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1.
Res Vet Sci ; 136: 396-407, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33799170

RESUMO

The purpose of the investigation was to determine whether canine gingival margin (GM) plaque is a reliable surrogate for subgingival (SG) plaque from a microbial community (microbiota) perspective. SG and GM plaque samples were collected from 381 dogs visiting pet hospitals in the USA, China and Thailand. Dogs with clinically healthy gingivae through to early periodontitis were included in the study. The samples were subject to next generation Illumina sequence analysis to allow microbiota comparisons to be made between the two plaque sources. Overall, the SG and GM samples indicated commonality via the majority community that were shared between them; health associations led to the identification of some significant taxa-specific differences. GM microbiota exhibited lower variability and diversity and were shown to reflect a sub-population of those associated with SG plaque. Both plaque niches, however, demonstrated similar changes in microbial signatures with health and early periodontal disease and did not indicate divergent trends. The key, most abundant microbiota of GM plaque strongly reflect those observed with SG plaque across health and early periodontitis. Microbiota in plaque from above the gum line may therefore be employed as a biomarker of oral health. This opens up the potential to use plaque, sampled from conscious dogs, to define oral health status and improve the diagnosis, treatments and interventions for periodontal disease.


Assuntos
Bactérias/isolamento & purificação , Placa Dentária/veterinária , Doenças do Cão/microbiologia , Gengiva/microbiologia , Microbiota , Doenças Periodontais/veterinária , Animais , Bactérias/genética , Biodiversidade , China , Estudos de Coortes , Placa Dentária/microbiologia , Cães , Feminino , Sequenciamento de Nucleotídeos em Larga Escala/veterinária , Masculino , Doenças Periodontais/diagnóstico , Doenças Periodontais/microbiologia , Tailândia
2.
Medicine (Baltimore) ; 100(10): e24924, 2021 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-33725852

RESUMO

ABSTRACT: Orthodontic treatment can lead to microbial-induced gingival inflammation and aseptic periodontal inflammations. The aim of this study was to investigate the relationship between salivary pro-inflammatory cytokines levels with gingival health status and oral microbe loads among patients undergoing orthodontic treatment.The present investigation was a cross-sectional study among a sample of 111 consecutive orthodontic patients (mean age 18.4 ±â€Š4.4 years). Clinical examinations were conducted to assess the gingival health status employing the Modified Gingival Index, Gingival Bleeding Index, and Plaque Index. Salivary microbiological assessments of total aerobic and anaerobic bacteria count, streptococci count, and lactobacilli count were undertaken. Saliva immunological assessments included Interleukin-1Beta (IL-1ß) and macrophage migration inhibitory factor (MIF) ELISA assays.The mean ±â€Šstandard deviation of salivary IL-1ß was 83.52 ±â€Š85.62 pg/ml and MIF was 4.12 ±â€Š0.96 ng/ml. Moderate positive correlations were found between salivary IL-1ß levels and total aerobic and anaerobic bacteria count, streptococci count, and lactobacilli count (r = 0.380-0.446, P < .001), and weak positive correlations between salivary MIF levels and total salivary aerobic and anaerobic bacteria counts (r = 0.249-0.306, P < .01) were observed. A positive correlation was found between salivary IL-1ß levels and Bleeding Index (r = 0.216, P < .05).The level of salivary IL-1ß positively correlates with oral bacterial load among orthodontic patients; the relationship between inflammatory cytokines and oral microflora deserved further study.


Assuntos
Gengivite/diagnóstico , Interleucina-1beta/análise , Aparelhos Ortodônticos/efeitos adversos , Saliva/química , Adolescente , Carga Bacteriana , Estudos Transversais , Feminino , Gengiva/imunologia , Gengiva/microbiologia , Gengivite/imunologia , Gengivite/microbiologia , Gengivite/prevenção & controle , Humanos , Interleucina-1beta/imunologia , Oxirredutases Intramoleculares/análise , Oxirredutases Intramoleculares/imunologia , Fatores Inibidores da Migração de Macrófagos/análise , Fatores Inibidores da Migração de Macrófagos/imunologia , Masculino , Microbiota/imunologia , Antissépticos Bucais/administração & dosagem , Adulto Jovem
3.
J Med Microbiol ; 70(3)2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33734952

