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1.
Front Cell Infect Microbiol ; 11: 722499, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34722331

RESUMO

Leucocyte- and platelet rich fibrin (L-PRF) is an autologous biomaterial used in regenerative procedures. It has an antimicrobial activity against P. gingivalis although the mechanism is not fully understood. It was hypothesized that L-PRF exudate releases hydrogen peroxide and antimicrobial peptides that inhibit P. gingivalis growth. Agar plate and planktonic culture experiments showed that the antimicrobial effect of L-PRF exudate against P. gingivalis was supressed by peroxidase or pepsin exposure. In developing multi-species biofilms, the antimicrobial effect of L-PRF exudate was blocked only by peroxidase, increasing P. gingivalis growth with 1.3 log genome equivalents. However, no effect was shown on other bacteria. Pre-formed multi-species biofilm trials showed no antimicrobial effect of L-PRF exudate against P. gingivalis or other species. Our findings showed that L-PRF exudate may release peroxide and peptides, which may be responsible for its antimicrobial effect against P. gingivalis. In addition, L-PRF exudate had an antimicrobial effect against P. gingivalis in an in vitro developing multi-species biofilm.


Assuntos
Anti-Infecciosos , Fibrina Rica em Plaquetas , Anti-Infecciosos/farmacologia , Biofilmes , Exsudatos e Transudatos , Projetos Piloto , Plâncton , Porphyromonas gingivalis
2.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 52(5): 754-758, 2021 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-34622588

RESUMO

Objective: To explore the effect of polystyrene (PS) and PS-polyvinylpyrrolidone (PVP) electrospun materials on the adhesion ability of Porphyromonas gingivalis( P. gingivalis), a common periodontal pathogen. Methods: PS and PS-PVP electrospun materials were prepared with stainless steel needles in high-voltage electric field. The growth and adhesion of P. gingivalis on the surface of different materials were observed with scanning electron microscope (SEM). The changes in the amount of P. gingivalis biofilm formed on the surface of different materials were measured according to viable colony forming units (CFU). The effect of surface charge of the different materials on the adhesion ability of P. gingivalis was determined through changing the charge properties on the surface of the electrospun materials. Results: SEM images showed that both PS and PS-PVP can be used to form electrospun fibers with a diameter of 0.2 µm. SEM images and CFU counts of the biofilm at 24 h and 48 h showed that there was a smaller amount of P. gingivalis biofilm on the surface of the two materials ( P<0.05). After treatment with tetrabutylammonium bromide (TBAB), the surface charge of the PS-PVP electrospun material changed from being negatively charged to being positively charged, and the amount of bacterial adhesion on the surface increased significantly in comparison to that of untreated PS and PS-PVP materials ( P<0.05). Conclusion: PS and PS-PVP electrospun materials can be used to reduce the adhesion ability of P. gingivalis on the surface of different materials, and this ability may be related to the surface charge properties of the materials.


Assuntos
Porphyromonas gingivalis , Povidona , Biofilmes , Fibras na Dieta , Poliestirenos , Povidona/farmacologia
3.
Int J Oral Sci ; 13(1): 31, 2021 09 30.
Artigo em Inglês | MEDLINE | ID: mdl-34593756

RESUMO

Ulcerative Colitis (UC) has been reported to be related to Porphyromonas gingivalis (P. gingivalis). Porphyromonas gingivalis peptidylarginine deiminase (PPAD), a virulence factor released by P. gingivalis, is known to induce inflammatory responses. To explore the pathological relationships between PPAD and UC, we used homologous recombination technology to construct a P. gingivalis strain in which the PPAD gene was deleted (Δppad) and a Δppad strain in which the PPAD gene was restored (comΔppad). C57BL/6 mice were orally gavaged with saline, P. gingivalis, Δppad, or comΔppad twice a week for the entire 40 days (days 0-40), and then, UC was induced by dextran sodium sulfate (DSS) solution for 10 days (days 31-40). P. gingivalis and comΔppad exacerbated DDS-induced colitis, which was determined by assessing the parameters of colon length, disease activity index, and histological activity index, but Δppad failed to exacerbate DDS-induced colitis. Flow cytometry and ELISA revealed that compared with Δppad, P. gingivalis, and comΔppad increased T helper 17 (Th17) cell numbers and interleukin (IL)-17 production but decreased regulatory T cells (Tregs) numbers and IL-10 production in the spleens of mice with UC. We also cocultured P. gingivalis, Δppad, or comΔppad with T lymphocytes in vitro and found that P. gingivalis and comΔppad significantly increased Th17 cell numbers and decreased Treg cell numbers. Immunofluorescence staining of colon tissue paraffin sections also confirmed these results. The results suggested that P. gingivalis exacerbated the severity of UC in part via PPAD.


