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OBJECTIVE: The aim of this study was to optimize the sensitivity, specificity and cost-effectiveness of the RNA-Oligonucleotide Quantification Technique (ROQT) in order to identify periodontal pathogens that remain unrecognized or uncultured in the oral microbiome. DESIGN: Total nucleic acids (TNA) were extracted from subgingival biofilm samples using an automated process. RNA, DNA and Locked Nucleic Acid (LNA) digoxigenin-labeled oligonucleotide probes targeting 5 cultivated/named species and 16 uncultivated or unnamed bacterial taxa were synthesized. Probe specificity was determined by targeting 96 oral bacterial species; sensitivity was assessed using serial dilutions of reference bacterial strains. Different stringency temperatures were compared and new standards were tested. The tested conditions were evaluated analyzing samples from periodontally healthy individuals, and patients with moderate or severe periodontitis. RESULTS: The automated extraction method at 63°C along with LNA-oligunucleotides probes, and use of reverse RNA sequences for standards yielded stronger signals without cross-reactions. In the pilot clinical study, the most commonly detected uncultivated/unrecognized species were Selenomonas sp. HMT 134, Prevotella sp. HMT 306, Desulfobulbus sp. HMT 041, Synergistetes sp. HMT 360 and Bacteroidetes HMT 274. In the cultivated segment of the microbiota, the most abundant taxa were T. forsythia HMT 613 and Fretibacterium fastidiosum (formerly Synergistetes) HMT 363. CONCLUSIONS: In general, samples from severe patients had the greatest levels of organisms. Classic (T. forsythia, P. gingivalis) and newly proposed (F. alocis and Desulfobulbus sp. HMT 041) pathogens were present in greater amounts in samples from severe periodontitis sites, followed by moderate periodontitis sites.
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Placa Dentária , Periodontite , Humanos , Placa Dentária/microbiologia , RNA , Periodontite/microbiologia , Oligonucleotídeos , DNA Bacteriano , Porphyromonas gingivalis/genéticaRESUMO
BACKGROUND: Oral microbiome dysbiosis is linked to overt inflammation of tooth-supporting tissues, leading to periodontitis, an oral condition that can cause tooth and bone loss. Microbiome dysbiosis has been described as a disruption in the symbiotic microbiota composition's stability that could adversely affect the host's health status. However, the precise microbiome dynamics that lead to dysbiosis and the progression of the disease are largely unknown. The objective of our study was to investigate the long-term dynamics of periodontitis progression and its connection to dysbiosis. RESULTS: We studied three different teeth groups: sites that showed disease progression, sites that remained stable during the study, and sites that exhibited a cyclic deepening followed by spontaneous recovery. Time-series analysis revealed that communities followed a characteristic succession of bacteria clusters. Stable and fluctuating sites showed high asynchrony in the communities (i.e., different species responding dissimilarly through time) and a reordering of the communities where directional changes dominated (i.e., sample distance increases over time) in the stable sites but not in the fluctuating sites. Progressing sites exhibited low asynchrony and convergence (i.e., samples distance decreases over time). Moreover, new species were more likely to be recruited in stable samples if a close relative was not recruited previously. In contrast, progressing and fluctuating sites followed a neutral recruitment model, indicating that competition between closely related species is a significant component of species-species interactions in stable samples. Finally, periodontal treatment did not select similar communities but stabilized α-diversity, centered the abundance of different clusters to the mean, and increased community rearrangement. CONCLUSIONS: Here, we show that ecological principles can define dysbiosis and explain the evolution and outcomes of specific microbial communities of the oral microbiome in periodontitis progression. All sites showed an ecological succession in community composition. Stable sites were characterized by high asynchrony, a reordering of the communities where directional changes dominated, and new species were more likely to be recruited if a close relative was not recruited previously. Progressing sites were characterized by low asynchrony, community convergence, and a neutral model of recruitment. Finally, fluctuating sites were characterized by high asynchrony, community convergence, and a neutral recruitment model.