RESUMO

Introduction. Oral tissues are generally homeostatic despite exposure to many potential inflammatory agents including the resident microbiota. This requires the balancing of inflammation by regulatory mechanisms and/or anti-inflammatory commensal bacteria. Thus, the levels of anti-inflammatory commensal bacteria in resident populations may be critical in maintaining this homeostatic balance.Hypothesis/Gap Statement. The incidence of immunosuppressive streptococci in the oral cavity is not well established. Determining the proportion of these organisms and the mechanisms involved may help to understand host-microbe homeostasis and inform development of probiotics or prebiotics in the maintenance of oral health.Aim. To determine the incidence and potential modes of action of immunosuppressive capacity in resident oral streptococci.Methodology. Supragingival plaque was collected from five healthy participants and supragingival and subgingival plaque from five with gingivitis. Twenty streptococci from each sample were co-cultured with epithelial cells±flagellin or LL-37. CXCL8 secretion was detected by ELISA, induction of cytotoxicity in human epithelial cells by lactate dehydrogenase release and NFκB-activation using a reporter cell line. Bacterial identification was achieved through partial 16S rRNA gene sequencing and next-generation sequencing.Results. CXCL8 secretion was inhibited by 94/300 isolates. Immunosuppressive isolates were detected in supragingival plaque from healthy (4/5) and gingivitis (4/5) samples, and in 2/5 subgingival (gingivitis) plaque samples. Most were Streptococcus mitis/oralis. Seventeen representative immunosuppressive isolates all inhibited NFκB activation. The immunosuppressive mechanism was strain specific, often mediated by ultra-violet light-labile factors, whilst bacterial viability was essential in certain species.Conclusion. Many streptococci isolated from plaque suppressed epithelial cell CXCL8 secretion, via inhibition of NFκB. This phenomenon may play an important role in oral host-microbe homeostasis.


Assuntos
Imunomodulação , Interleucina-8/metabolismo , Microbiota/imunologia , Boca/microbiologia , NF-kappa B/metabolismo , Streptococcus/imunologia , Células A549 , Linhagem Celular , Células Epiteliais/metabolismo , Gengiva/microbiologia , Gengivite/microbiologia , Humanos , Microbiota/genética , Streptococcus/classificação , Streptococcus/genética , Streptococcus/isolamento & purificação
4.
Methods Mol Biol ; 2210: 215-224, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32815142

RESUMO

Porphyromonas gingivalis is a major pathogen responsible for severe and chronic manifestations of periodontal disease, which is one of the most common infectious disorders of humans. Although human gingival epithelium prevents intrusions by periodontal bacteria, P. gingivalis is able to invade gingival epithelial cells. To study the dynamics and the fate of intracellular P. gingivalis, confocal laser scanning microscopy (CLSM) is a method of choice. Information gained with CLSM contains not only the number of P. gingivalis associated with gingival epithelial cells but also the bacterial localization on/inside the host cells, morphological change of host cells, and physical interaction between the bacteria and host organelle. In this chapter, we describe the protocols for microscopy techniques to morphologically study gingival epithelial cells infected by P. gingivalis.


Assuntos
Infecções por Bacteroidaceae/patologia , Células Epiteliais/patologia , Gengiva/patologia , Doenças Periodontais/patologia , Porphyromonas gingivalis/fisiologia , Infecções por Bacteroidaceae/microbiologia , Técnicas de Cultura de Células/métodos , Linhagem Celular , Células Epiteliais/microbiologia , Gengiva/citologia , Gengiva/microbiologia , Humanos , Microscopia Confocal/métodos , Doenças Periodontais/microbiologia , Porphyromonas gingivalis/isolamento & purificação , Coloração e Rotulagem/métodos
5.
Biochim Biophys Acta Mol Basis Dis ; 1867(1): 165991, 2021 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-33080346

RESUMO

Our previous study demonstrated that IL-10 secreting B (B10) cells alleviate inflammation and bone loss in experimental periodontitis. The purpose of this study is to determine whether antigen-specificity is required for the local infiltration of B10 cells. Experimental periodontitis was induced in the recipient mice by placement of silk ligature with or without the presence of live Porphyromonas gingivalis (P. gingivalis). Donor mice were pre-immunized by intraperitoneal (IP) injection of formalin-fixed P. gingivalis, or PBS as non-immunized control. Spleen B cells were purified and treated with LPS and CpG for 48 h to expand the B10 population in vitro. Fluorescence-labelled B10 cells were transferred into the recipient mice by tail vein injection and were tracked on day 0, 3, 5 and 10 using IVIS Spectrum in vivo imaging system. The number of B10 cells and P. gingivalis-binding B cells were significantly increased after in vitro treatment of LPS and CpG. On day 5, the fluorescence intensity in gingival tissues was the highest in mice transferred with B10 cells from pre-immunized donor mice. Gingival expression of IL-6, TNF-α, RANKL/OPG ratio and periodontal bone loss in recipient mice were significantly reduced, and the expression of IL-10 and the number of CD19+ B cells were significantly increased after pre-immunized B10 cell transfer in the presence of antigen, compared to those with non-immunized B10 cell transfer or no antigen presence. This study suggests that antigen specificity dictate the local infiltration of B10 cells into periodontal tissue and these antigen-specific B10 cells promote anti-inflammatory responses.