Assuntos
Colite Ulcerativa , Porphyromonas gingivalis , Desiminases de Arginina em Proteínas , Animais , Colite Ulcerativa/microbiologia , Camundongos , Camundongos Endogâmicos C57BL , Porphyromonas gingivalis/enzimologia , Porphyromonas gingivalis/patogenicidade , Fatores de Virulência
4.
Medicina (Kaunas) ; 57(10)2021 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-34684091

RESUMO

Background and Objectives: Periodontal disease is a chronic inflammatory disease in which gradual destruction of tissues around teeth is caused by plaque formed by pathogenic bacteria. The purpose of this study was to evaluate the potential of 75% ethanol extract of Colocasia antiquorum var. esculenta (CA) as a prophylactic and improvement agent for periodontal disease in vitro and in vivo. Materials and Methods: The antimicrobial efficacy of CA against Porphyromonas gingivalis (P. gingivalis, ATCC 33277) was evaluated using minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) test, and cytotoxicity was confirmed by CCK-8 assay. For the in vivo study, P. gingivalis was applied by oral gavage to BALB/c mice. Forty-two days after the first inoculation of P. gingivalis, intraoral swabs were taken for microbiome analysis, and the mice were sacrificed to evaluate the alveolar bone loss. Results: The MIC of CA against P. gingivalis was 31.3 µg/mL, the MBC was 62.5 µg/mL, with no cytotoxicity. The diversity of the oral microbiome decreased in the positive control group, while those of the VA (varnish) and VCA (varnish added with CA) groups increased as much as in the negative control group, although the alveolar bone loss was not induced in the mouse model. Conclusions: CA showed antibacterial effects in vitro, and the VA and VCA groups exhibited increased diversity in the oral microbiome, suggesting that CA has potential for improving periodontal disease.


Assuntos
Colocasia , Doenças Periodontais , Animais , Camundongos , Camundongos Endogâmicos BALB C , Doenças Periodontais/tratamento farmacológico , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Porphyromonas gingivalis
5.
Front Cell Infect Microbiol ; 11: 723821, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34616690

RESUMO

Ancient dental calculus, formed from dental plaque, is a rich source of ancient DNA and can provide information regarding the food and oral microbiology at that time. Genomic analysis of dental calculus from Neanderthals has revealed the difference in bacterial composition of oral microbiome between Neanderthals and modern humans. There are few reports investigating whether the pathogenic bacteria of periodontitis, a polymicrobial disease induced in response to the accumulation of dental plaque, were different between ancient and modern humans. This study aimed to compare the bacterial composition of the oral microbiome in ancient and modern human samples and to investigate whether lifestyle differences depending on the era have altered the bacterial composition of the oral microbiome and the causative bacteria of periodontitis. Additionally, we introduce a novel diagnostic approach for periodontitis in ancient skeletons using micro-computed tomography. Ancient 16S rDNA sequences were obtained from 12 samples at the Unko-in site (18th-19th century) of the Edo era (1603-1867), a characteristic period in Japan when immigrants were not accepted. Furthermore, modern 16S rDNA data from 53 samples were obtained from a database to compare the modern and ancient microbiome. The microbial co-occurrence network was analyzed based on 16S rDNA read abundance. Eubacterium species, Mollicutes species, and Treponema socranskii were the core species in the Edo co-occurrence network. The co-occurrence relationship between Actinomyces oricola and Eggerthella lenta appeared to have played a key role in causing periodontitis in the Edo era. However, Porphyromonas gingivalis, Fusobacterium nucleatum subsp. vincentii, and Prevotella pleuritidis were the core and highly abundant species in the co-occurrence network of modern samples. These results suggest the possibility of differences in the pathogens causing periodontitis during different eras in history.