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Microbiota , Periodontite , Bactérias , Progressão da Doença , Disbiose , HumanosRESUMO
The case of Phineas Gage is an integral part of medical folklore. His accident still causes astonishment and curiosity and can be considered as the case that most influenced and contributed to the nineteenth century's neuropsychiatric discussion on the mind-brain relationship and brain topography. It was perhaps the first case to suggest the role of brain areas in determining personality and which specific parts of the brain, when affected, can induce specific mental changes. In addition, his case contributed to the emergence of the scientific approaches that would later culminate in psychosurgery. Gage is a fixed element in the studies of neurology, psychology, and neuroscience, having been solidified as one of the greatest medical curiosities of all time, deserving its prominence.
O caso de Phineas Gage é parte integrante do folclore médico. Seu acidente ainda causa espanto e curiosidade, e pode ser considerado como o caso que mais influenciou e contribuiu para a discussão neuropsiquiátrica do século XIX sobre a relação mente-cérebro e topografia cerebral. Foi talvez o primeiro caso a sugerir o papel de áreas cerebrais na determinação da personalidade e que partes específicas do cérebro, quando afetadas, podem induzir mudanças mentais específicas. Além disso, seu caso contribuiu para o surgimento de abordagens cientificas que culminariam posteriormente na psicocirurgia. Gage é um elemento fixo nos estudos de neurologia, psicologia e neurociências, tendo sido solidificado como uma das grandes curiosidades médicas de todos os tempos que merece seu destaque.
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Hermann Rorschach was a Swiss psychiatrist and psychoanalyst, best known for developing a projective test known as the Rorschach inkblot test, a test designed to reflect unconscious parts of the personality that project into the visual stimuli generated by the inkblots, allowing a psychodiagnosis to be established. The technique he developed has been applied since 1921 in a number of countries. Although it has long remained controversial and divided opinions, this did not prevent it from overcoming the barriers of science to have a major influence on pop culture, resulting in an undeniable legacy for the development of Psychiatry in the nineteenth century.
Hermann Rorschach foi um psiquiatra e psicanalista suíço, mais conhecido por desenvolver um teste projetivo conhecido como o teste da mancha de tinta de Rorschach, um teste desenhado para refletir partes inconscientes da personalidade que se projetam nos estímulos visuais gerado pelas manchas de tinta, o que possibilitaria estabelecer um psicodiagnóstico. A técnica que ele desenvolveu tem sido aplicada desde 1921 em vários países, e embora por muito tenha permanecido polêmica e dividido opiniões, isso não a impediu de ultrapassar as barreiras da ciência para alcançando massivamente a cultura pop, resultando em um legado inegável para o desenvolvimento da Psiquiatria no século XIX.
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OBJECTIVE: The aim of this study was to evaluate the levels of Interleukin-1α (IL-1α), Interleukin-1ß (IL-1ß), Interleukin-1 receptor antagonist (IL-1Ra), Interleukin-10 (IL-10), Interleukin-13 (IL-13), Vascular endothelial growth factor (VEGF), Granulocyte-colony stimulating factor (G-CSF), and Growth related oncogene (GRO) in the peri-miniscrew implant crevicular fluid (MICF) under orthodontic loading. DESIGN: The study sample comprised 14 miniscrews immediately loaded and 17 unloaded ones. A load of 200gF was immediately applied to the miniscrews in the loaded group after the placement surgery. Peri-miniscrew implant crevicular fluid was collected at baseline, at day 7, and at day 21. The levels of the biomarkers were measured using a multiplexed bead immunoassay. Intergroup comparisons were made using Mann-Whitney test. Friedman and Dunn's multiple comparison tests were used to evaluate intragroup differences over time. RESULTS: Although no statistical differences were observed between the groups at any time point for any of the 8 biomarkers evaluated, there was a statistically significant increase (p < 0.02) in the levels of all the biomarkers over time on both groups. CONCLUSIONS: An immediate loading of 200gF does not alter the balance in the inflammatory response in peri-miniscrew tissues.