Assuntos
Antígenos de Bactérias/imunologia , Linfócitos B Reguladores/imunologia , Infecções por Bacteroidaceae , Gengiva , Periodontite , Porphyromonas gingivalis/imunologia , Animais , Infecções por Bacteroidaceae/diagnóstico por imagem , Infecções por Bacteroidaceae/imunologia , Infecções por Bacteroidaceae/microbiologia , Citocinas/imunologia , Diagnóstico por Imagem , Gengiva/diagnóstico por imagem , Gengiva/imunologia , Gengiva/microbiologia , Camundongos , Periodontite/diagnóstico por imagem , Periodontite/imunologia , Periodontite/microbiologia
6.
Periodontol 2000 ; 85(1): 82-89, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33226731

RESUMO

The microbial communities that inhabit the gingival crevice are responsible for the pathological processes that affect the periodontium. The changes in composition and function of subgingival bacteria as disease develops have been extensively studied. Subgingival communities, however, also contain fungi, Archaea, and viruses, which could contribute to the dysbiotic processes associated with periodontal diseases. High-throughput DNA sequencing has facilitated a better understanding of the mycobiome, archaeome, and virome. However, the number of studies available on the nonbacterial components of the subgingival microbiome remains limited in comparison with publications focusing on bacteria. Difficulties in characterizing fungal, archaeal, and viral populations arise from the small portion of the total metagenome mass they occupy and lack of comprehensive reference genome databases. In addition, specialized approaches potentially introducing bias are required to enrich for viral particles, while harsh methods of cell lysis are needed to recover nuclei acids from certain fungi. While the characterization of the subgingival diversity of fungi, Archaea and viruses is incomplete, emerging evidence suggests that they could contribute in different ways to subgingival dysbiosis. Certain fungi, such as Candida albicans are suggested to facilitate colonization of bacterial pathogens. Methanogenic Archaea are associated with periodontitis severity and are thought to partner synergistically with bacterial fermenters, while viruses may affect immune responses or shape microbial communities in ways incompletely understood. This review describes the manner in which omics approaches have improved our understanding of the diversity of fungi, Archaea, and viruses within subgingival communities. Further characterization of these understudied components of the subgingival microbiome is required, together with mechanistic studies to unravel their ecological role and potential contributions to dysbiosis.


Assuntos
Gengiva , Microbiota , Vírus , Archaea/genética , Bactérias/genética , DNA , Fungos , Gengiva/microbiologia , Humanos
7.
Sci Rep ; 10(1): 21989, 2020 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-33319834

RESUMO

Fixed orthodontic appliances are common and effective tools to treat malocclusion. Adverse effects of these appliances, such as dental caries and periodontitis, may be associated with alteration of the microbiome. This study investigated the impact of these appliances on the dynamics of the oral microbiome. Seventy-one patients were selected. Supragingival plaque samples were collected before placement (T0) and six months after placement (T1). Saliva samples were collected at T0 and T1, and then when appliance removal (T2). Microbial DNA was analyzed by 16S rRNA meta-sequencing. The diversity analysis indicated dynamic changes in the structure of the oral microbiome. Taxonomic analysis at phylum level showed a significant increase in Bacteroidetes and Saccharibacteria (formally TM7) and decrease in Proteobacteria and Actinobacteria over time, in both plaque and saliva. Genus level analysis of relative abundance indicated a significant increase in anaerobic and facultative anaerobes in both plaque and saliva. Fixed orthodontic appliances induced measurable changes in the oral microbiome. This was characterized by an increase in relative abundance of obligate anaerobes, including periodontal pathogens. It can be concluded that this dysbiosis induced by fixed orthodontic appliances is likely to represent a transitional stage in the shift in microbiome from healthy to periodontitis.


Assuntos
Grupo com Ancestrais do Continente Asiático , Microbiota , Boca/microbiologia , Aparelhos Ortodônticos Fixos/microbiologia , Bactérias/classificação , Biodiversidade , Gengiva/microbiologia , Humanos , Japão , Periodontite/microbiologia , Filogenia
8.
Med. oral patol. oral cir. bucal (Internet) ; 25(6): e745-e751, nov. 2020. graf, tab
Artigo em Inglês | IBECS | ID: ibc-197182

RESUMO

BACKGROUND: The objective of this cross-sectional clinical study was to analyze the differences in the microbiome in gingival sulci of adult patients in the presence or absence of chronic periodontitis. MATERIAL AND METHODS: Patients with or without periodontal disease were included in this cross-sectional study. Subgingival biofilm samples were collected and analyzed by 16S massive pyrosequencing. Functional analyses were also performed. RESULTS: A total of 15 phyla, 154 genera and 351 species were detected globally. Differences between disease and non-disease samples were observed in all taxonomical levels which suggest functional profile changes in the community. It was found that the main species associated with non-disease samples were reduced in disease but not completely suppressed. Analysis of the functional potential of the biofilms revealed a significantly higher activity related to endocytosis and phosphatidylinositol signaling in the disease group but lower cell adhesion molecules. CONCLUSIONS: Specific differences between health and disease suggest functional profile changes in the community, although bacteria associated with periodontal disease are also increased in health. Transcriptome studies should be conducted to confirm and deepen metabolic dysfunctions