Assuntos
Bactérias/classificação , Periodontite , Actinobacteria , Actinomyces , Fusobacterium , História do Século XVII , História do Século XVIII , História do Século XIX , Humanos , Japão , Periodontite/diagnóstico , Periodontite/história , Periodontite/microbiologia , Porphyromonas gingivalis , Prevotella , Treponema , Microtomografia por Raio-X
6.
Front Cell Infect Microbiol ; 11: 727732, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34692561

RESUMO

The development of periodontitis is associated with an imbalanced subgingival microbial community enriched with species such as the traditionally classified red-complex bacteria (Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola). Saliva has been suggested as an alternative to subgingival plaque for the microbial analysis due to its easy and non-invasive collection. This systematic review aims to determine whether the levels of red-complex bacteria assessed using saliva reflect those in subgingival plaque from periodontitis patients. The MEDLINE, EMBASE, and Cochrane Library databases were searched up to April 30, 2021. Studies were considered eligible if microbial data of at least one of the red-complex species were reported in both saliva and subgingival plaque from periodontitis patients, based on DNA-based methods. Of the 17 included studies, 4 studies used 16S rRNA gene sequencing techniques, and the rest used PCR-based approaches. The detection frequency of each red-complex species in periodontitis patients was reported to be > 60% in most studies, irrespective of samples types. Meta-analyses revealed that both detection frequencies and relative abundances of red-complex bacteria in saliva were significantly lower than those in subgingival plaque. Moreover, the relative abundances of all 3 bacterial species in saliva showed significantly positive correlation with those in subgingival plaque. In conclusion, current evidence suggests that one-time saliva sampling cannot replace subgingival plaque for microbial analysis of the red-complex bacteria in periodontitis patients. Given the positive microbial associations between saliva and subgingival plaque, a thorough review of longitudinal clinical studies is needed to further assess the role of saliva.


Assuntos
Periodontite , Saliva , Aggregatibacter actinomycetemcomitans , Humanos , Porphyromonas gingivalis/genética , RNA Ribossômico 16S/genética , Treponema denticola/genética
7.
Am J Physiol Heart Circ Physiol ; 321(5): H948-H962, 2021 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-34597184

RESUMO

Oral and gum health have long been associated with incidence and outcomes of cardiovascular disease. Periodontal disease increases myocardial infarction (MI) mortality by sevenfold through mechanisms that are not fully understood. The goal of this study was to evaluate whether lipopolysaccharide (LPS) from a periodontal pathogen accelerates inflammation after MI through memory T-cell activation. We compared four groups [no MI, chronic LPS, day 1 after MI, and day 1 after MI with chronic LPS (LPS + MI); n = 68 mice] using the mouse heart attack research tool 1.0 database and tissue bank coupled with new analyses and experiments. LPS + MI increased total CD8+ T cells in the left ventricle versus the other groups (P < 0.05 vs. all). Memory CD8+ T cells (CD44 + CD27+) were 10-fold greater in LPS + MI than in MI alone (P = 0.02). Interleukin (IL)-4 stimulated splenic CD8+ T cells away from an effector phenotype and toward a memory phenotype, inducing secretion of factors associated with the Wnt/ß-catenin signaling that promoted monocyte migration and decreased viability. To dissect the effect of CD8+ T cells after MI, we administered a major histocompatibility complex-I-blocking antibody starting 7 days before MI, which prevented effector CD8+ T-cell activation without affecting the memory response. The reduction in effector cells diminished infarct wall thinning but had no effect on macrophage numbers or MertK expression. LPS + MI + IgG attenuated macrophages within the infarct without effecting CD8+ T cells, suggesting these two processes were independent. Overall, our data indicate that effector and memory CD8+ T cells at post-MI day 1 are amplified by chronic LPS to potentially promote infarct wall thinning.NEW & NOTEWORTHY Although there is a well-documented link between periodontal disease and heart health, the mechanisms are unclear. Our study indicates that in response to circulating periodontal endotoxins, memory CD8+ T cells are activated, resulting in an acceleration of macrophage-mediated inflammation after MI. Blocking activation of effector CD8+ T cells had no effect on the macrophage numbers or wall thinning at post-MI day 1, indicating that this response was likely due in part to memory CD8+ T cells.


Assuntos
Linfócitos T CD8-Positivos/imunologia , Memória Imunológica , Lipopolissacarídeos , Ativação Linfocitária , Infarto do Miocárdio/imunologia , Miocárdio/imunologia , Periodontite/imunologia , Porphyromonas gingivalis , Cicatrização , Animais , Linfócitos T CD8-Positivos/metabolismo , Células Cultivadas , Modelos Animais de Doenças , Feminino , Mediadores da Inflamação/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/metabolismo , Masculino , Camundongos , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/patologia , Miocárdio/metabolismo , Miocárdio/patologia , Periodontite/induzido quimicamente , Periodontite/metabolismo , Periodontite/patologia , Fagocitose , Fenótipo , Fatores de Tempo
8.
Environ Toxicol Pharmacol ; 88: 103755, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34662732