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Parafusos Ósseos , Interleucinas/metabolismo , Procedimentos de Ancoragem Ortodôntica , Adolescente , Adulto , Biomarcadores/metabolismo , Quimiocina CXCL1/metabolismo , Feminino , Fator Estimulador de Colônias de Granulócitos/metabolismo , Humanos , Inflamação , Masculino , Técnicas de Movimentação Dentária , Fator A de Crescimento do Endotélio Vascular/metabolismo , Adulto JovemRESUMO
A new periodontitis classification scheme has been adopted, in which forms of the disease previously recognized as "chronic" or "aggressive" are now grouped under a single category ("periodontitis") and are further characterized based on a multi-dimensional staging and grading system. Staging is largely dependent upon the severity of disease at presentation as well as on the complexity of disease management, while grading provides supplemental information about biological features of the disease including a history-based analysis of the rate of periodontitis progression; assessment of the risk for further progression; analysis of possible poor outcomes of treatment; and assessment of the risk that the disease or its treatment may negatively affect the general health of the patient. Necrotizing periodontal diseases, whose characteristic clinical phenotype includes typical features (papilla necrosis, bleeding, and pain) and are associated with host immune response impairments, remain a distinct periodontitis category. Endodontic-periodontal lesions, defined by a pathological communication between the pulpal and periodontal tissues at a given tooth, occur in either an acute or a chronic form, and are classified according to signs and symptoms that have direct impact on their prognosis and treatment. Periodontal abscesses are defined as acute lesions characterized by localized accumulation of pus within the gingival wall of the periodontal pocket/sulcus, rapid tissue destruction and are associated with risk for systemic dissemination.
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Doenças Periodontais , Periodontite , Consenso , Humanos , Bolsa Periodontal , PeriodontoRESUMO
A new periodontitis classification scheme has been adopted, in which forms of the disease previously recognized as "chronic" or "aggressive" are now grouped under a single category ("periodontitis") and are further characterized based on a multi-dimensional staging and grading system. Staging is largely dependent upon the severity of disease at presentation as well as on the complexity of disease management, while grading provides supplemental information about biological features of the disease including a history-based analysis of the rate of periodontitis progression; assessment of the risk for further progression; analysis of possible poor outcomes of treatment; and assessment of the risk that the disease or its treatment may negatively affect the general health of the patient. Necrotizing periodontal diseases, whose characteristic clinical phenotype includes typical features (papilla necrosis, bleeding, and pain) and are associated with host immune response impairments, remain a distinct periodontitis category. Endodontic-periodontal lesions, defined by a pathological communication between the pulpal and periodontal tissues at a given tooth, occur in either an acute or a chronic form, and are classified according to signs and symptoms that have direct impact on their prognosis and treatment. Periodontal abscesses are defined as acute lesions characterized by localized accumulation of pus within the gingival wall of the periodontal pocket/sulcus, rapid tissue destruction and are associated with risk for systemic dissemination.
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Peri-Implantite , Doenças Periodontais , Periodontite , Consenso , Humanos , PeriodontoRESUMO
Antibodies to leukotoxin A are markers that link Aggregatibacter actinomycetemcomitans-associated periodontitis and rheumatoid arthritis.
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Artrite Reumatoide , Periodontite , Aggregatibacter actinomycetemcomitans , Autoimunidade , HumanosRESUMO
AIM: The goal of the present longitudinal cohort study was to examine patterns of periodontal disease progression at progressing sites and subjects defined based on linear mixed models (LMM) of clinical attachment loss (CAL). MATERIALS AND METHODS: A total of 113 periodontally healthy and 302 periodontitis subjects had their CAL calculated bimonthly for 12 months. LMMs were fitted for each site and the predicted CAL levels used to categorize their progression state. Participants were grouped based on the number of progressing sites into unchanged, transitional and active subjects. Patterns of periodontal disease progression were explored using descriptive statistics. RESULTS: Progression occurred primarily at molars (50% of progressing sites) and inter-proximal sites (72%), affected a higher proportion of deep than shallow sites (2.7% versus 0.7%), and pocketing was the main mode of progression (49%). We found a low level of agreement (47%) between the LMM and traditional approaches to determine progression such as change in CAL ≥3 mm. Fourteen per cent of subjects were classified as active and among those 93% had periodontitis. The annual mean rate of progression for the active subjects was 0.35 mm/year. CONCLUSION: Progressing sites and subjects defined based on LMMs presented patterns of disease progression similar to those previously reported in the literature.
