a challenge to identify a specific pattern of microbiota in these subjects. If associations between subgingival plaque microorganisms and obesity are confirmed, microbiome analysis could be a useful tool to improve preventive measures and the management of people with obesity.

Objetivo: Esta revisión sistemática tuvo como objetivo investigar los cambios en la composición de la microbiota subgingival entre sujetos con normopeso, sobrepeso y obesidad, en condiciones de salud y enfermedad periodontal. Materiales y métodos: El protocolo de este estudio se diseñó siguiendo las directrices PRISMA. Los registros se identificaron utilizando diferentes motores de búsqueda (PubMed/MedLine, Scopus y Web of Science). Se seleccionaron estudios observacionales en sujetos humanos diagnosticados con obesidad (IMC >30kg/m2) y enfermedad periodontal (gingivitis y periodontitis), sobre el análisis de la microbiota subgingival. Se incluyeron ocho artículos. Resultados: Se analizó la microbiota subgingival de 1229 sujetos (n = 894 grupo de exposición y n = 335 grupo de control). Los patógenos periodontales fueron las bacterias más comunes detectadas en los sujetos con obesidad y periodontitis (Porphyromonas gingivalis, Tannerella forsythia, Campylobacter gracilis, Eubacterium nodatum, Fusobacterium nucleatum spp. vincentii, Parvimonas micra, Prevotella intermedia, Campylobacter rectus y Aggregatibacter actinomycetemcomitans), junto con algunos patógenos accesorios, como Streptococcus gordonii y Veillonella parvula, que favorecen la virulencia de los colonizadores tardíos. Conclusiones: Aunque existen alteraciones evidentes en la composición de la microbiota subgingival en sujetos con obesidad y periodontitis, sigue siendo un reto identificar un patrón específico de microbiota en ellos. Si se confirman las asociaciones entre los microorganismos de la placa subgingival y la obesidad, el análisis del microbioma podría ser una herramienta útil para mejorar las medidas preventivas y el manejo de las personas con obesidad.

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OBJECTIVES: Subgingival instrumentation (SI) with probiotics may be a proposal for the treatment of periodontitis (P), for patients with type 2 diabetes mellitus (T2DM). The Lactobacillus reuteri probiotic as an adjunctive therapy in the treatment of P associated with T2DM was evaluated. MATERIALS AND METHODS: Forty diabetic participants diagnosed with P (stage III and IV, grade B) were randomized into SI + Placebo (n = 20): subgingival instrumentation plus placebo lozenges and SI + Probi (n = 20): subgingival instrumentation plus probiotics. Probing depth (PD), gingival recession (GR), clinical attachment level (CAL), plaque index (PI), bleeding on probing (BoP), and PISA index were performed at baseline and 30, 90, and 180 days. Cytokine concentration in the gingival crevicular fluid, subgingival biofilm sample, and LDL and HDL subfractions were evaluated. RESULTS: In the deep pockets, PD in SI + Probi showed increased values (p = 0.02) compared to SI + Placebo at 90 days. For CAL, SI + Probi showed increased values compared to SI + Placebo, with a significant difference at 30 days (p = 0.03), 90 days (p = 0.02), and 180 days (p = 0.04). At #PD ≥ 7 mm, SI + Probi had a more frequent number of sites (p = 0.03) compared to SI + Placebo only at baseline. For the PISA, SI + Probi showed a significant difference (p = 0.04) compared to SI + Placebo at 90 days. For cytokines, SI + Probi showed higher quantification than SI + Placebo for IL-10 (p < 0.001) at 90 days, IL-12 (p = 0.010) at 90 days, IL-1ß (p = 0.035) at 90 days, and IL-8 (p = 0.003) at baseline. SI + Placebo showed higher quantification of IL-1ß (p = 0.041) compared to SI + Probi only at 30 days. There was a reduction in all microbial complexes. SI + Probi improved LDL size (246.7 nm vs 260.4 nm; p < 0.001), while large HDL subfractions were reduced aft 180 days of treatment (24.0% vs 20.3%; p = 0.022) when compared with SI + Placebo; this response was dependent of probiotics (1.0 mg/dL vs - 6.2 mg/dL; p = 0.002). CONCLUSION: Subgingival instrumentation improved the clinical periodontal parameters in patients with T2DM. The use of L. reuteri probiotics had no additional effects compared with the placebo; however, there was a positive effect on the lipoprotein subfraction. CLINICAL RELEVANCE: Scientific rationale for study: subgingival instrumentation with probiotics may be a proposal for the treatment of periodontitis (P), especially for patients with type 2 diabetes mellitus (T2DM). PRINCIPAL FINDINGS: the use of L. reuteri probiotics had no additional effects compared with the placebo; however, there was a positive effect on the lipoprotein subfraction.  Practical implications: L. reuteri as an adjunct to subgingival instrumentation may have significant therapeutic implications in dyslipidemia.

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Dehiscence in surgeries involving membranes often leads to bacterial contamination, hindering the healing process. This study assessed bacterial colonization on various membrane materials. Polydioxanone (PDO) membranes, with thicknesses of 0.5 mm and 1 mm, and a collagen membrane were examined. Packages containing polystyrene pins were crafted using these membranes, attached to 24-well plates, and exposed to oral bacteria from supra and subgingival biofilm. After a week's anaerobic incubation, biofilm formation was evaluated using the DNA-DNA hybridization test. Statistical analysis employed the Kruskal-Wallis test with Dunn's post hoc test. The biofilm on the polystyrene pins covered by the 0.5 mm PDO membrane showed a higher count of certain pathogens. The collagen membrane had a greater total biofilm count on its inner surface compared to both PDO membranes. The external collagen membrane face had a higher total biofilm count than the 0.5 mm PDO membrane. Furthermore, the 1 mm PDO membrane exhibited a greater count of specific pathogens than its 0.5 mm counterpart. In conclusion, the collagen membrane presented more biofilm and pathogens both internally and on its inner surface.

