Effect of adjuvant probiotic therapy (Lactobacillus reuteri) in the treatment of periodontitis associated with diabetes mellitus: clinical, controlled, and randomized study.

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.

Evaluation of the Microbial Profile on the Polydioxanone Membrane and the Collagen Membrane Exposed to Multi-Species Subgingival Biofilm: An In Vitro Study.

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.

Levels of HIV-1 in subgingival biofilm of HIV-infected patients.

AIM: The aims of the current study were to compare the levels of HIV-1 in the subgingival biofilm (SHVL) between detectable and undetectable plasmatic HIV-1 viral load (PHVL) in HIV-infected patients as well as to determine the association of SHVL with PHVL and clinical periodontal parameters. MATERIAL AND METHODS: Forty-one HIV-infected individuals were divided into two groups: detectable (21) and undetectable (20) PHVL. Subgingival biofilm samples were obtained for detection and quantification of HIV-1 by real-time RT-PCR. To estimate the effect of co-variables on the outcome undetectable SHVL, the Generalized Estimation Equation (GEE) was employed. RESULTS: Detectable SHVL was observed only in the detectable PHVL group and the detection of the HIV-1 was observed in 40% of these individuals. In the bivariate analysis between co-variables from the individual level and the outcome SHVL, significant difference was observed only for the CD4+ T lymphocytes levels (p = 0.017). The multiple logistic model demonstrated that only CD4+ T lymphocytes levels had a significant effect on the outcome undetectable SHVL [OR 8.85 (CI 3.6-9.2), p = 0.002]. CONCLUSION: HIV-1 can be detected and quantified in the subgingival biofilm of HIV-infected individuals, but these findings are not associated with PHVL and periodontal clinical parameters.

Prevalence of Pseudomonas aeruginosa and Acinetobacter spp. in subgingival biofilm and saliva of subjects with chronic periodontal infection.

P. aeruginosa and Acinetobacter spp. are important pathogens associated with late nosocomial pneumonia in hospitalized and institutionalized individuals. The oral cavity may be a major source of these respiratory pathogens, particularly in the presence of poor oral hygiene and periodontal infection. This study investigated the prevalence of P. aeruginosa and Acinetobacter spp. in subgingival biofilm and saliva of subjects with periodontal disease or health. Samples were obtained from 55 periodontally healthy (PH) and 169 chronic periodontitis (CP) patients. DNA was obtained from the samples and detection of P. aeruginosa and Acinetobacter spp. was carried out by multiplex and nested PCR. P. aeruginosa and Acinetobacter spp. were detected in 40% and 45% of all samples, respectively. No significant differences in the distribution of these microorganisms between men and women, subgingival biofilm and saliva samples, patients ≤ 35 and > 35 years of age, and smokers and non-smokers were observed regardless periodontal status (p > 0.05). In contrast, the frequencies of P. aeruginosa and Acinetobacter spp. in saliva and biofilm samples were significantly greater in CP than PH patients (p < 0.01). Smokers presenting P. aeruginosa and high frequencies of supragingival plaque were more likely to present CP than PH. P. aeruginosa and Acinetobacter spp. are frequently detected in the oral microbiota of CP. Poor oral hygiene, smoking and the presence of P. aeruginosa are strongly associated with periodontitis.

Prevalence of Pseudomonas aeruginosa and Acinetobacter spp. in subgingival biofilm and saliva of subjects with chronic periodontal infection

P. aeruginosa and Acinetobacter spp. are important pathogens associated with late nosocomial pneumonia in hospitalized and institutionalized individuals. The oral cavity may be a major source of these respiratory pathogens, particularly in the presence of poor oral hygiene and periodontal infection. This study investigated the prevalence of P. aeruginosa and Acinetobacter spp. in subgingival biofilm and saliva of subjects with periodontal disease or health. Samples were obtained from 55 periodontally healthy (PH) and 169 chronic periodontitis (CP) patients. DNA was obtained from the samples and detection of P. aeruginosa and Acinetobacter spp. was carried out by multiplex and nested PCR. P. aeruginosa and Acinetobacter spp. were detected in 40% and 45% of all samples, respectively. No significant differences in the distribution of these microorganisms between men and women, subgingival biofilm and saliva samples, patients < 35 and > 35 years of age, and smokers and non-smokers were observed regardless periodontal status (p > 0.05). In contrast, the frequencies of P. aeruginosa and Acinetobacter spp. in saliva and biofilm samples were significantly greater in CP than PH patients (p < 0.01). Smokers presenting P. aeruginosa and high frequencies of supragingival plaque were more likely to present CP than PH. P. aeruginosa and Acinetobacter spp. are frequently detected in the oral microbiota of CP. Poor oral hygiene, smoking and the presence of P. aeruginosa are strongly associated with periodontitis.