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Progressão da Doença , Modelos Lineares , Perda da Inserção Periodontal/complicações , Doenças Periodontais/etiologia , Adulto , Estudos de Coortes , Feminino , Humanos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Estudos ProspectivosRESUMO
AIM: To compare the microbiome of healthy (H) and diseased (P) peri-implant sites and determine the core peri-implant microbiome. MATERIALS AND METHODS: Submucosal biofilms from 32 H and 35 P sites were analysed using 16S rRNA sequencing (MiSeq, Illumina), QIIME and HOMINGS. Differences between groups were determined using principal coordinate analysis (PCoA), t tests and Wilcoxon rank sum test and FDR-adjusted. The peri-implant core microbiome was determined. RESULTS: PCoA showed partitioning between H and P at all taxonomic levels. Bacteroidetes, Spirochetes and Synergistetes were higher in P, while Actinobacteria prevailed in H (p < .05). Porphyromonas and Treponema were more abundant in P while Rothia and Neisseria were higher in H (p < .05). The core peri-implant microbiome contained Fusobacterium, Parvimonas and Campylobacter sp. T. denticola, and P. gingivalis levels were higher in P, as well as F. alocis, F. fastidiosum and T. maltophilum (p < .05). CONCLUSION: The peri-implantitis microbiome is commensal-depleted and pathogen-enriched, harbouring traditional and new pathogens. The core peri-implant microbiome harbours taxa from genera often associated with periodontal inflammation.
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Bactérias/classificação , Bactérias/isolamento & purificação , Implantes Dentários/microbiologia , Microbiota/genética , Peri-Implantite/microbiologia , Adulto , Idoso , Perda do Osso Alveolar/microbiologia , Bactérias/genética , Carga Bacteriana , Sequência de Bases , Biofilmes/crescimento & desenvolvimento , Estudos de Casos e Controles , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Feminino , Humanos , Masculino , Consórcios Microbianos/genética , Pessoa de Meia-Idade , Bolsa Periodontal/microbiologia , RNA Ribossômico 16S/genéticaRESUMO
BACKGROUND: The single nucleotide polymorphism (SNP) context of a previously identified periodontitis-associated locus is investigated, and its association with microbial, biologic, and periodontal disease clinical parameters is examined. METHODS: A 200-kb spanning region of 1q12 previously highlighted in a genome-wide association scan among 4,766 European American individuals (SNP rs1633266) was annotated. Two haplotype blocks were selected. Association of these polymorphisms with data on microbial plaque composition, gingival crevicular fluid (GCF)-interleukin (IL)-1ß levels, and clinical parameters of periodontal disease were examined. Descriptive analysis of IFI16 and AIM2 protein expression in gingival tissues from healthy individuals (n = 2) and individuals with chronic periodontitis (n = 2) was done via immunohistochemistry. RESULTS: The highlighted locus is a 100-kb region containing the interferon γ-inducible protein 16 (IFI16) and absent in melanoma 2 (AIM2) genes. Two haplotype blocks, rs6940 and rs1057028, were significantly associated with increased extent bleeding on probing and levels of microorganisms Porphyromonas gingivalis, Tannerella forsythia, and Campylobacter rectus (P ≤0.05). Haplotype block rs1057028 was also significantly associated with pathogens Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans, increased GCF-IL-1ß levels, and extent of probing depth ≥4 mm (P ≤0.05). Prevalence of severe periodontitis (biofilm-gingival interface P3 classification) was positively associated with haplotype block rs1057028. Similar trends were observed for haplotype block rs1057028. IFI16 and AIM2 protein expression was observed in multiple cell types of gingival tissues, including inflammatory cells. CONCLUSION: This study found IFI16 and AIM2 SNPs associated with higher levels of periodontal microorganisms and an increased percentage of periodontal disease clinical parameters, suggesting the need for functional studies and additional fine-mapping of variants in the 1q12-locus.