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This study aimed to compare the subgingival microbiota of subjects with and without breast cancer (BC). Patients with BC (Group 1; n= 50) and without BC (Group 2; n=50) with periodontitis (A) and without periodontitis (B). The study was conducted in two phases (P1 and P2). One biofilm sample was collected from each subject and analyzed by DNA-DNA Hybridization (Checkerboard DNA-DNA). The relative abundance of the subgingival microbiota differed between the Case and Control groups. However, some species were higher in patients in the Case than in Control subjects and differed between the groups in both phases. Composition of the subgingival microbial community according to the Socransky complex was related to periodontal disease, followed by clinical attachment of level (CAL ≥4mm), age, and tooth loss, which were found to be abundant in Cases when compared with controls. Patients with Tumor Grade II and III had a higher prevalence of tooth loss and CAL≥4mm. It was concluded that in individuals with BC, the sub-gingival microbiota exhibited atypical changes, but they developed periodontal disease.

El objetivo de este estudio fue comparar la microbiota subgingival de sujetos con y sin cáncer de mama (CM). Pacientes con CM (Grupo 1; n= 50) y sin CM (Grupo 2; n=50) con periodontitis (A) y sin periodontitis (B). El estudio se realizó en das fases (P1 y P2). Se recogió una muestra de biopelícula de cada sujeto y se analizó mediante hibridación ADN-ADN (tablero de ajedrez ADN-ADN). La abundancia relativa de la microbiota subgingival difirió entre los grupos de Caso y Control. Sin embargo, algunas especies fueron más altas en los pacientes del Caso que en los sujetos del Control y difirieron entre los grupos en ambas fases. La composición de la comunidad microbiana subgingival según el complejo de Socransky se relacionó con la enfermedad periodontal, seguida por el nivel de inserción clínica (CAL≥4mm), la edad y la pérdida de dientes, que se mostró abundante en los casos en comparación con los controles. Los pacientes con Tumor Grado II y III tuvieron mayor prevalencia de pérdida dental y CAL≥4mm. Se concluyó que en individuos con CM la microbiota subgingival presentó cambios atípicos, pero sin embargo, desarrollaron enfermedad periodontal.

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The oral cavity constitutes a unique ecosystem with highly variable ecological niches that harbor a great variety of microorganisms, including yeasts. Molecular methods are currently considered the gold standard for identifying species, although they involve limitations associated with the disruption of yeast cell walls to release the genomic DNA (gDNA) for amplification. AIM: The aim of this study was to compare the performance of different methods for extracting gDNA from Candida albicans and Candida dubliniensis, subsequently amplifying DNA by PCR. MATERIALS AND METHOD: Fifty-two isolates (16 C. albicans and 36 C. dubliniensis) were obtained from subgingival biofilm of HIV+ patients with clinical signs of periodontal disease. The study evaluated 6 gDNA extraction methods and two PCR amplification methods. Furthermore, the presence of alleles of HWP1 gene was determined in C. albicans. RESULTS: Comparisons of six methods show statistically significant differences (p <0.001) except for C. albicans in two of them. For C. dubliniensis, statistical differences were observed in all comparisons. Commercial methods were more efficient for concentrating gDNA than in-house methods, and both PCRs were effective. Ten heterozygous C. albicans isolates for this allele were positive for the HWP1-1 / HWP1-2 allele, one was homozygous for Wild Type HWP1-1 allele, and 5 were homozygous for novel/rare HWP1-2 allele. CONCLUSIONS: This study aims to provide simple, inexpensive strategies for phenotypic identification and molecular confirmation of Candida albicans and Candida dubliniensis for non-reference laboratories with low complexity and/or low budgets.

La cavidad oral constituye un ecosistema único con nichos ecológicos muy variables, capaz de albergar una gran variedad de microorganismos, incluidas las levaduras. Los métodos moleculares son considerados actualmente los métodos de identificación definitivos ya que a diferencia de los anteriores, nos brindan una correcta sensibilidad y especificidad. Sin embargo, existen limitaciones asociadas con la ruptura de las paredes celulares de estas levaduras para liberar el ADN genómico (gADN) necesario para la amplificación. OBJETIVO: El objetivo de este estudio fue comparar el rendimiento de diferentes métodos de extracción de gADN de Candida albicans y Candida dubliniensis, amplificando posteriormente por PCR. Materiales y Método: Se estudiaron 52 aislamientos, 16/52 de Candida albicans y 36/52 de Candida dubliniensis obtenidos de biofilm subgingival de pacientes VIH+ con signos clínicos de enfermedad periodontal. Se evaluaron seis métodos de extracción de gADN y la posterior amplificación se realizó por dos técnicas de PCR. Además en C. albicans se determinó la presencia de alelos para el gen HWP1. RESULTADOS: Las comparaciones de seis métodos son estadísticamente significativas (p<0,001) excepto para C. albicans en dos de ellos. Para C. dubliniensis se observaron diferencias estadísticas en todas las comparaciones. Los métodos comerciales mostraron una mayor eficiencia en la concentración de gADN que los métodos caseros y ambos fueron efectivos en las dos PCR. 10 aislados de C. albicans resultaron positivos para el alelo HWP1-1/HWP1-2, siendo heterocigotos para este alelo. Solo un aislamiento fue homocigoto para el alelo HWP1-1 de tipo salvaje y 5 eran homocigotos para el alelo HWP1-2 nuevo/raro. CONCLUSIONES: Este estudio tiene como objetivo proporcionar estrategias simples y económicas para la identificación fenotípica y confirmación molecular de Candida albicans y Candida dubliniensis para laboratorios de no referencia con baja complejidad y/o bajo presupuesto económico.