No disponible


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Periodontite Crônica/microbiologia , Gengiva/microbiologia , Bactérias/isolamento & purificação , Estudos de Casos e Controles , Estudos Transversais , Sequenciamento de Nucleotídeos em Larga Escala , Microbiota , Reação em Cadeia da Polimerase , Bactérias/genética , RNA Ribossômico 16S/genética , Estatísticas não Paramétricas , RNA Bacteriano/isolamento & purificação
9.
Acta Odontol Latinoam ; 33(2): 104-111, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32920612

RESUMO

Candida dubliniensis (Cd) and Candida albicans (Ca) are the most frequently isolated yeasts in HIV+ patients. Some of the enzymes produced by these yeasts are considered virulence factors since they contribute to pathogenicity of Candida spp. The aim of the present study was to compare production of enzymes such as phospholipase (Ph), proteinase (P), and hemolysin (H) by Cd and Ca strains isolated from periodontal HIV-positive patients receiving and not receiving highly active antiretroviral therapy (HAART). Subgingival biofilm samples were obtained using paper points, and a sample of oral mucosa was taken using a swab. Phenotypic and molecular methods were used to isolate 39 strains of Candida, including 25 strains of Cd and 14 strains of Ca, obtained from 33 periodontal pocket samples and 6 oral mucosa samples collected from 15 HIV+ patients (8 receiving and 7 not receiving HAART). Malt egg-yolk agar, albumin agar and blood agar were used to evaluate pH, P and H production respectively. The strains were inoculated in duplicate and incubated at 37 ºC. Colony and halo diameters were measured. A greater proportion of Ca was observed in patients not receiving HAART, and a higher proportion of Cd was observed in those under HAART, Chi2 p< 0.001. Phospholipase production was observed in 92.9% percent of isolated Ca strains but in none of the isolated Cd strains. Proteinase production was high in Ca and Cd strains isolated from patients not receiving HAART. Hemolysin production was observed in all the studied strains, though it was significantly higher (p=0.04) in Ca and Cd strains isolated from patients not receiving HAART. To sum up, the proportion of Candida dubliniensis strains was highest in the subgingival biofilm of patients receiving HAART, and Cd strains were found to express fewer virulence factors than Ca strains.


Assuntos
Terapia Antirretroviral de Alta Atividade/métodos , Biofilmes/crescimento & desenvolvimento , Candida albicans/enzimologia , Candida albicans/isolamento & purificação , Candida/enzimologia , Candida/isolamento & purificação , Candidíase Bucal/microbiologia , Gengiva/microbiologia , Infecções por HIV/complicações , Candida/classificação , Candida/genética , Candida albicans/genética , Candidíase Bucal/complicações , Genótipo , Infecções por HIV/microbiologia , Humanos , Mucosa Bucal/microbiologia , Fenótipo , Reação em Cadeia da Polimerase , Fatores de Virulência/genética
10.
Sci Rep ; 10(1): 7823, 2020 05 08.
Artigo em Inglês | MEDLINE | ID: mdl-32385413

RESUMO

This study investigates the role of NLRP3 inflammasome and its main effector Caspase-1 in inflammation and alveolar bone resorption associated with periodontitis. Heat-killed Aggregatibacter actinomycetemcomitans (Aa) was injected 3x/week (4 weeks) into gingival tissues of wild-type (WT), Nlrp3-KO and Caspase1-KO mice. Bone resorption was measured by µCT and osteoclast number was determined by tartrate-resistant acid phosphatase (TRAP) staining. Inflammation was assessed histologically (H/E staining and immunofluorescence of CD45 and Ly6G). In vitro studies determined the influence of Nlrp3 and Caspase-1 in Rankl-induced osteoclast differentiation and activity and on LPS-induced expression of inflammation-associated genes. Bone resorption was significantly reduced in Casp1-KO but not in Nlrp3-KO mice. Casp1-KO mice had increased in osteoclast numbers, whereas the inflammatory infiltrate or on gene expression were similar to those of WT and Nlrp3-KO mice. Strikingly, osteoclasts differentiated from Nlrp3-deficient macrophages had increased resorbing activity in vitro. LPS-induced expression of Il-10, Il-12 and Tnf-α was significantly reduced in Nlrp3- and Casp1-deficient macrophages. As an inceptive study, these results suggest that Nlrp3 inflammasome does not play a significant role in inflammation and bone resorption in vivo and that Caspase-1 has a pro-resorptive role in experimental periodontal disease.