Prevalence of Pseudomonas aeruginosa and Acinetobacter spp. in subgingival biofilm and saliva of subjects with chronic periodontal infection

P. aeruginosa and Acinetobacter spp. are important pathogens associated with late nosocomial pneumonia in hospitalized and institutionalized individuals. The oral cavity may be a major source of these respiratory pathogens, particularly in the presence of poor oral hygiene and periodontal infection. This study investigated the prevalence of P. aeruginosa and Acinetobacter spp. in subgingival biofilm and saliva of subjects with periodontal disease or health. Samples were obtained from 55 periodontally healthy (PH) and 169 chronic periodontitis (CP) patients. DNA was obtained from the samples and detection of P. aeruginosa and Acinetobacter spp. was carried out by multiplex and nested PCR. P. aeruginosa and Acinetobacter spp. were detected in 40% and 45% of all samples, respectively. No significant differences in the distribution of these microorganisms between men and women, subgingival biofilm and saliva samples, patients < 35 and > 35 years of age, and smokers and non-smokers were observed regardless periodontal status (p > 0.05). In contrast, the frequencies of P. aeruginosa and Acinetobacter spp. in saliva and biofilm samples were significantly greater in CP than PH patients (p < 0.01). Smokers presenting P. aeruginosa and high frequencies of supragingival plaque were more likely to present CP than PH. P. aeruginosa and Acinetobacter spp. are frequently detected in the oral microbiota of CP. Poor oral hygiene, smoking and the presence of P. aeruginosa are strongly associated with periodontitis.

Associação entre a carga viral plasmática e a carga viral subgengival no biofilme em pacientes infectados pelo HIV-1 / Association between plasmatic viral load and subgingival viral load in biofilm in HIV-1 infected patients

Objetivo: Investigar a associação entre carga viral no biofilme subgengival (CVS) e carga viral plasmática (CVP) do HIV-1 e os parâmetros clínicos periodontais em indivíduos infectados pelo HIV. Metodologia: Quarenta e um pacientes com sorologia positiva para HIV-1 foram distribuídos em dois grupos: 20 com CVP detectável e 21 com CVP indetectável (< 50 cópias/mL). Amostras do biofilme subgengival foram coletadas de 12 sítios de cada paciente com periodontite, sendo 6 sítios com as maiores PBS, preferencialmente não adjacentes, e 6 sítios com periodonto sadio. Nos pacientes com saúde periodontal, foram coletadas 6 amostras de sítios periodontais sadios aleatórios. Das 6 amostras dos sítios com maiores PBS dos pacientes com periodontite, foram realizados 3 pools, com 2 amostras cada, de sítios com parâmetros clínicos semelhantes. O mesmo foi feito para as 6 amostras de periodontos sadios (tanto para os pacientes com periodontite como para aqueles com saúde periodontal), resultando em 3 pools. A detecção e quantificação da CVS foram realizadas através do PCR em Tempo Real. Os níveis de linfócitos T CD4+, T CD8+, neutrófilos e CVP foram obtidos dos resultados desses exames presentes nos prontuários médicos de cada paciente. Diferenças significativas entre os grupos para todos os parâmetros estudados foram avaliadas pelos testes Qui-quadrado, exato de Fisher e Mann-Whitney. O modelo de regressão utilizado foi a Equação de Estimação Generalizada (GEE) para testar a associação entre as co-variáveis e a CVS. Resultados: A maioria dos indivíduos era composta por homens (51,2%), não usuários de drogas (85,4%), não tabagistas (63,4%) e não consumidores diários de álcool (87,8%). Os dados laboratoriais demonstraram diferenças significantes entre os dois grupos estudados somente para os níveis de linfócitos T CD4+ (p = 0,038), porém não houve diferença estatisticamente significante entre os grupos em relação aos parâmetros clínicos periodontais. Não houve associação estatisticamente significante entre CV plasmática e CV subgengival, sendo a contagem de linfócitos TCD4+ a única co-variável que apresentou efeito estatisticamente significante sobre o desfecho CV subgengival indetectável (p = 0,017). A interpretação dos resultados demonstrou que indivíduos com níveis mais altos de linfócitos T CD4+ (> 500 células/mm3 ) tem uma chance muito maior de apresentar CV subgengival indetectável em relação aos indivíduos com níveis de linfócito T CD4+ < 200 células/mm3 . Conclusão: Não há associação entre a carga viral no biofilme subgengival e a carga viral plasmática do HIV-1 e os parâmetros clínicos periodontais em pacientes infectados pelo HIV.(AU)