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ABSTRACT The oral cavity constitutes a unique ecosystem with highly variable ecological niches that harbor a great variety of microorganisms, including yeasts. Molecular methods are currently considered the gold standard for identifying species, although they involve limitations associated with the disruption of yeast cell walls to release the genomic DNA (gDNA) for amplification. Aim: The aim of this study was to compare the performance of different methods for extracting gDNA from Candida albicans and Candida dubliniensis, subsequently amplifying DNA by PCR. Materials and Method: Fifty-two isolates (16 C. albicans and 36 C. dubliniensis) were obtained from subgingival biofilm of HIV+ patients with clinical signs of periodontal disease. The study evaluated 6 gDNA extraction methods and two PCR amplification methods. Furthermore, the presence of alleles of HWP1 gene was determined in C. albicans. Results: Comparisons of six methods show statistically significant differences (p<0.001) except for C. albicans in two of them. For C. dubliniensis, statistical differences were observed in all comparisons. Commercial methods were more efficient for concentrating gDNA than in-house methods, and both PCRs were effective. Ten heterozygous C. albicans isolates for this allele were positive for the HWP1-1 / HWP1-2 allele, one was homozygous for Wild Type HWP1-1 allele, and 5 were homozygous for novel/rare HWP1-2 allele. Conclusions: This study aims to provide simple, inexpensive strategies for phenotypic identification and molecular confirmation of Candida albicans and Candida dubliniensis for non-reference laboratories with low complexity and/or low budgets.

RESUMEN La cavidad oral constituye un ecosistema único con nichos ecológicos muy variables, capaz de albergar una gran variedad de microorganismos, incluidas las levaduras. Los métodos moleculares son considerados actualmente los métodos de identificación definitivos ya que a diferencia de los anteriores, nos brindan una correcta sensibilidad y especificidad. Sin embargo, existen limitaciones asociadas con la ruptura de las paredes celulares de estas levaduras para liberar el ADN genómico (gADN) necesario para la amplificación. Objetivo: El objetivo de este estudio fue comparar el rendimiento de diferentes métodos de extracción de gADN de Candida albicans y Candida dubliniensis, amplificando posteriormente por PCR. Materiales y Método: Se estudiaron 52 aislamientos, 16/52 de Candida albicans y 36/52 de Candida dubliniensis obtenidos de biofilm subgingival de pacientes VIH+ con signos clínicos de enfermedad periodontal. Se evaluaron seis métodos de extracción de gADN y la posterior amplificación se realizó por dos técnicas de PCR. Además en C. albicans se determinó la presencia de alelos para el gen HWP1. Resultados: Las comparaciones de seis métodos son estadísticamente significativas (p<0,001) excepto para C. albicans en dos de ellos. Para C. dubliniensis se observaron diferencias estadísticas en todas las comparaciones. Los métodos comerciales mostraron una mayor eficiencia en la concentración de gADN que los métodos caseros y ambos fueron efectivos en las dos PCR. 10 aislados de C. albicans resultaron positivos para el alelo HWP1-1/HWP1-2, siendo heterocigotos para este alelo. Solo un aislamiento fue homocigoto para el alelo HWP1-1 de tipo salvaje y 5 eran homocigotos para el alelo HWP1-2 nuevo/raro. Conclusiones: Este estudio tiene como objetivo proporcionar estrategias simples y económicas para la identificación fenotípica y confirmación molecular de Candida albicans y Candida dubliniensis para laboratorios de no referencia con baja complejidad y/o bajo presupuesto económico.

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The success of a prosthetic treatment is closely related to the periodontal health of the individual. The aim of this article was to review and present the importance of prosthetic restorative materials on the condition of the periodontium, the changes that occur in the composition of the subgingival microbiota and the levels of inflammatory markers in gingival crevicular fluid. Articles on the influence of different prosthetic restorative materials on subgingival microbiota and proinflammatory cytokines were searched for using the keywords "prosthetic biomaterials", "fixed prosthesis", "periodontal health", "subgingival microbiota", "periodontal biomarkers" and "gingival crevicular fluid" in PubMed/Medline, Science Direct, Scopus and Google Scholar. The type of material used for prosthesis fabrication together with poor marginal and internal fit can result in changes in the composition of the subgingival microbiota, as well as increased accumulation and retention of dentobacterial plaque, thus favoring the development of periodontal disease and prosthetic treatment failure. Biological markers have helped to understand the inflammatory response of different prosthetic materials on periodontal tissues with the main purpose of improving their clinical application in patients who need them. Metal-free ceramic prostheses induce a lower inflammatory response regardless of the fabrication method; however, the use of CAD/CAM systems is recommended for their fabrication. In addition, it is presumed that metal-ceramic prostheses cause changes in the composition of the subgingival microbiota producing a more dysbiotic biofilm with a higher prevalence of periodontopathogenic bacteria, which may further favor periodontal deterioration.

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Abstract This study aimed to evaluate the effectiveness of using an ionized monocalcium phosphate and enamelin derivatives (IMP+ED) based mouthwash for the treatment of dentin hypersensitivity (DH) after scaling and root planing (SRP). 47 patients who reported DH after SRP treatment were included in this prospective cohort study. The Schiff Cold Air Sensitivity Scale (SCASS) was applied to classify their degree of pain in mild, moderate or intense at two times: after SRP (T0), and after one month of using a IMP+ED-based mouthwash (T1). The McNemar-Bowker test was used to compare the correlated proportions between both times (p<0.05). After the SRP therapy (T0), all the sample members reported pain distributed in the following manner: 12.8% were mild, 27.6% moderate, and 59.6% intense. At one month since treatment and with the use of the IMP+ED-based mouthwash (T1), the distribution of pain levels changed to 83% mild, 12.8% moderate, and 4.3% intense, this change was statistically significant (p<0.001). IMP+ED-based mouthwash produces a positive effect in reducing painful responses caused by exposure of the dentin tubules to the oral environment after SRP therapy.