Assuntos
Perda do Osso Alveolar/genética , Caspase 1/genética , Inflamação/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Periodontite/genética , Aggregatibacter actinomycetemcomitans , Perda do Osso Alveolar/microbiologia , Perda do Osso Alveolar/patologia , Animais , Diferenciação Celular/efeitos dos fármacos , Modelos Animais de Doenças , Gengiva/crescimento & desenvolvimento , Gengiva/microbiologia , Humanos , Inflamação/microbiologia , Inflamação/patologia , Interleucina-10/genética , Interleucina-12/genética , Camundongos , Camundongos Knockout , Osteoclastos/microbiologia , Osteoclastos/patologia , Periodontite/microbiologia , Periodontite/patologia , Ligante RANK/genética , Fator de Necrose Tumoral alfa/genética
11.
PLoS One ; 15(5): e0232912, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32392236

RESUMO

The study of oral disease progression, in relation to the accumulation of subgingival biofilm in gingivitis and periodontitis is limited, due to either the ability to monitor plaque in vitro. When compared, optical spectroscopic techniques offer advantages over traditional destructive or biofilm staining approaches, making it a suitable alternative for the analysis and continued development of three-dimensional structures. In this work, we have developed a confocal Raman spectroscopy analysis approach towards in vitro subgingival plaque models. The main objective of this study was to develop a method for differentiating multiple oral subgingival bacterial species in planktonic and biofilm conditions, using confocal Raman microscopy. Five common subgingival bacteria (Fusobacterium nucleatum, Streptococcus mutans, Veillonella dispar, Actinomyces naeslundii and Prevotella nigrescens) were used and differentiated using a 2-way orthogonal Partial Least Square with Discriminant Analysis (O2PLS-DA) for the collected spectral data. In addition to planktonic growth, mono-species biofilms cultured using the 'Zürich Model' were also analyzed. The developed method was successfully used to predict planktonic and mono-species biofilm species in a cross validation setup. The results show differences in the presence and absence of chemical bands within the Raman spectra. The O2PLS-DA model was able to successfully predict 100% of all tested planktonic samples and 90% of all mono-species biofilm samples. Using this approach we have shown that Confocal Raman microscopy can analyse and predict the identity of planktonic and mono-species biofilm species, thus enabling its potential as a technique to map oral multi-species biofilm models.


Assuntos
Bactérias/isolamento & purificação , Gengivite/microbiologia , Microscopia Óptica não Linear/métodos , Periodontite/microbiologia , Actinomyces , Técnicas Bacteriológicas/métodos , Biofilmes/crescimento & desenvolvimento , Meios de Cultura , Fusobacterium nucleatum , Gengiva/microbiologia , Viabilidade Microbiana , Microbiota , Microscopia Confocal/métodos , Plâncton , Prevotella intermedia , Streptococcus mutans , Veillonella
12.
Arch Oral Biol ; 115: 104742, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32416352

RESUMO

BACKGROUND AND OBJECTIVE: Periodontitis is an oral chronic inflammatory disease caused by dental plaque. It is comorbid with numerous systemic diseases and associated with several predisposing factors, such as chronic kidney disease (CKD). Peritoneal dialysis is one of the ultimate treatments for patients with severe CKD. However, peritoneal dialysis patients with periodontitis often will be accompanied with more poor oral hygiene and periodontal clinical indexes. This study aimed to compare the microbial flora of periodontitis patients with or without peritoneal dialysis. METHODS: Sixteen peritoneal dialysis patients with periodontitis (P group) and 16 patients with periodontitis only (C group) were selected. Subgingival plaque samples of them were processed for bioinformatics analysis by 16S rDNA gene sequencing. RESULTS: The diversity indices and species richness in the P group were insignificantly higher than that in the C group (P > 0.05). The two groups exhibited different microbial community structure. At Genus level, Prevotellaceae, Selenomonas, Aggregatibacter, Anaeroglobus, TM7_[G-5], and Centipeda were significantly enriched in the P group than those in the C group. CONCLUSIONS: This study demonstrated that specific microbes enriched in the subgingival flora of peritoneal dialysis patients with periodontitis.


Assuntos
Microbiota , Periodontite , Diálise Peritoneal , Bactérias/genética , Gengiva/microbiologia , Humanos , Periodontite/microbiologia , RNA Ribossômico 16S/genética
13.
Arch Oral Biol ; 116: 104766, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32470832