Aim: To investigate the association between subgingival viral load in biofilm and plasmatic viral load and clinical periodontal parameters in HIV-1 infected patients. Methods: Fortyone HIV-positive patients were divided into two groups: detectable and undetectable plasmatic viral load (< 50 copies/mL). Subgingival biofilm samples were collected from 12 sites in each patient with periodontitis, 6 sites with the deepest PBS, preferably not adjacent, and 6 sites with healthy periodontium. In patients with periodontal health, 6 samples were collected from random healthy periodontal sites. Of the six samples of periodontal lesions were performed 3 pools, with 2 samples from each site with similar clinical parameters. The same was done with the 6 samples from healthy periodontium, resulting in three pools. Detection and quantification of subgingival viral load were determined by Real Time PCR. The levels of CD4, CD8, neutrophils and plasmatic viral load were obtained from the medical records of each patient. Significant differences between groups for all parameters were evaluated by Chi-square, Fisher's exact test and Mann-Whitney. The utilized regression model consisted of a test called Generalized Estimation Equation (GEE) to test the association between the co-variables and the subgingival viral load. Results: The majority of the patients were male (51,2%), non-drug users (85,4%), non-smokers (63,4%) and nonalcohol users (87,8%). The laboratory data demonstrated significant differences between groups only for levels of TCD4+ cells (p=0,038), however there was no significant difference between groups for clinical periodontal parameters. There was no statistically significant association between plasmatic viral load and subgingival viral load, and only the levels of TCD4+ cells demonstrated significant effect on the undetectable subgingival viral load (p=0,017). The interpretation of the results demonstrated that patients with higher levels of TCD4+ cells (> 500 cells/mm3 ) have a much higher chance of presenting undetectable subgingival viral load compared to patients with levels of TCD4+ cells < 200 cells/mm3. Conclusion: There is no association between subgingival viral load in biofilm and plasmatic viral load and clinical periodontal parameters in HIV-1 infected patients(AU)

Genetic relatedness of subgingival and buccal Candida dubliniensis isolates in immunocompetent subjects assessed by RAPD-PCR.

BACKGROUND: It is recognized that Candida dubliniensis commonly colonizes oral and subgingival sites in immunocompetent subjects with periodontal disease. OBJECTIVE: Since there are few data available on genetic characterization of C. dubliniensis in periodontal pockets and other oral sites, the aim of this study was to characterize subgingival and mucosal C. dubliniensis isolates recovered from immunocompetent subjects and to assay the genetic similarity of such isolates from both niches in the same patient by random amplified polymorphic DNA (RAPD). DESIGN: C. dubliniensis recovered from subgingival plaque and from buccal cavity samples were studied in 240 immunocompetent non-smoking individuals. Arbitrary amplification was carried out by RAPD-polymerase chain reaction (PCR). RESULTS: RAPD analysis showed identical genotypes of C. dubliniensis in different sampling sites (buccal cavity and subgingival areas) in eight of 10 patients except for those derived from two participants who presented presumably unrelated isolates. CONCLUSIONS: On the basis of the findings presented, the origin of the colonization of C. dubliniensis in subgingival biofilm seems to be the buccal cavity in a single patient. Consequently, it may be assumed that most of C. dubliniensis in these sites arise from the endogenous commensal strains.