Resumen El objetivo de este estudio fue evaluar la efectividad de un enjuague bucal a base de fosfato monocálcico ionizado y derivados de enamelina (FCI+DE) para el tratamiento de hipersensibilidad dentinaria (HD) posterior al tratamiento de raspado y alisado radicular (RAR). 47 pacientes que reportaron tener HD posterior al tratamiento de RAR fueron incluidos en este estudio prospectivo de cohorte. Con el fin de clasificar la HD de los pacientes en leve, moderada o intensa se utilizó la Escala de Sensiblidad al Aire Frío de Schiff (ESAFS). Los pacientes fueron evaluados después del tratamiento de RAR (T0) y posterior al uso de un enjuague bucal basado en FCI+DE (T1). Para comparar las proporciones correlacionadas se utilizó la prueba de McNemar-Bowker (p<0.05). La distribución del dolor de los pacientes posterior al tratamiento de RAR (T0) fue la siguiente: 12.8% fueron leves, 27.6% moderado, and 59.6% intenso. Un mes después del uso del enjuague buccal basado en FCI+DE (T1) la distribución en los niveles de dolor cambio a 83% leve, 12.8% moderado, and 4.3% intenso, este cambio fue estadísticamente significativo (p<0.001). El uso del enjuague bucal basado en FCI+DE produce una reducción significativa a la respuesta de dolor causada por la exposición de la dentina al ambiente oral como consecuencia del tratamiento de RAR.

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(1) Background: Probiotics can be considered a non-invasive periodontal monotherapy for the modulation of microbiota when periodontal treatment is not accessible. The aim was to evaluate the ability of Lactobacillus reuteri Prodentis as monotherapy to modulate periodontal parameters and subgingival biofilm dysbiosis. (2) Methods: A 30-year-old patient with periodontitis was followed longitudinally after one month of daily consumption of L. reuteri Prodentis (T0). Periodontal measurements and microbial identification by Checkerboard DNA−DNA hybridization of 40 bacteria were compared between baseline (T0) and 30 days (T1) or 90 days (T2), using the Kruskal−Wallis (KW) and Mann−Whitney U (MW) tests. (3) Results: Low values of pocket depth, attachment level, dental plaque, gingival erythema (GE), and suppuration were observed at T0 vs. T1, with the clinical improvement of GE (p < 0.05, MW) and the recovery of tooth 46 fistulation. T1 vs. T0 comparisons showed lower levels (Lev) or proportions (Prop) of Parvimonas micra (Lev: p < 0.05, MW; Prop: p < 0.01, MW) and Streptococcus gordonii (Prop: p < 0.05, MW), and a predominance (Lev/Prop) of Actinomyces odontolyticus and Streptococcus mitis; lower levels and proportions of P. micra, Eubacterium saburreum, Porphyromonas gingivalis, and Tannerella forsythia were observed in tooth 46 (T1/T2 vs. T0). (4) Conclusions: Under monotherapy with L. reuteri Prodentis, periodontal measurements of the patient were maintained, with selective changes in the subgingival microbiota that were proportional to the time of probiotic administration, with any additional periodontal treatment.

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The objective was to characterize and compare the subgingival microbiota in patients diagnosed according to the World Workshop on the Classification of Periodontal and Peri-Implant Diseases and Conditions 2018. For this cross-sectional study, Spanish and Colombian subjects (characterized as health/gingivitis, periodontitis in stages I-II or stages III-IV) were clinically assessed, and subgingival samples were taken and processed by culture. The comparisons among patients with periodontal status (and between countries) was made using Mann-Whitney, Kruskal-Wallis, ANOVA and chi-square tests. The final sample consisted of 167 subjects. Eikenella corrodens and Parvimonas micra were more frequently detected in health/gingivitis and Porphyromonas gingivalis in periodontitis (p < 0.05). Higher total counts were observed in Colombia (p = 0.036). In Spain, significantly higher levels of P. gingivalis and Campylobacter rectus were observed, and of Tannerella forsythia, P. micra, Prevotella intermedia, Fusobacterium nucleatum, Actinomyces odontolyticus and Capnocytophaga spp. in Colombia (p < 0.001). P. micra was more prevalent in health/gingivitis and stage I-II periodontitis in Colombia, and P. gingivalis in all periodontitis groups in Spain (p < 0.05). As conclusions, significant differences were detected in the microbiota between health/gingivitis and periodontitis, with minor differences between stages of periodontitis. Differences were also relevant between countries, with Colombia showing larger counts and variability of bacterial species.

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Cardiovascular diseases are the leading cause of morbidity and mortality worldwide. High blood pressure in particular, continues to increase throughout the global population at an increasingly fast pace. The relationship between arterial hypertension and periodontitis has been recently discussed in the context of its origins and implications. Particularly relevant is the role of the periodontal microbiome linked to persistent local and systemic inflammation, along with other risk factors and social determinants of health. The present protocol will investigate/assess the association between periodontal disease and its microbiome on the onset of hypertension, within a cohort from Mexico City. One thousand two hundred twelve participants will be studied during a 60-month period. Studies will include analysis of periodontal conditions, sampling and sequencing of the salivary and subgingival microbiome, interviews on nutritional and lifestyle habits, social determinants of health, blood pressure and anthropometric measurements. Statistical associations and several classic epidemiology and machine learning approaches will be performed to analyze the data. Implications for the generation of public policy-by early public health interventions or epidemiological surveillance approaches-and for the population empowerment-via the establishment of primary prevention recommendations, highlighting the relationship between oral and cardiovascular health-will be considered. This latter set of interventions will be supported by a carefully planned science communication and health promotion strategy. This study has been registered and approved by the Research and Ethics Committee of the School of Dentistry, Universidad Nacional Autónoma de México (CIE/0308/05/2019) and the National Institute of Genomic Medicine (CEI/2020/12). The umbrella cohort was approved by the Institutional Bioethics Committee of the National Institute of Cardiology-Ignacio Chavez (INC-ICh) under code 13-802.