RESUMO

OBJECTIVE: Diabetes increases the incidence/severity of periodontal diseases by inducing a chronic inflammation, driven by accumulation of AGEs (advanced glycation end products). We tested whether glycated human serum albumin (G-HSA, a form of AGE), representing a diabetic state, augments the pro-inflammatory response of human gingival fibroblasts (hGFs) to a bacterial challenge (Porphyromonas gingivalis Lipopolysaccharide (LPS)). METHODS: Primary hGFs were incubated with LPS (0.5-5 µg/mL) and G-HSA (50-200 µg/mL) and the production and gene expression of IL-1ß, IL-6, IL-8, MMP-1, MCP-1, and TNFα were analyzed by Magnetic Luminex Assay and real-time PCR, respectively. Non-glycated serum albumin (HSA) served as negative control. Cytotoxicity of the 2 agents was tested with an XTT assay. NFκB activation (p65 phosphorylation) was measured with an ELISA. RESULTS: P. gingivalis LPS and G-HSA were not toxic to hGFs and increased the amount of MMP-1, MCP-1, IL-6, and IL-8, (but not TNFα and IL-1ß) secreted into the medium at 24 h. Control HSA had no effect. Many LPS/G-HSA combinations displayed a synergistic stimulation of these molecules. Both agents increased mRNA levels of these 4 molecules at 6 h, 12 h or both (IL-6). NFκB activation at 6 h was caused by both agents with a possible synergism at the higher concentrations. CONCLUSIONS: glycated albumin augments the pro-inflammatory response of human gingival fibroblasts to P. gingivalis LPS. Thus, AGE accumulation in diabetes could aggravate periodontal inflammation by augmenting the pro-inflammatory response of host GFs to P. gingivalis, a well-recognized periopathogenic bacteria.


Assuntos
Gengiva , Lipopolissacarídeos , Porphyromonas gingivalis , Albumina Sérica , Células Cultivadas , Fibroblastos , Gengiva/microbiologia , Humanos , Interleucina-6 , NF-kappa B
14.
Infect Immun ; 88(5)2020 04 20.
Artigo em Inglês | MEDLINE | ID: mdl-32041789

RESUMO

Programmed death-ligand 1 (PD-L1/B7-H1) serves as a cosignaling molecule in cell-mediated immune responses and contributes to chronicity of inflammation and the escape of tumor cells from immunosurveillance. Here, we investigated the molecular mechanisms leading to PD-L1 upregulation in human oral carcinoma cells and in primary human gingival keratinocytes in response to infection with Porphyromonas gingivalis (P. gingivalis), a keystone pathogen for the development of periodontitis. The bacterial cell wall component peptidoglycan uses bacterial outer membrane vesicles to be taken up by cells. Internalized peptidoglycan triggers cytosolic receptors to induce PD-L1 expression in a myeloid differentiation primary response 88 (Myd88)-independent and receptor-interacting serine/threonine-protein kinase 2 (RIP2)-dependent fashion. Interference with the kinase activity of RIP2 or mitogen-activated protein (MAP) kinases interferes with inducible PD-L1 expression.


Assuntos
Antígeno B7-H1/metabolismo , Infecções por Bacteroidaceae/metabolismo , Carcinoma/metabolismo , Parede Celular/metabolismo , Neoplasias Bucais/metabolismo , Porphyromonas gingivalis/metabolismo , Proteína Serina-Treonina Quinase 2 de Interação com Receptor/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Infecções por Bacteroidaceae/microbiologia , Carcinoma/microbiologia , Linhagem Celular Tumoral , Gengiva/metabolismo , Gengiva/microbiologia , Humanos , Queratinócitos/metabolismo , Queratinócitos/microbiologia , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Neoplasias Bucais/microbiologia , Periodontite/metabolismo , Periodontite/microbiologia , Regulação para Cima/fisiologia
15.
BMC Oral Health ; 20(1): 27, 2020 01 30.
Artigo em Inglês | MEDLINE | ID: mdl-32000757

RESUMO

BACKGROUND: Both substance P and hypoxia-inducible factor 1 alpha (HIF-1α) are involved in inflammation and angiogenesis. However, the relationship between substance P and HIF-1α in rat periodontitis is still unknown. METHODS: Ligation-induced rat periodontitis was established to observe the distribution and expression of substance P and HIF-1α by immunohistochemistry. Rat gingival fibroblasts were cultured and stimulated with Porphyromonas gingivalis lipopolysaccharide (LPS). Recombinant substance P was applied to elaborate the relationship between substance P and HIF-1α in gingival fibroblasts in vitro. Primary mouse bone marrow-derived macrophages (BMMs) were isolated and cultured to observe the effect of substance P on receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis by TRAP staining. Western blotting was used to investigate the expression of HIF-1α, osteoprotegerin (OPG) and RANKL. RESULTS: Rat experimental periodontitis was successfully established 6 weeks after ligation. Gingival inflammatory infiltration and alveolar bone loss were observed. Positive expression of substance P was found in the infiltrating cells. Higher HIF-1α levels were observed in periodontitis compared to that of normal tissues. Substance P upregulated the level of HIF-1α in gingival fibroblasts with or without 1 µg/ml LPS in vitro (*P < 0.05). Substance P upregulated the expression of HIF-1α in RANKL-stimulated BMMs in vitro. Substance P also increased the RANKL/OPG ratio in gingival fibroblasts (*P < 0.05). Both 10 nM and 50 nM substance P promoted RANKL-induced osteoclast differentiation (*P < 0.05). CONCLUSION: Substance P participates in periodontitis by upregulating HIF-1α and the RANKL/OPG ratio.