RESUMO

Periodontitis is a most prevalent and infectious multifactorial inflammatory disease and is characterized by the progressive destruction of the tooth-supporting tissues. Porphyromonas gingivalis, a Gram­negative oral anaerobe, mainly causes periodontitis and it is one of the most important risk factors responsible for aggravation of existing systemic diseases. Several experimental and clinical studies have shown the positive association between periodontitis and different forms of liver disease. Periodontal diseases increase the prevalence of non-alcoholic fatty liver diseases and cirrhosis. Infected periodontium and pathogens in the periodontal microenvironments release pathogen-associated molecular patterns such as peptidoglycan, lipopolysaccharides, gingipain, fimbria, bacterial DNA, etc, and damage-associated molecular patterns such as interleukins-1α, ß, - 8, and galectin-3, etc. These virulence factors and cytokines enter the bloodstream, disseminate into the whole body, and induce a variety of systemic pathological effects, including liver diseases (steatosis and fibrosis). Maintaining oral hygiene by scaling and root planning significantly improves liver damage in patients with periodontitis. Dentists and physicians should have more awareness in understanding the bidirectional nature of the relationship between oral and systemic diseases. Importantly, periodontitis condition aggravates simple fatty liver into fibrotic disease and therefore, the aim of this review is to understand the possible link between periodontitis and liver diseases.


Assuntos
Infecções por Bacteroidaceae/complicações , Endotoxinas/efeitos adversos , Hepatopatias/etiologia , Periodontite/complicações , Porphyromonas gingivalis , Animais , Humanos , Estilo de Vida , Higiene Bucal
9.
J Med Food ; 24(11): 1186-1190, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34698557

RESUMO

Black tea is a popular beverage worldwide. Theaflavins (TFs), which are active functional components of black tea, are potentially valuable for preventing and/or treating the progression of periodontal diseases. Our previous pilot study showed that TF intake decreases the number of Porphyromonas gingivalis (P. gingivalis) bacteria in the saliva. In this study, we aimed to determine whether TF intake improves periodontal disease attributed to oral bacteria in a randomized, placebo-controlled, and double-blind study. A total of 56 healthy subjects without periodontal diseases were enrolled and assigned to the placebo and TF groups (n = 28). TF intake for 6 weeks did not significantly alter the clinical evaluation of subjects. There was no significant adverse effect among the subjects. The number of P. gingivalis and Fusobacterium nucleatum (F. nucleatum) bacteria, which was the primary endpoint in this study, was not impacted by TF intake. The change ratio of Prevotella intermedia was significantly decreased by TF intake (P = .043) when compared with the placebo group. Collectively, our findings suggest that TFs have beneficial effects on oral bacteria for the prevention of periodontal disease. The study protocol was registered in the University Hospital Medical Information Network (UMIN000020049).


Assuntos
Fusobacterium nucleatum , Porphyromonas gingivalis , Aggregatibacter actinomycetemcomitans , Biflavonoides , Catequina , Método Duplo-Cego , Humanos , Japão , Projetos Piloto
10.
Chin J Dent Res ; 24(3): 167-175, 2021 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-34491011

RESUMO

OBJECTIVE: To analyse the subgingival microbiota of Stage I/II periodontitis, gingivitis with different degrees of severity, and periodontal health in subjects in a Chinese young adult population. METHODS: Subgingival plaque samples were collected from 15 Stage I/II periodontitis patients, 38 gingivitis patients and 15 periodontally healthy individuals, all aged from 18 to 21 years. Gingivitis patients were divided into two subgroups according to the Bleeding Index (BI) of their sampled teeth: gingivitis with above median BI (G-HBI) and below median BI (G-LBI). The subgingival plaque samples were collected from teeth 16, 26, 36, 46, 11 and 31 according to FDI notation. The V3-V4 region of the 16S rRNA gene of all the samples was sequenced and analysed. RESULTS: The Stage I/II periodontitis, gingivitis and periodontal health groups showed distinct subgingival microbiota profiles. When the gingivitis patients were stratified into two subgroups, the community structure of G-HBI showed no significant difference from early-stage periodontitis, but differed from G-LBI and the healthy group. Most periodontitis-related taxa were most abundant in Stage I/II periodontitis, followed by G-HBI, G-LBI and the periodontally healthy group. Porphyromonas gingivalis, Filifactor alocis, Tannerella forsythia, Saccharibacteria TM7 G-5 356, Lachnospiraceae G-8 500, Peptostreptococcaceae spp. and Syntrophomonadaceae VIIIG-1 435 were associated with Stage I/II periodontitis. Porphyromonas 275, Leptotrichia 417 and Saccharibacteria TM7 G-2 350 were associated with gingivitis. Porphyromonas gingivalis was significantly more abundant in G-HBI than in G-LBI. CONCLUSION: Within the limitations of this preliminary study, gingivitis and early-stage periodontitis were associated with an increased degree of dysbiosis in the subgingival microbiota in a Chinese young adult population.