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Periodontite Crônica/genética , Proteínas de Ligação a DNA/genética , Gengiva/metabolismo , Proteínas Nucleares/genética , Fosfoproteínas/genética , Polimorfismo de Nucleotídeo Único , Idoso , Aggregatibacter actinomycetemcomitans/isolamento & purificação , Periodontite Crônica/metabolismo , Periodontite Crônica/microbiologia , Proteínas de Ligação a DNA/metabolismo , Placa Dentária/microbiologia , Feminino , Fusobacterium nucleatum/isolamento & purificação , Estudo de Associação Genômica Ampla , Gengiva/microbiologia , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas Nucleares/metabolismo , Fosfoproteínas/metabolismo , População BrancaRESUMO
BACKGROUND: Chronic periodontitis is controlled without antibiotics by scaling and root planing (SRP) to remove dental biofilm. It has been previously reported that the epithelial barrier to bacterial proinflammatory products is impaired when biofilm lysine falls below the minimal content of normal blood plasma. Aims were to examine whether being refractory and requiring antibiotics to supplement SRP were associated with low biofilm lysine contents. METHODS: Sixteen patients with periodontitis and six periodontally healthy volunteers (HVs) (respective mean ages: 57 ± 6 and 36 ± 8 years) were examined. Patients with periodontitis received SRP and surgery, and HVs received prophylaxis. At quarterly maintenance or prophylaxis visits during the subsequent year, therapeutic response was good (GR, n = 9) or poor (PR, n = 7; including five cigarette smokers). Biofilm cadaverine, lysine, and other amino acid (AA) contents were determined by liquid chromatography. Cadaverine mole fraction of lysine plus cadaverine (CF) indicated biofilm lysine decarboxylase activity. RESULTS: Biofilm lysine was 0.19 ± 0.10 and 0.20 ± 0.09 µmol/mg in GRs and HVs, but 0.07 ± 0.03 µmol/mg in PRs (Kruskal-Wallis: P <0.01). All AAs were depleted in biofilm from smokers, but only lysine was depleted in biofilm from non-smokers. CF was inversely associated with clinical attachment level (CAL) at baseline before therapy in all patients (R2 = 0.28, P <0.01) and with CAL change after therapy in GR (R2 = 0.49, P <0.05). Lysine and cadaverine contents discriminated PRs from GRs and HVs (Wilks' λ = 0.499, P <0.012). CONCLUSIONS: Refractory responses requiring antibiotic therapy result from smoking and/or microbial infections that starve the biofilm and epithelial attachment of lysine. Biofilm CF is associated with periodontitis severity pretherapy and extent of therapeutic response post-therapy.