Assuntos
Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Osteoprotegerina/genética , Periodontite/metabolismo , Porphyromonas gingivalis/isolamento & purificação , Ligante RANK/genética , Substância P/genética , Animais , Regulação da Expressão Gênica , Gengiva/microbiologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Masculino , Camundongos , Osteoclastos , Osteoprotegerina/metabolismo , Periodontite/microbiologia , Porphyromonas gingivalis/genética , Ratos , Ratos Wistar , Substância P/metabolismo , Regulação para Cima/genética
16.
Sci Rep ; 10(1): 3008, 2020 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-32080300

RESUMO

Several proteins and peptides in saliva were shown to stimulate gingival wound repair, but the role of salivary metabolites in this process remains unexplored. In vitro gingival re-epithelialization kinetics were determined using unstimulated saliva samples from healthy individuals collected during an experimental gingivitis study. Elastic net regression with stability selection identified a specific metabolite signature in a training dataset that was associated with the observed re-epithelialization kinetics and enabled its prediction for all saliva samples obtained in the clinical study. This signature encompassed ten metabolites, including plasmalogens, diacylglycerol and amino acid derivatives, which reflect enhanced host-microbe interactions. This association is in agreement with the positive correlation of the metabolite signature with the individual's gingival bleeding index. Remarkably, intra-individual signature-variation over time was associated with elevated risk for gingivitis development. Unravelling how these metabolites stimulate wound repair could provide novel avenues towards therapeutic approaches in patients with impaired wound healing capacity.


Assuntos
Eritritol/uso terapêutico , Gengiva/efeitos dos fármacos , Gengivite/metabolismo , Hemorragia/metabolismo , Metaboloma , Saliva/metabolismo , Adolescente , Adulto , Aminoácidos/metabolismo , Aminoácidos/farmacologia , Bioensaio , Estudos de Casos e Controles , Linhagem Celular , Diglicerídeos/metabolismo , Diglicerídeos/farmacologia , Suscetibilidade a Doenças , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Feminino , Gengiva/metabolismo , Gengiva/microbiologia , Gengiva/patologia , Gengivite/tratamento farmacológico , Gengivite/microbiologia , Gengivite/patologia , Hemorragia/tratamento farmacológico , Hemorragia/microbiologia , Hemorragia/patologia , Interações Hospedeiro-Patógeno , Humanos , Masculino , Pessoa de Meia-Idade , Plasmalogênios/metabolismo , Plasmalogênios/farmacologia , Reepitelização/efeitos dos fármacos , Reepitelização/fisiologia , Saliva/química , Saliva/microbiologia , Índice de Gravidade de Doença , Streptococcus mutans/crescimento & desenvolvimento , Streptococcus mutans/patogenicidade
17.
Arch Biochem Biophys ; 682: 108278, 2020 03 30.
Artigo em Inglês | MEDLINE | ID: mdl-31981541

RESUMO

Oral microbes are a contributing factor to hyperglycemia by inducing an increase in insulin resistance resulting in uncontrolled blood glucose levels. However, the relationship between the distribution of oral flora and hyperglycemia is still controversial. Combining the power of MALDI-Biotyper with anaerobic bacterial culture, this study explores the correlation between anaerobic bacteria in the oral cavity and blood glucose levels. The results demonstrated that altered blood glucose levels contributed to a varied bacterial distribution in the oral cavity. Specifically, Veillonella spp. and Prevotella spp. were identified in a higher proportion in people with elevated blood glucose levels. Six bacterial species identified in this study (Prevotella melaninogenica, Campylobacter rectus, Streptococcus gordonii, Streptococcus mitis, Streptococcus salivarius, and Veillonella parvula) not only demonstrated a positive association with higher blood glucose levels, but also likely contribute to the development of the condition. The data demonstrated MALDI-TOF MS to be a simpler, faster, and more economical clinical identification tool that provides clarity and depth to the research on blood glucose and oral microbiota.


Assuntos
Gengiva/microbiologia , Hiperglicemia/microbiologia , Microbiota , Saliva/microbiologia , Adulto , Idoso , Bactérias Anaeróbias , Glicemia/análise , Campylobacter rectus , Feminino , Hemoglobina A Glicada/análise , Humanos , Masculino , Pessoa de Meia-Idade , Prevotella/metabolismo , Prevotella melaninogenica , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Streptococcus gordonii , Streptococcus mitis , Streptococcus salivarius , Veillonella/metabolismo
18.
J Appl Oral Sci ; 28: e20190266, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31800877