Assuntos
Gengivite , Periodontite , China , Clostridiales , Nível de Saúde , Humanos , Porphyromonas gingivalis/genética , RNA Ribossômico 16S/genética
11.
J Biotechnol ; 340: 22-29, 2021 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-34478774

RESUMO

Porphyromonas gingivalis (P. gingivalis) is regarded as a keystone pathogen in destructive periodontal diseases. It expresses a variety of virulence factors, amongst them fimbriae that are involved in colonization, invasion, establishment and persistence of the bacteria inside the host cells. The fimbriae also were demonstrated to affect the host immune-response mechanisms. The major fimbriae are able to bind specifically to different host cells, amongst them peripheral blood monocytes. The interaction of these cells with fimbriae induces release of cytokines such as interleukin-1 (IL-1), IL-6, and tumor necrosis factor-α (TNF-α). The aim of this study was to generate recombinant major FimA protein from P. gingivalis W83 fimbriae and to prove its biological activity. FimA of P. gingivalis W83 was amplified from chromosomal DNA, cloned in a vector and transferred into Listeria innocua. (L. innocua).The expressed protein was harvested and purified using FPLC via a His trap HP column. The identity and purity was demonstrated by gel-electrophoresis and mass-spectrometry. The biological activity was assessed by stimulation of human oral epithelial cells and peripheral blood monocytes with the protein and afterwards cytokines in the supernatants were quantified by enzyme linked immunosorbent assay (ELISA) and cytometric bead array. Recombinant FimA could successfully be generated and purified. Gel-electrophoresis and mass-spectrometry confirmed that the detected sequences are identical with FimA. Stimulation of human monocytes induced the release of high concentrations of IL-1ß, IL-6, IL-10 and TNF-α by these cells. In conclusion, a recombinant FimA protein was established and its biological activity was proven. This protein may serve as a promising agent for further investigation of its role in periodontitis and possible new therapeutic approaches.


Assuntos
Listeria , Porphyromonas gingivalis , Proteínas de Fímbrias/genética , Fímbrias Bacterianas/genética , Humanos , Porphyromonas gingivalis/genética
12.
BMC Oral Health ; 21(1): 426, 2021 09 04.
Artigo em Inglês | MEDLINE | ID: mdl-34481478

RESUMO

BACKGROUND: B cell activating factor (BAFF) is a member of the tumor necrosis factor (TNF) superfamily with immunomodulatory effects on both innate and adaptive immune responses. Periodontitis is an inflammatory disease characterized by periodontal soft tissue inflammation and the progressive loss of periodontal ligament and alveolar bone. Macrophages are closely related to periodontitis progression. However, the role of BAFF in periodontitis development and macrophage polarization and the underlying mechanism remain unknown. METHODS: In vivo, a ligation-induced mouse model of periodontitis for BAFF blockade was established to investigate the expression of inducible nitric oxide synthase (iNOS) through real-time PCR (RT-PCR) and immunohistochemistry. In addition, the level of TNF-α in the periodontium, the number of osteoclasts, and alveolar bone resorption were observed. In vitro, RAW 264.7 macrophage cells were treated with 100 ng/mL Porphyromonas gingivalis lipopolysaccharide (P. gingivalis LPS) in either the presence or absence of 50 nM small interfering RNA (siRNA) targeting BAFF, followed by further incubation for 24 h. These cells and supernatants were collected and stored for RT-PCR, enzyme-linked immunosorbent assay, western blotting and immunofluorescence microscopy. RESULTS: In vivo, BAFF blockade decreased the levels of TNF-α in the periodontium in a ligature-induced mouse periodontitis model. Reduced osteoclast formation and lower alveolar bone loss were also observed. In addition, BAFF blockade was related to the expression of polarization signature molecules in macrophages. In vitro, BAFF knockdown notably suppressed the production of TNF-α in RAW 264.7 cells stimulated by P. gingivalis LPS. Moreover, BAFF knockdown attenuated the polarization of RAW 264.7 cells into classically activated macrophages (M1), with reduced expression of iNOS. CONCLUSIONS: Based on our limited evidence, we showed BAFF blockade exhibits potent anti-inflammatory properties in mice experimental periodontitis in vivo and in P. gingivalis LPS-treated RAW 264.7 cells in vitro, and macrophage polarization may be responsible for this effect.