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Biofilmes , Periodontite Crônica/terapia , Lisina/análise , Adulto , Antibacterianos/uso terapêutico , Cadaverina/análise , Cromatografia Líquida , Periodontite Crônica/microbiologia , Terapia Combinada , Raspagem Dentária , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Aplainamento Radicular , Fumar/efeitos adversosRESUMO
A bacterial etiology of rheumatoid arthritis (RA) has been suspected since the beginnings of modern germ theory. Recent studies implicate mucosal surfaces as sites of disease initiation. The common occurrence of periodontal dysbiosis in RA suggests that oral pathogens may trigger the production of disease-specific autoantibodies and arthritis in susceptible individuals. We used mass spectrometry to define the microbial composition and antigenic repertoire of gingival crevicular fluid in patients with periodontal disease and healthy controls. Periodontitis was characterized by the presence of citrullinated autoantigens that are primary immune targets in RA. The citrullinome in periodontitis mirrored patterns of hypercitrullination observed in the rheumatoid joint, implicating this mucosal site in RA pathogenesis. Proteomic signatures of several microbial species were detected in hypercitrullinated periodontitis samples. Among these, Aggregatibacter actinomycetemcomitans (Aa), but not other candidate pathogens, induced hypercitrullination in host neutrophils. We identified the pore-forming toxin leukotoxin A (LtxA) as the molecular mechanism by which Aa triggers dysregulated activation of citrullinating enzymes in neutrophils, mimicking membranolytic pathways that sustain autoantigen citrullination in the RA joint. Moreover, LtxA induced changes in neutrophil morphology mimicking extracellular trap formation, thereby releasing the hypercitrullinated cargo. Exposure to leukotoxic Aa strains was confirmed in patients with RA and was associated with both anticitrullinated protein antibodies and rheumatoid factor. The effect of human lymphocyte antigen-DRB1 shared epitope alleles on autoantibody positivity was limited to RA patients who were exposed to Aa These studies identify the periodontal pathogen Aa as a candidate bacterial trigger of autoimmunity in RA.
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Aggregatibacter actinomycetemcomitans , Anticorpos Antiproteína Citrulinada/imunologia , Artrite Reumatoide/imunologia , Citrulina/química , Infecções por Pasteurellaceae/imunologia , Periodontite/microbiologia , Adulto , Artrite Reumatoide/microbiologia , Autoantígenos/química , Estudos de Casos e Controles , Doença Crônica , Ensaios Clínicos como Assunto , Feminino , Cadeias HLA-DRB1/genética , Humanos , Masculino , Pessoa de Meia-Idade , Neutrófilos/imunologia , Periodontite/imunologia , Estudos ProspectivosRESUMO
Periodontitis is a biofilm-induced inflammatory disease characterized by dysbiosis of the commensal periodontal microbiota. It is unclear how natural regulation of inflammation affects the periodontal biofilm. Promoters of active resolution of inflammation, including resolvin E1 (RvE1), effectively treat inflammatory periodontitis in animal models. The goals of this study were 1) to compare periodontal tissue gene expression in different clinical conditions, 2) to determine the impact of local inflammation on the composition of subgingival bacteria, and 3) to understand how inflammation impacts these changes. Two clinically relevant experiments were performed in rats: prevention and treatment of ligature-induced periodontitis with RvE1 topical treatment. The gingival transcriptome was evaluated by RNA sequencing of mRNA. The composition of the subgingival microbiota was characterized by 16S rDNA sequencing. Periodontitis was assessed by bone morphometric measurements and histomorphometry of block sections. H&E and tartrate-resistant acid phosphatase staining were used to characterize and quantify inflammatory changes. RvE1 treatment prevented bone loss in ligature-induced periodontitis. Osteoclast density and inflammatory cell infiltration in the RvE1 groups were lower than those in the placebo group. RvE1 treatment reduced expression of inflammation-related genes, returning the expression profile to one more similar to health. Treatment of established periodontitis with RvE1 reversed bone loss, reversed inflammatory gene expression, and reduced osteoclast density. Assessment of the rat subgingival microbiota after RvE1 treatment revealed marked changes in both prevention and treatment experiments. The data suggest that modulation of local inflammation has a major role in shaping the composition of the subgingival microbiota.