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La periodontitis crónica es una patología caracterizada por la destrucción de los tejidos de soporte del diente. Existe evidencia científica de la presencia en bolsas periodontales de Pseudomonas aeruginosa, bacteria que altera la microbiota subgingival. Ha sido asociada al fracaso en el tratamiento de la periodontitis y podría constituir un riesgo para la salud general de los pacientes. El objetivo de esta investigación fue determinar la frecuencia de P. aeruginosa en bolsas periodontales de pacientes con periodontitis crónica que acudieron a la cátedra de Periodoncia de la Universidad Autónoma de Asunción. El estudio fue observacional descriptivo de corte transversal, para el mismo fueron seleccionados pacientes con periodontitis crónica que cumplían con los criterios de inclusión. Las piezas dentariasseleccionadas para la toma de muestras fueron aisladas con rollos de algodón estéril, y una vez removida la placa bacteriana supragingival, se retiró la placa subgingival de las bolsas periodontales por medio de curetas de Gracey y se introdujeron en tubos de ensayo que contenían medio de Stuart, para luego llevar las muestras hasta el laboratorio de microbiología para su análisis. Del total de 14 muestras, solo una dio positivo a P. aeruginosa representando el 7,14%. El microrganismo aislado resultó resistente a los antibióticos utilizados como coadyuvantes en el tratamiento de la periodontitis crónica,al igual que en otros trabajos encontrados en la literatura internacional. La presencia de estas bacterias dificulta el control de la enfermedad periodontal y podría significar un riesgo para desarrollar infecciones oportunistas, especialmente en los pacientes inmunodeprimidos

Chronic periodontitis is a pathology characterized by the destruction of tooth supporting tissues. There is scientific evidence of the presence in periodontal pockets of Pseudomonas aeruginosa, a bacterium that alters the subgingival microbiota and that has been associated with failure to treat periodontitis and could constitute a risk to patient general health. The objective of this research was to determine the frequency of P. aeruginosa in periodontal pockets of patients with chronic periodontitis who attend the Chair of Periodontology at the Autonomous University of Asunción. Descriptive, cross-sectional observational study, in which patients with chronic periodontitis who met the inclusion criteria were selected. The pieces selected for sampling were isolated with sterile cotton rolls, and once the supragingival bacterial plaque had been removed, the subgingival plaque was removed from the periodontal pockets by means of Gracey curettes and placed in test tubes containing Stuart medium, then the samples were taken to the microbiology lab for analysis. Of the total of 14 samples, only one was positive for P. aeruginosa, representing 7.14%. The isolated microorganism was resistant to the antibiotics used as adjuvants in the treatment of chronic periodontitis, as in other studies found in the international literature. The presence of these bacteria makes it difficult to control periodontal disease and could pose a risk for developing opportunistic infections, especially in immunosuppressed patients

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BACKGROUND AND OBJECTIVE: Human cytomegalovirus (HCMV) has been associated with periodontitis and apical periodontitis. The objective of this systematic review was to evaluate the association between HCMV and periodontitis, and apical periodontitis of endodontic origin. MATERIAL AND METHODS: A systematic review was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement guidelines and registered in the International prospective register of systematic reviews (PROSPERO). The search for potential studies was performed in MEDLINE via PubMed, Scopus, and SciELO. A quality assessment of the studies, publication bias analysis, and meta-analysis was performed. The results are presented in odds ratio with 95% confidence interval with the corresponding Forest plot. Sensitivity analysis was performed to evaluate the consistency of the results. RESULTS: Thirty-two studies were included in the quantitative and qualitative analyses. Of these, 26 were in periodontitis patients and 6 in apical periodontitis patients. Forest plot of combined studies revealed significant increased odds for periodontitis when subgingival HCMV was detected (OR 5.31; 95% CI 3.15-8.97). Sensitivity analysis based on quality of the included studies, showed consistent results. In contrast, the odds ratio for apical periodontitis when HCMV was detected from apical lesions was not statistically significant (OR 3.65; 95% CI 0.49-27.10). CONCLUSIONS: The results from the meta-analysis indicate that HCMV is significantly associated with periodontitis. In contrast, HCMV infection is not associated with apical periodontitis.

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Objetivos: A cavidade oral pode atuar como reservatório de vários patógenos de importância clínica, incluindo bacilos Gram-negativos (BGN) e Enterococcus spp. Essas espécies podem ainda aumentar em uma condição disbiótica, como ocorre nas doenças periodontais. Assim, este estudo teve como objetivo determinar a prevalência, susceptibilidade antimicrobiana e presença de fatores de virulência de BGN e enterococos isolados do biofilme subgengival de indivíduos com diferentes condições periodontais, correlacionando esses achados com parâmetros clínicos e composição da microbiota subgengival. Métodos: Na análise dos BGN, amostras de biofilme subgengival foram obtidas de indivíduos com Saúde Periodontal (SP, n=81), Gengivite (G, n=74) e Periodontite (P, n=207); para Enterococcus spp., amostras foram coletadas de 139 indivíduos com SP, 103 com G e 305 com P. As amostras foram cultivadas em meio seletivo e as colônias isoladas e identificadas por MALDI-ToF. A susceptibilidade antimicrobiana foi determinada por disco-difusão (CLSI); já os genes de virulência de enterococos e genes codificadores de ESBL e carbapenemases em BGN foram pesquisados por PCR. A produção de ESBL e carbapenemases por BGN foi avaliada pelo teste de sinergia de disco duplo e hidrólise de imipenem por espectrofotometria, respectivamente. A microbiota subgengival desses indivíduos foi determinada pelo Checkerboard. Diferenças entre os grupos foram avaliadas pelos testes de KruskalWallis, Mann-Whitney e Qui-quadrado. Resultados: BGN foram isolados em 36.2% das amostras, com maior prevalência (p<0,001) em pacientes com P (46.4%) em comparação com SP (22.2%) e G (22.9%). Pseudomonas aeruginosa (27.5%), Enterobacter cloacae (16.8%) e Enterobacter asburiae (10.7%) foram as espécies mais predominantes. Resistência/sensibilidade reduzida a ≥ 1 antimicrobiano foi encontrada em 60% dos BGN, mas apenas 4.6% eram multirresistentes. Altas taxas de resistência (>40%) foram observadas na família Enterobacteriacea para cefoxitina, cefalotina, amoxicilina- clavulanato e cefazolina. Uma única cepa de K. pneumoniae apresentou resistência/sensibilidade reduzida ao imipenem, embora o fenótipo ESBL e a detecção dos genes codificadores de beta-lactamases foram negativos. Enterococcus spp. foram isolados em 7.4% de todas as amostras, 53.7% eram E. faecalis. Essas espécies foram mais predominantes na P (9.8%) e G (7.8%) do que na SP (2.2%, p< 0,05); entretanto não houve correlação com os níveis de gravidade da P. Altas taxas de resistência/susceptibilidade reduzida foram observadas para ciprofloxacina, eritromicina e rifampicina. Os fatores de virulência mais predominantes incluíram ace, asa e esp, todos relacionados à formação de biofilme e colonização. F. nucleatum foi mais prevalente na microbiota de indivíduos enterococos +. Por outro lado, Dialister pneumosintes foi pouco detectado em indivíduos portadores de enterococos bopD+. Estreptococos orais foram prevalentes (>70%) na microbiota de pacientes que apresentavam enterococos suceptíveis à doxiciclina (p<0,05), frequentemente bopD- e esp- (p<0,01). Conclusão: Uma prevalência elevada de BGN da família Enterobacteriacea com resistência a cefalosporinas e penicilina é observada na microbiota subgengival de indivíduos com P. Enterococcus spp., principalmente E. faecalis são pouco frequentes na microbiota subgengival associada à SP, porém aumentam significativamente nas doenças periodontais. Os mesmos apresentam diversos genes de virulência compatíveis com destruição tecidual, bem como resistência a antimicrobianos de uso na clínica periodontal, o que pode limitar uma resposta terapêutica favorável. (AU)