Assuntos
Fator Ativador de Células B , Periodontite , Animais , Lipopolissacarídeos , Macrófagos , Camundongos , Porphyromonas gingivalis , Células RAW 264.7
13.
Shanghai Kou Qiang Yi Xue ; 30(3): 225-231, 2021 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-34476435

RESUMO

PURPOSE: To explore the effects of eicosapentaenoic acid (EPA) on biological activity and inflammatory factor expression of human gingival fibroblasts (HGFs). METHODS: The effects of EPA on the activity, morphology and cell cycle of HGFs were observed by living and dead cell staining, immunofluorescence staining and flow cytometry, respectively. HGFs were stimulated by lipopolysaccharides (LPS) of Porphyromonas gingivalis (P. gingivalis) or heat inactivated P. gingivalis, after which the effects of EPA on mRNA and protein expression of IL-6, IL-8 and IL-1ß were observed by real-time PCR and ELISA, respectively. The gene and protein expression of heme oxygenase-1(HO-1) was also detected by real-time PCR and Western blotting, respectively. The data were analyzed with SPSS 22.0 software package. RESULTS: 200 µmol/L EPA inhibited cell activity of HGFs; 100 µmol/L EPA did not affect cell activity and morphology of HGFs, and had no significant effect on cell cycle (P>0.05). EPA significantly downregulated gene expression of IL-6 and IL-1ß, and protein expression of IL-6 stimulated by P. gingivalis LPS and heat-killed P.gingivalis(P<0.05), in a dose-dependent manner. EPA increased gene expression of HO-1 in a dose dependent manner(P<0.05), and upregulated HO-1 protein expression. CONCLUSIONS: EPA significantly inhibits the expression of inflammatory factors without affecting the biological activity of HGFs, which may be related to the induction of HO-1, suggesting the potential role of EPA in the prevention and treatment of periodontitis.


Assuntos
Ácido Eicosapentaenoico , Gengiva , Células Cultivadas , Ácido Eicosapentaenoico/farmacologia , Fibroblastos , Humanos , Lipopolissacarídeos , Porphyromonas gingivalis
14.
Int J Oral Sci ; 13(1): 28, 2021 09 03.
Artigo em Inglês | MEDLINE | ID: mdl-34475379

RESUMO

Porphyromonas gingivalis (P. gingivalis), a key pathogen in periodontitis, has been shown to accelerate the progression of atherosclerosis (AS). However, the definite mechanisms remain elusive. Emerging evidence supports an association between mitochondrial dysfunction and AS. In our study, the impact of P. gingivalis on mitochondrial dysfunction and the potential mechanism were investigated. The mitochondrial morphology of EA.hy926 cells infected with P. gingivalis was assessed by transmission electron microscopy, mitochondrial staining, and quantitative analysis of the mitochondrial network. Fluorescence staining and flow cytometry analysis were performed to determine mitochondrial reactive oxygen species (mtROS) and mitochondrial membrane potential (MMP) levels. Cellular ATP production was examined by a luminescence assay kit. The expression of key fusion and fission proteins was evaluated by western blot and immunofluorescence. Mdivi-1, a specific Drp1 inhibitor, was used to elucidate the role of Drp1 in mitochondrial dysfunction. Our findings showed that P. gingivalis infection induced mitochondrial fragmentation, increased the mtROS levels, and decreased the MMP and ATP concentration in vascular endothelial cells. We observed upregulation of Drp1 (Ser616) phosphorylation and translocation of Drp1 to mitochondria. Mdivi-1 blocked the mitochondrial fragmentation and dysfunction induced by P. gingivalis. Collectively, these results revealed that P. gingivalis infection promoted mitochondrial fragmentation and dysfunction, which was dependent on Drp1. Mitochondrial dysfunction may represent the mechanism by which P. gingivalis exacerbates atherosclerotic lesions.


Assuntos
Dinâmica Mitocondrial , Porphyromonas gingivalis , Células Endoteliais , Mitocôndrias
15.
Biomater Sci ; 9(19): 6574-6583, 2021 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-34582534