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Disbiose/tratamento farmacológico , Ácido Eicosapentaenoico/análogos & derivados , Inflamação/tratamento farmacológico , Periodontite/tratamento farmacológico , Animais , Modelos Animais de Doenças , Ácido Eicosapentaenoico/administração & dosagem , Ácido Eicosapentaenoico/uso terapêutico , Inflamação/genética , Masculino , Ratos , Ratos WistarRESUMO
Different mechanisms have been hypothesized to explain the increase in prevalence and severity of periodontitis in older adults, including shifts in the periodontal microbiota. However, the actual impact of aging on the composition of subgingival biofilms remains unclear. In the present article, we provide an overview of the composition of the subgingival biofilm in older adults and the potential effects of age on the oral microbiome. In particular, this review covers the following topics: (i) the oral microbiota of an aging mouth; (ii) the effects of age and time on the human oral microbiome; (iii) the potential impact of inflammaging and immunosenescence in the host-oral microbiota interactions; and (iv) the relationship of the aging oral microbiota and Alzheimer's disease. Finally, we present analyses of data compiled from large clinical studies that evaluated the subgingival microbiota of periodontally healthy subjects and patients with periodontitis from a wide age spectrum (20-83 years of age).
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Envelhecimento/fisiologia , Biofilmes , Microbiota , Boca/microbiologia , Periodontite/microbiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/complicações , Bactérias/classificação , Bactérias/patogenicidade , Bolsa Gengival/microbiologia , Interações Hospedeiro-Patógeno/imunologia , Interações Hospedeiro-Patógeno/fisiologia , Humanos , Imunossenescência , Pessoa de Meia-Idade , Bolsa Periodontal/microbiologia , Periodontite/complicações , Periodonto/microbiologia , Fatores de TempoRESUMO
INTRODUCTION: The purpose of this study was to combine multiple displacement amplification and checkerboard DNA-DNA hybridization to qualitatively and quantitatively evaluate the microbiota present in infections refractory to endodontic treatment. METHODS: The subjects of this study were 40 patients presenting with periapical lesions refractory to endodontic treatment. Samples were taken by scraping or filing root canal walls with a #10 K-type hand file. Sample DNA was amplified by multiple displacement amplification, and the levels of 107 bacterial taxa were analyzed by checkerboard DNA-DNA hybridization. The taxa were divided into 3 distinct microbial populations depending on their mean proportion in samples (% DNA probe counts ± standard error of the mean) as follows: dominant (≥3.0%), subdominant (>1.6%-3.0%), and residual (≤1.6%) populations. The significance of differences was determined using the Mann-Whitney test. RESULTS: The taxa present with the highest mean proportions (constituting the dominant population) were Corynebacterium diphtheriae (8.03 ± 0.98), Porphyromonas gingivalis (5.42 ± 2.09), Streptococcus sobrinus (5.33 ± 0.69), and Stenotrophomonas maltophilia (4.72 ± 1.73). Among the subdominant population were Eubacterium saphenum (3.85 ± 1.06), Helicobacter pylori (3.16 ± 0.62), Dialister pneumosintes (3.12 ± 1.1), Clostridium difficile (2.74 ± 0.41), Enterobacter agglomerans (2.64 ± 0.54), Salmonella enterica (2.51 ± 0.52), Mobiluncus mulieris (2.44 ± 0.6), and Klebsiella oxytoca (2.32 ± 0.66). In the population of bacteria present at the lowest mean proportions (the residual population), Bacteroides ureolyticus (0.04 ± 0.01), Haemophilus influenzae (0.04 ± 0.02), and Prevotella oris (0.01 ± 0.01) were found at the lowest mean proportions. Enterococcus faecalis was detected in the residual population (0.52 ± 0.26). CONCLUSIONS: The microbial climax community in teeth refractory to endodontic treatment not only harbors medically important species but also contains distinct microbial consortia present with different population levels.