RESUMO

OBJECTIVE: The microbial composition of pericoronitis (Pc) is still controversial; it is not yet clear if the microbial profile of these lesions is similar to the profile observed in periodontitis (Pd). Therefore, the aim of the present study was to describe the microbial profile of Pc lesions and compare it directly with that of subjects with Pd. METHODOLOGY: Subjects with Pc and Pd were selected, and subgingival biofilm samples were collected from (i) third molars with symptomatic Pc (Pc-T), (ii) contralateral third molars without Pc (Pc-C) and (iii) teeth with a probing depth >3 mm from subjects with Pd. Counts and proportions of 40 bacterial species were evaluated using a checkerboard DNA-DNA hybridization technique. RESULTS: Twenty-six patients with Pc and 18 with Pd were included in the study. In general, higher levels of microorganisms were observed in Pd. Only Actinomyces oris and Eubacterium nodatum were present in higher mean counts in the Pc-T group in comparison with the Pc-C and Pd-C groups (p<0.05). The microbiota associated with Pc-T was similar to that found in Pc-C. Sites with Pc lesions had lower proportions of red complex in comparison with the Pd sites. CONCLUSION: The microbiota of Pc is very diverse, but these lesions harbour lower levels of periodontal pathogens than Pd.


Assuntos
Bactérias/isolamento & purificação , Pericoronite/microbiologia , Periodontite/microbiologia , Análise por Ativação , Adulto , Idoso , Carga Bacteriana , Biofilmes , Estudos Transversais , Sondas de DNA , Feminino , Gengiva/microbiologia , Humanos , Masculino , Pessoa de Meia-Idade , Valores de Referência , Adulto Jovem
19.
Gene ; 727: 144258, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31759984

RESUMO

Peri-implantitis is similar to periodontitis in both symptoms and treatment; however, their level of similarity remains controversial. Here, we compared multiple cases of periodontitis and peri-implantitis through transcriptome and methylome profiling, and analyzed the effects of smoking as a typical risk factor. Human gingival tissues were obtained from 20 patients with periodontitis or peri-implantitis via periodontal surgical procedures. Total RNA and genomic DNA were isolated, and transcriptome and methylome datasets were generated. Comprehensive analysis of differential gene expression, DNA methylation, and protein-protein interactions indicated that periodontitis and peri-implantitis share biological similarities; however, hierarchical clustering between the two disease groups revealed distinct molecular characteristics. These differences might be related to structural differences in natural tooth-bone and implant-bone. Additionally, smoking differentially affected periodontitis and peri-implantitis in terms of host-defense mechanism impairment. Within the limitations of this study, the results provide evidence that peri-implantitis is distinct from periodontitis and that smoking potentially affects disease progression. Our study provides a foundation for the rational design of a large-scale study in the future for a more comprehensive analysis that includes microbiome and clinical data.


Assuntos
Peri-Implantite/genética , Periodontite/genética , Epigenoma/genética , Feminino , Perfilação da Expressão Gênica/métodos , Gengiva/microbiologia , Humanos , Masculino , Microbiota/genética , Pessoa de Meia-Idade , Peri-Implantite/metabolismo , Fatores de Risco , Fumar , Uso de Tabaco/efeitos adversos , Uso de Tabaco/genética , Transcriptoma/genética
20.
ISME J ; 14(2): 519-530, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31673077

RESUMO

Type 2 diabetes mellitus (T2DM) is a systemic disease, predisposing patients to other inflammatory conditions including periodontitis. The subgingival microbiome, a key player in periodontitis pathogenesis, is not well characterized in T2DM population. To better understand whether the subgingival microbiome is different between T2DM and systemically healthy, nondiabetic (ND) subjects, we performed a longitudinal analysis of the subgingival microbiome in T2DM patients (n = 15) compared with ND subjects (n = 16). Using metagenomic shotgun sequencing, we investigated the microbiome in the healthy periodontal state, periodontitis state, and resolved state after treatment. We found that in the periodontitis state, the shift in the subgingival microbiome from the healthy state was less prominent in T2DM compared with ND subjects, yet the clinical signs of disease were similar for both. Furthermore, we revealed highly correlated presence of pathogenic species in relative abundance not only in the periodontitis state, but also in the healthy state in T2DM, suggesting an elevated risk of progression to periodontitis in this cohort. We further investigated the functional potentials of the subgingival microbiome and identified a set of microbial marker genes associated with the clinical states. These genes were significantly enriched in 21 pathways, some of which are associated with periodontitis and some potentially link T2DM and periodontitis. This study identified the longitudinal changes of the subgingival microbiome associated with periodontitis in T2DM and suggests that T2DM patients are more susceptible to shifts in the subgingival microbiome toward dysbiosis, potentially due to impaired host metabolic and immune regulation.


Assuntos
Diabetes Mellitus Tipo 2/complicações , Metagenoma , Microbiota/genética , Periodontite/microbiologia , Adulto , Bactérias/classificação , Bactérias/genética , Estudos de Casos e Controles , Coinfecção/microbiologia , Disbiose/genética , Feminino , Gengiva/microbiologia , Doenças da Gengiva/microbiologia , Humanos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Boca/microbiologia
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