RESUMO

Porphyromonas gingivalis, the pathogen of periodontal disease, is thought to be involved in various diseases throughout the body via gingival tissue blood capillaries. However, the dynamic analysis of the infection mechanism, particularly the deep invasion process of the gingival tissue, has not yet been elucidated because of the lack of both in vivo and in vitro models. In this study, we developed a vascularized three-dimensional (3D) gingival model with an epithelial barrier expressing cell-cell junctions using collagen microfibers (CMFs) to enable the dynamic analysis of the P. gingivalis invasion process. Lipid raft disruption experiments in the gingival epithelial cell layer demonstrated that P. gingivalis migrates into the deeper epithelium via the intercellular pathway rather than intracellular routes. P. gingivalis was shown to invade the 3D gingival model, being found inside blood capillaries during two days of culture. Notably, the number of bacteria had increased greatly at least two days later, whereas the mutant P. gingivalis lacking the cysteine proteases, gingipains, showed a significantly lower number of survivors. The secretion of interleukin-6 (IL-6) from the gingival tissue decreased during the two days of infection with the wild type P. gingivalis, but the opposite was found for the mutant suggesting that P. gingivalis infection disturbs IL-6 secretion at an early stage. By allowing the dynamic observation of the P. gingivalis invasion from the epithelial cell layer into the blood capillaries for the first time, this model will be a powerful tool for the development of novel therapeutics against periodontal infection related diseases.


Assuntos
Capilares , Porphyromonas gingivalis , Células Cultivadas , Células Epiteliais , Gengiva , Humanos
16.
Molecules ; 26(18)2021 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-34577061

RESUMO

Due to the limitations of traditional periodontal therapies, and reported cold atmospheric plasma anti-inflammatory/antimicrobial activities, plasma could be an adjuvant therapy to periodontitis. Porphyromonas gingivalis was grown in blood agar. Standardized suspensions were plated on blood agar and plasma-treated for planktonic growth. For biofilm, dual-species Streptococcus gordonii + P. gingivalis biofilm grew for 48 h and then was plasma-treated. XTT assay and CFU counting were performed. Cytotoxicity was accessed immediately or after 24 h. Plasma was applied for 1, 3, 5 or 7 min. In vivo: Thirty C57BI/6 mice were subject to experimental periodontitis for 11 days. Immediately after ligature removal, animals were plasma-treated for 5 min once-Group P1 (n = 10); twice (Day 11 and 13)-Group P2 (n = 10); or not treated-Group S (n = 10). Mice were euthanized on day 15. Histological and microtomography analyses were performed. Significance level was 5%. Halo diameter increased proportionally to time of exposure contrary to CFU/mL counting. Mean/SD of fibroblasts viability did not vary among the groups. Plasma was able to inhibit P. gingivalis in planktonic culture and biofilm in a cell-safe manner. Moreover, plasma treatment in vivo, for 5 min, tends to improve periodontal tissue recovery, proportionally to the number of plasma applications.


Assuntos
Periodontite/tratamento farmacológico , Gases em Plasma/uso terapêutico , Animais , Linhagem Celular , Quimioterapia Adjuvante/métodos , Chlorocebus aethiops , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Gases em Plasma/toxicidade , Porphyromonas gingivalis/efeitos dos fármacos , Streptococcus gordonii/efeitos dos fármacos , Células Vero
17.
Molecules ; 26(18)2021 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-34577123

RESUMO

Porphyromonas gingivalis has been identified as one of the major periodontal pathogens. Activity-directed fractionation and purification processes were employed to identify bioactive compounds from bitter melon leaf. Ethanolic extract of bitter melon leaf was separated into five subfractions by open column chromatography. Subfraction-5-3 significantly inhibited P. gingivalis-induced interleukin (IL)-8 and IL-6 productions in human monocytic THP-1 cells and then was subjected to separation and purification by using different chromatographic methods. Consequently, 5ß,19-epoxycucurbita-6,23(E),25(26)-triene-3ß,19(R)-diol (charantadiol A) was identified and isolated from the subfraction-5-3. Charantadiol A effectively reduced P. gingivalis-induced IL-6 and IL-8 productions and triggered receptors expressed on myeloid cells (TREM)-1 mRNA level of THP-1 cells. In a separate study, charantadiol A significantly suppressed P. gingivalis-stimulated IL-6 and tumor necrosis factor-α mRNA levels in gingival tissues of mice, confirming the inhibitory effect against P. gingivalis-induced periodontal inflammation. Thus, charantadiol A is a potential anti-inflammatory agent for modulating P. gingivalis-induced inflammation.