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Bactérias/isolamento & purificação , Cavidade Pulpar/microbiologia , Doenças Periapicais/microbiologia , Tratamento do Canal Radicular , Sondas de DNA , DNA Bacteriano/análise , Necrose da Polpa Dentária/microbiologia , Humanos , Microbiota , Hibridização de Ácido NucleicoRESUMO
AIM: The objective of this cross-sectional study was to examine the potential of peri-implant crevicular fluid (PICF) analytes to discriminate between peri-implant health and disease using a multi-biomarker approach. METHODS: We collected PICF samples from the mesio-buccal site of every implant (n = 145) from 52 subjects with peri-implantitis and measured the levels of 20 biomarkers using Luminex. We grouped implants and subjects based on the clinical characteristic of the sampled sites and implants into: healthy sites from healthy implants (HH), diseased sites from diseased implants (DD) and healthy sites from diseased implants (HD). The significance of the differences between the HH and DD groups was determined using general linear models controlling for false discovery rate. We used logistic regression to determine the best multi-biomarker models that could distinguish HH from DD subjects and HH from HD subjects. RESULTS: There were statistically significant differences between HH and DD groups for 12/20 biomarkers. Logistic regression resulted in a 6-biomarker model (Flt-3L, GM-CSF, IL-10, sCD40L, IL-17 and TNFα) that discriminated HH from DD subjects (AUC = 0.93) and a 3-biomarker model (IL-17, IL-1ra and vascular endothelial growth factor) that distinguished HH from DD subjects (AUC = 0.90). CONCLUSION: PICF biomarkers might help discriminate peri-implant health from disease.
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Líquido do Sulco Gengival , Biomarcadores , Estudos Transversais , Implantes Dentários , Humanos , Peri-Implantite , Fator A de Crescimento do Endotélio VascularRESUMO
AIM: The goal of this study was to identify progressing periodontal sites by applying linear mixed models (LMM) to longitudinal measurements of clinical attachment loss (CAL). METHODS: Ninety-three periodontally healthy and 236 periodontitis subjects had their CAL measured bi-monthly for 12 months. The proportions of sites demonstrating increases in CAL from baseline above specified thresholds were calculated for each visit. The proportions of sites reversing from the progressing state were also computed. LMM were fitted for each tooth site and the predicted CAL levels used to categorize sites regarding progression or regression. The threshold for progression was established based on the model-estimated error in predictions. RESULTS: Over 12 months, 21.2%, 2.8% and 0.3% of sites progressed, according to thresholds of 1, 2 and 3 mm of CAL increase. However, on average, 42.0%, 64.4% and 77.7% of progressing sites for the different thresholds reversed in subsequent visits. Conversely, 97.1%, 76.9% and 23.1% of sites classified as progressing using LMM had observed CAL increases above 1, 2 and 3 mm after 12 months, whereas mean rates of reversal were 10.6%, 30.2% and 53.0% respectively. CONCLUSION: LMM accounted for several sources of error in longitudinal CAL measurement, providing an improved method for classifying progressing sites.
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Doenças Periodontais , Progressão da Doença , Humanos , Estudos Longitudinais , Perda da Inserção Periodontal , Bolsa PeriodontalRESUMO
AIM: This exploratory randomized, controlled clinical trial sought to evaluate anti-inflammatory and -microbial effects of triclosan during experimental gingivitis as assessed by host response biomarkers and biofilm microbial pathogens. MATERIALS AND METHODS: Thirty participants were randomized to triclosan or control dentifrice groups who ceased homecare for 21 days in an experimental gingivitis (EG) protocol. Plaque and gingival indices and saliva, plaque, and gingival crevicular fluid (GCF) were assessed/collected at days 0, 14, 21 and 35. Levels and proportions of 40 bacterial species from plaque samples were determined using checkerboard DNA-DNA hybridization. Ten biomarkers associated with inflammation, matrix degradation, and host protection were measured from GCF and saliva and analysed using a multiplex array. Participants were stratified as "high" or "low" responders based on gingival index and GCF biomarkers and bacterial biofilm were combined to generate receiver operating characteristic curves and predict gingivitis susceptibility. RESULTS: No differences in mean PI and GI values were observed between groups and non-significant trends of reduction of host response biomarkers with triclosan treatment. Triclosan significantly reduced levels of A. actinomycetemcomitans and P. gingivalis during induction of gingivitis. CONCLUSIONS: Triclosan reduced microbial levels during gingivitis development (ClinicalTrials.gov NCT01799226).