Background/Aim: The oral cavity can act as a reservoir for several pathogens of clinic importance, including Gram-negative bacilli (GNB) and Enterococcus spp. These species may increase even more in a dysbiotic condition as seen in periodontal diseases. Thus, this study aimed to determine the prevalence, antimicrobial susceptibility and virulence factors of GNB and enterococci isolated from subgingival biofilm of individuals with different periodontal conditions, correlating these findings with clinical parameters and the composition of the subgingival microbiot. Methods: For GNB analysis, subgingival biofilm was obtained from individuals with periodontal health (PH, n=81), gingivitis (G, n=74) and periodontitis (P, n=207), whereas for enterococci isolation samples were taken from 139 patients with PH, 103 with G, and 305 with P. Samples were cultivated in selective media and isolated colonies were identified by MALDI-ToF. Antimicrobial susceptibility was determined by CLSI disk diffusion, whereas virulence genes by PCR. Production of ESBL and carbapenemases were evaluated by double disk synergy test and spectrophotometric detection of imipenem hydrolysis, respectively, and ESBL and carbapenemase encoding genes were surveyed by PCR. The subgingival microbiota was determined by checkerboard. Differences among groups were examined by Chi-square, Kruskal-Wallis or Mann-Whitney tests. Results: GNB were isolated from 36.2% of all samples, with a significantly greater prevalence (p<0.001) in P patients (46.4%) compared to PH (22.2%) and G (22.9%). Pseudomonas aeruginosa (27.5%), Enterobacter cloacae (16.8%) and Enterobacter asburiae (10.7%) were the most predominant species. Resistance/reduced sensitivity to ≥ 1 antimicrobial was found in 60% of GNB, but only 4.6% were multidrug resistant. High resistance rates (>40%) were seen in the Enterobacteriaceae family for cefoxitin, cephalotin, amoxicillin-clavulanate, and cefazolin. One strain of K. pneumoniae showed resistance/reduced sensitivity to imipenem, although the ESBL-phenotype and PCR targeting beta-lactamase encoding genes were negative. Enterococcus spp. were isolated from 7.4% of all samples; 53.7% were E. faecalis. Enterococci were more predominant in P (9.8%) and G (7.8%) samples than PH (2.2%; p<0.05), however there were no associations with distinct levels of disease severity. High rates of low susceptibility/resistance were seen for ciprofloxacin, erythromycin and rifampicin. Predominant virulence factors included ace, asa and esp, all related to colonization and biofilm formation. F. nucleatum was prevalent in the microbiota of enterococci+ individuals. In contrast, lower frequency of Dialister pneumosintes was found in patients carrying bopD+ enterococci. Oral streptococci were prevalent (>70%) in the microbiota of patients carrying enterococci susceptible to doxycycline (p<0.05), which were also frequently bopD- and esp- (p<0.01). Conclusion: A high prevalence of GNB of the Enterobacteriacea family, resistant to cephalosporins and penicillins is observed in the subgingival microbiota of patients with P, Enterococcus spp., mainly E.faecalis are not commonly detected in the healthy-related subgingival microbiota, however their frequency increases significantly in patients with periodontal diseases. These species carry several genes related to tissue destruction, as well as resistance to antimicrobials routinely used in the periodontal clinic, which may hinder a successful therapeutic response. (AU)

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BACKGROUND: Type-2-Diabetes (T2D) and Periodontitis are major inflammatory diseases. However, not much is known about the specific subgingival microbiota in Mexicans with diabetes and metabolic dysbiosis. The aim of this study was to describe the subgingival microbiota of Mexicans with T2D and the different periodontal and metabolic conditions, through "Checkerboard" DNA-DNA hybridization. METHODS: Subjects were divided into two groups-periodontal-health (PH) (PH_non-T2D; n = 59, PH_T2D; n = 14) and generalized-periodontitis (GP) (GP_non-T2D; n = 67, GP_T2D; n = 38). Obesity (BMI ≥ 30 kg/m2) and serum levels of glycated-hemoglobin (HbA1c), total-lipids, triglycerides, total-cholesterol, high-density-lipids, and low-density-lipids were measured for the T2D individuals. Subgingival microbial identification was processed for 40 species through DNA-probes. RESULTS: Subjects with T2D harbored significantly higher mean total levels (PH: p < 0.001, and GP_NS), a lower proportion of "red" complex (GP: p < 0.01), a higher proportion of "yellow" (GP; p < 0.001), and "orange" (GP; p < 0.01) complex than the non-T2D. GP_T2D individuals exhibited a greater proportion of putative-species-Campylobacter gracilis and S. constellatus (p < 0.001), and Parvimonas micra and Prevotella nigrescens (p < 0.01), than GP_non-T2D. T2D individuals with HbA1c > 8% had presented significantly higher mean pocket-depth and higher levels of G. morbillorum (p < 0.05) and those with obesity or dyslipidemia harbored higher levels, prevalence, or proportion of Streptococcus sp., Actinomyces sp., and Capnocytophaga sp. CONCLUSIONS: T2D individuals harbored a particular microbial profile different to non-T2D microbiota. Metabolic control was related to dysbiosis of microbiota-HbA1c>8% related to periodontitis and obesity or dyslipidemia with the predominance of saccharolytic bacteria, irrespective of their periodontal condition.