Prevalence of leukotoxic genotypes of Actinobacillus actinomycetemcomitans in Brazilians with chronic periodontitis

Actinobacillus actinomycetemcomitans is considered a major etiologic agent of aggressive periodontitis but this species has also been associated with other forms of periodontal disease. Further, highly leukotoxic strains are related to severity of disease. This investigation determined the prevalence of A. actinomycetemcomitans and the occurrence of the leukotoxin gene 530-bp deletion in Brazilian subjects with chronic periodontitis. Twenty periodontally healthy and 20 chronic periodontitis subjects were selected. Full-mouth clinical examination was carried out at 6 sites/tooth. Subgingival biofilm samples were collected from the 3 deepest sites and 3 healthy sites from periodontitis subjects, as well as from 3 sites of individuals with periodontal health. A. actinomycetemcomitans and the genetic deletion were determined by the polymerase chain reaction. Significant differences were sought by Mann-Whitney, Chi-square, and Wilcoxon sign tests. Periodontitis subjects presented a higher prevalence (75%) of A. actinomycetemcomitans than individuals with health (45%) (p = 0.053). A mean frequency of 57.5% of A. actinomycetemcomitans - positive sites was observed in the periodontitis group. Of those, 75% were diseased, whereas 40% were healthy sites (p = 0.0001). Healthy subjects showed a mean frequency of 35% of positive sites. In contrast, the genetic deletion was detected only in 4 diseased sites from 2 chronic periodontitis patients. A high prevalence of A. actinomycetemcomitans was observed in Brazilians with chronic periodontitis. However, the leukotoxin gene 530-bp deletion was rarely detected in the subgingival biofilm of these subjects.

Actinobacillus actinomycetemcomitans tem sido associado com diferentes formas de doenças periodontais, mas tal espécie é considerada o principal agente etiológico da doença periodontal agressiva. Algumas cepas de A. actinomycetemcomitans apresentam uma deleção de 530 pb na região promotora do operon do gene da leucotoxina, produzindo assim maiores quantidades desta toxina. Tal fato pode ter um importante papel na patogênese das doenças periodontais. A proposta do presente estudo foi determinar a prevalência do A. actinomycetemcomitans em amostras de biofilme subgengival de indivíduos brasileiros com periodontite crônica e saúde periodontal; bem como avaliar a distribuição do tipo genético leucotóxico desta espécie nestes indivíduos. Vinte pacientes com periodontite crônica e 20 controles com saúde periodontal foram selecionados. O exame clínico periodontal foi realizado em 6 sítios/dente em todos os dentes. Amostras de biofilme subgengival foram coletadas de 3 sítios com a maior profundidade de bolsa e 3 sítios sem doença dos pacientes com periodontite, assim como de 3 sítios aleatórios dos controles. A detecção do A. actinomycetemcomitans e a ocorrência da deleção genética foram realizadas utilizando a técnica de PCR diretamente nas amostras de biofilme. Pacientes com periodontite crônica apresentaram uma alta prevalência de A. actinomycetemcomitans (75%) quando comparados aos indivíduos sem doença periodontal (45%), porém essa diferença não foi significativa (p = 0.053). Foi observada uma freqüência média de 57.5% de sítios com A. actinomycetemcomitans no grupo com periodontite. Destes sítios, 75% eram sítios com doença, enquanto 40% eram sítios saudáveis (p = 0.0001). Indivíduos sem doença periodontal apresentaram uma freqüência média de 35% de sítios com A. actinomycetemcomitans. A deleção de 530-pb foi encontrada somente em 4 sítios doentes de 2 pacientes com periodontite crônica. Uma alta prevalência de A. actinomycetemcomitans foi observada em pacientes Brasileiros com doença periodontal crônica. Entretanto, a deleção de 530 pb no gene da leucotoxina foi raramente detectada no biofilme subgengival desses indivíduos.