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Periodontitis is an inflammatory response in a susceptible host caused by complex microbiota, predominantly composed of Gram-negative anaerobic bacteria. Aiming to characterize the subgingival bacterial microbiota associated with ovine periodontitis, polymerase chain reaction (PCR) was performed in subgingival periodontal pocket samples of 14 sheep with severe periodontitis and in subgingival sulcus biofilm of 14 periodontally healthy sheep in search mainly of Gram-negative and Gram-positive microorganisms considered important periodontopathogens. The most prevalent bacteria in the sheep with periodontal lesions were Tannerella forsythia (78.6%), Treponema denticola (78.6%), Fusobacterium nucleatum (64.3%), and Porphyromonas gingivalis (50%), whereas in the healthy sheep, F. nucleatum (42.8%) was the most often detected bacterium. Statistically significant differences were observed for Campylobacter rectus, Enterococcus faecium, Prevotella nigrescens, T. forsythia, and T. denticola (p<0.05) in the sheep with periodontitis in the comparison between groups. Aggregatibacter actinomycetemcomitans, Enterococcus faecalis, and Porphyromonas gulae were not detected in any of the samples analyzed. In conclusion, C. rectus, E. faecium, P. nigrescens, T. forsythia, and T. denticola were associated with severe lesions caused by ovine periodontitis, and F. nucleatum was the most prevalent microorganism in the subgengival sulcus biofilm of healthy sheep.(AU)

Periodontite é a resposta inflamatória de um hospedeiro suscetível causada por complexa microbiota, composta predominantemente por bactérias anaeróbias Gram-negativas. Com o objetivo de caracterizar a microbiota bacteriana subgengival associada à periodontite ovina foi realizada a reação em cadeia da polimerase (PCR) de amostras de biofilme subgengival de 14 ovinos com a enfermidade e 14 ovinos periodontalmente saudáveis, com destaque para micro-organismos Gram-negativos e Gram-positivos considerados importantes periodontopatógenos. As bactérias mais prevalentes em 14 animais com lesões periodontais foram Tannerella forsythia (78,6%), Treponema denticola (78,6%), Fusobacterium nucleatum (64,3%) e Porphyromonas gingivalis (50%). Entretanto, nos 14 ovinos sem lesões periodontais, F. nucleatum (42,8%) foi a bactéria mais detectada. Associação estatisticamente diferente foi observada para Campylobacter rectus, Enterococcus faecium, Prevotella nigrescens, T. forsythia e T. denticola (p<0,05) nos ovinos com periodontite em comparação entre os dois grupos. Aggregatibacter actinomycetemcomitans, Enterococcus faecalis e Porphyromonas gulae não foram detectados em nenhuma das amostras pesquisadas. Conclui-se que C. rectus, E. faecium, P. nigrescens, T. forsythia e T. denticola estão associados às lesões resultantes da periodontite ovina com manifestação clínica grave e F. nucleatum o micro-organismo mais prevalente no biofilme subgengival de animais periodontalmente sadios.(AU)

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Background: Periodontal disease is considered as a diabetes complication and has been suggested that periodontal treatment plus antibiotics should reduce glycated hemoglobin A, by reducing local production of pro inflammatory substances. Objective: To evaluate diabetic patients with periodontal disease under periodontal treatment plus topical antibiotics and reduction of HbA1c, compared to diabetic patients under periodontal treatment without antibiotics. Materials and Methods: Using PUBMED, SCOPUS, WEB OF SCIENCE, EMBASE and Google Scholar data bases, were screened documents from 2008 to 2018. The documents included were the clinical studies, which included non-surgical periodontal treatment plus topical antibiotics, whose outcomes included the HbA1c report. Two independent researchers evaluate title; abstract and bias risk with Downs Black scale and Cochrane tool. Documents with a score higher than 15 on average by the two evaluators were included. Results: Five articles, which find inclusion criteria, were identified. Two documents failed to demonstrate statistically significant effect when compared to non-surgical periodontal therapy alone. Conclusion: In general a modest reduction of HbA1c was identified when using antibiotic therapy.

Antecedentes: La enfermedad periodontal es considerada una complicación del paciente diabético y se ha sugerido que su tratamiento con o sin antibióticos podría aportar a la reducción de la hemoglobina glucosilada, al reducir la producción local de sustancias pro-inflamatorias. Objetivo: Evaluar pacientes diabéticos con enfermedad periodontal que recibieron terapia periodontal no quirúrgica (TPNQ) más antibióticos tópicos y la reducción de la hemoglobina glucosilada, comparados con pacientes que recibieron TPNQ sin antibióticos tópicos. Materiales y métodos: Se utilizaron bases de datos MEDLINE vía PubMed, WEB OF SCIENCE, SCOPUS, EMBASE y Google Scholar utilizando vocabulario controlado y limitando la búsqueda a los años 2008-2018. Los documentos incluidos fueron los estudios clínicos, que incluían tratamiento periodontal no quirúrgico mas antibióticos tópicos, cuyos desenlaces incluyeran el reporte de HbA1c. Estos fueron analizados por dos evaluadores independientes, para identificación de riesgo de sesgo con la escala Downs Black y herramienta Cochrane. Se incluyeron los documentos con puntuación mayor a 15 en promedio de los dos evaluadores. Resultados: Se identificaron 5 documentos que cumplen con los criterios de inclusión. Dos documentos no mostraron diferencias estadísticamente significativas para el cambio de la HbA1c, en comparación con la terapia periodontal sola. Conclusión: En general se evidencia una discreta reducción de HbA1c con la terapia periodontal usando antibióticos tópicos.

RESUMO

Periodontitis is an inflammatory response in a susceptible host caused by complex microbiota, predominantly composed of Gram-negative anaerobic bacteria. Aiming to characterize the subgingival bacterial microbiota associated with ovine periodontitis, polymerase chain reaction (PCR) was performed in subgingival periodontal pocket samples of 14 sheep with severe periodontitis and in subgingival sulcus biofilm of 14 periodontally healthy sheep in search mainly of Gram-negative and Gram-positive microorganisms considered important periodontopathogens. The most prevalent bacteria in the sheep with periodontal lesions were Tannerella forsythia (78.6%), Treponema denticola (78.6%), Fusobacterium nucleatum (64.3%), and Porphyromonas gingivalis (50%), whereas in the healthy sheep, F. nucleatum (42.8%) was the most often detected bacterium. Statistically significant differences were observed for Campylobacter rectus, Enterococcus faecium, Prevotella nigrescens, T. forsythia, and T. denticola (p<0.05) in the sheep with periodontitis in the comparison between groups. Aggregatibacter actinomycetemcomitans, Enterococcus faecalis, and Porphyromonas gulae were not detected in any of the samples analyzed. In conclusion, C. rectus, E. faecium, P. nigrescens, T. forsythia, and T. denticola were associated with severe lesions caused by ovine periodontitis, and F. nucleatum was the most prevalent microorganism in the subgengival sulcus biofilm of healthy sheep.(AU)

Periodontite é a resposta inflamatória de um hospedeiro suscetível causada por complexa microbiota, composta predominantemente por bactérias anaeróbias Gram-negativas. Com o objetivo de caracterizar a microbiota bacteriana subgengival associada à periodontite ovina foi realizada a reação em cadeia da polimerase (PCR) de amostras de biofilme subgengival de 14 ovinos com a enfermidade e 14 ovinos periodontalmente saudáveis, com destaque para micro-organismos Gram-negativos e Gram-positivos considerados importantes periodontopatógenos. As bactérias mais prevalentes em 14 animais com lesões periodontais foram Tannerella forsythia (78,6%), Treponema denticola (78,6%), Fusobacterium nucleatum (64,3%) e Porphyromonas gingivalis (50%). Entretanto, nos 14 ovinos sem lesões periodontais, F. nucleatum (42,8%) foi a bactéria mais detectada. Associação estatisticamente diferente foi observada para Campylobacter rectus, Enterococcus faecium, Prevotella nigrescens, T. forsythia e T. denticola (p<0,05) nos ovinos com periodontite em comparação entre os dois grupos. Aggregatibacter actinomycetemcomitans, Enterococcus faecalis e Porphyromonas gulae não foram detectados em nenhuma das amostras pesquisadas. Conclui-se que C. rectus, E. faecium, P. nigrescens, T. forsythia e T. denticola estão associados às lesões resultantes da periodontite ovina com manifestação clínica grave e F. nucleatum o micro-organismo mais prevalente no biofilme subgengival de animais periodontalmente sadios.(AU)

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ABSTRACT: Periodontitis is an inflammatory response in a susceptible host caused by complex microbiota, predominantly composed of Gram-negative anaerobic bacteria. Aiming to characterize the subgingival bacterial microbiota associated with ovine periodontitis, polymerase chain reaction (PCR) was performed in subgingival periodontal pocket samples of 14 sheep with severe periodontitis and in subgingival sulcus biofilm of 14 periodontally healthy sheep in search mainly of Gram-negative and Gram-positive microorganisms considered important periodontopathogens. The most prevalent bacteria in the sheep with periodontal lesions were Tannerella forsythia (78.6%), Treponema denticola (78.6%), Fusobacterium nucleatum (64.3%), and Porphyromonas gingivalis (50%), whereas in the healthy sheep, F. nucleatum (42.8%) was the most often detected bacterium. Statistically significant differences were observed for Campylobacter rectus, Enterococcus faecium, Prevotella nigrescens, T. forsythia, and T. denticola (p 0.05) in the sheep with periodontitis in the comparison between groups. Aggregatibacter actinomycetemcomitans, Enterococcus faecalis, and Porphyromonas gulae were not detected in any of the samples analyzed. In conclusion, C. rectus, E. faecium, P. nigrescens, T. forsythia, and T. denticola were associated with severe lesions caused by ovine periodontitis, and F. nucleatum was the most prevalent microorganism in the subgengival sulcus biofilm of healthy sheep.

RESUMO: Periodontite é a resposta inflamatória de um hospedeiro suscetível causada por complexa microbiota, composta predominantemente por bactérias anaeróbias Gram-negativas. Com o objetivo de caracterizar a microbiota bacteriana subgengival associada à periodontite ovina foi realizada a reação em cadeia da polimerase (PCR) de amostras de biofilme subgengival de 14 ovinos com a enfermidade e 14 ovinos periodontalmente saudáveis, com destaque para micro-organismos Gram-negativos e Gram-positivos considerados importantes periodontopatógenos. As bactérias mais prevalentes em 14 animais com lesões periodontais foram Tannerella forsythia (78,6%), Treponema denticola (78,6%), Fusobacterium nucleatum (64,3%) e Porphyromonas gingivalis (50%). Entretanto, nos 14 ovinos sem lesões periodontais, F. nucleatum (42,8%) foi a bactéria mais detectada. Associação estatisticamente diferente foi observada para Campylobacter rectus, Enterococcus faecium, Prevotella nigrescens, T. forsythia e T. denticola (p 0,05) nos ovinos com periodontite em comparação entre os dois grupos. Aggregatibacter actinomycetemcomitans, Enterococcus faecalis e Porphyromonas gulae não foram detectados em nenhuma das amostras pesquisadas. Conclui-se que C. rectus, E. faecium, P. nigrescens, T. forsythia e T. denticola estão associados às lesões resultantes da periodontite ovina com manifestação clínica grave e F. nucleatum o micro-organismo mais prevalente no biofilme subgengival de animais periodontalmente sadios.