Assuntos
Monócitos , Porphyromonas gingivalis , Animais , Anti-Inflamatórios/farmacologia , Temperatura Alta , Camundongos , Momordica charantia , Periodontite
18.
Mol Oral Microbiol ; 36(6): 316-326, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34569151

RESUMO

The main etiological agent of periodontitis is the anaerobic bacterium Porphyromonas gingivalis. Virulence of this pathogen is controlled by various mechanisms and executed by major virulence factors including the gingipain proteases, peptidylarginine deiminase (PPAD), and RagB, an outer membrane macromolecular transport component. Although the structures and functions of these proteins are well characterized, little is known about their posttranslational maturation. Here, we determined the phosphoproteome of P. gingivalis in which phosphorylated tyrosine residues constitute over 80% of all phosphoresidues. Multiple phosphotyrosines were found in gingipains, PPAD, and RagB. Although mutation of phosphorylated residues in PPAD and RagB had no effect on secretion or activity, site-directed mutagenesis showed that phosphorylation in hemagglutinin/adhesin domains of RgpA and Kgp, and in the catalytic domain of RgpB, had a strong influence on secretion, processing, and enzymatic activity. Moreover, preventing phosphorylation of one gingipain influenced the others, suggesting multiple phosphorylation-dependent pathways of gingipain maturation in P. gingivalis. Various candidate kinases including Ptk1 BY kinase and ubiquitous bacterial kinase 1 (UbK1) may be involved, but their contribution to gingipain processing and activation remains to be confirmed.


Assuntos
Porphyromonas gingivalis , Fatores de Virulência , Adesinas Bacterianas/genética , Adesinas Bacterianas/metabolismo , Composição de Bases , Cisteína Endopeptidases/genética , Cisteína Endopeptidases/metabolismo , Hemaglutininas/genética , Fosforilação , Filogenia , Porphyromonas gingivalis/genética , Porphyromonas gingivalis/metabolismo , RNA Ribossômico 16S , Análise de Sequência de DNA , Fatores de Virulência/genética
19.
Clin Oral Investig ; 25(12): 6533-6546, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34495401

RESUMO

OBJECTIVE: This systematic review assesses the prevalence of microbial complexes in endodontic-periodontal lesion. MATERIALS AND METHODS: Nine databases were searched through August 2020. Experts were consulted to indicate additional studies. Studies were blindly selected by two reviewers based on pre-defined eligibility criteria. Studies that evaluated the prevalence of microbial orange and red complexes among patients with endodontic-periodontal lesion were considered eligible. Risk of bias was assessed using the Joanna Briggs Institute Critical Appraisal Checklist for Studies Reporting Prevalence Data. RESULTS: From 572 articles found on all databases, 11 clinical studies were finally included. The following microorganisms were investigated: P. gingivalis, T. forsythia, T. denticola, F. nucleatum, F. periodonticum, P. micra, P. intermedia, P. nigrescens, S. constellatus, C. gracilis, C. rectus, C. showae and E. nodatum. Considering the orange complex, P. micra, E. nodatum and S. constellatus were prevalent in both root canal and periodontal pockets. P. gingivalis and T. forsythia belonging to the red complex were prevalent only in periodontal pockets. The red complex microorganisms were not found very frequently in root canal. CONCLUSIONS: There is a similarity between the microbiome of root canal and periodontal pockets, with prevalence of the three microorganisms of the orange complex. Two microorganisms from the red complex were prevalent only in periodontal pockets. CLINICAL RELEVANCE: The prevalence of specific microorganisms in endodontic-periodontal lesion is important to understand the microbiological profile of the patients involved and to correlate it with possible clinical and repair conditions of this pathology.


Assuntos
Citrus sinensis , Microbiota , Humanos , Bolsa Periodontal , Porphyromonas gingivalis , Prevalência , Tratamento do Canal Radicular
20.
Artigo em Inglês | MEDLINE | ID: mdl-34574484

RESUMO

A particular role for Porphyromonas gingivalis (Pg) and Aggregatibacter actinomycetemcomitans (Aa) has been suggested in periodontitis and rheumatoid arthritis (RA), as these bacteria could initiate the formation of rheumatoid factor (RF) and anticitrullinated protein autoantibodies (ACPA). We assessed whether serum antibodies against Pg and Aa in RA patients and non-RA controls reflect the subgingival presence of Pg and Aa, and evaluated the relationship of these antibodies to the severity of periodontal inflammation and RA-specific serum autoantibodies. In 70 Indonesian RA patients and 70 non-RA controls, the subgingival presence of Pg and Aa was assessed by bacterial 16S rRNA gene sequencing, and serum IgG levels specific for Pg and Aa were determined. In parallel, serum levels of ACPA (ACPA:IgG,IgA) and RF (RF:IgM,IgA) were measured. The extent of periodontal inflammation was assessed by the periodontal inflamed surface area. In both RA patients and the controls, the presence of subgingival Pg and Aa was comparable, anti-Pg and anti-Aa antibody levels were associated with the subgingival presence of Pg and Aa, and anti-Pg did not correlate with ACPA or RF levels. The subgingival Pg and Aa were not related to RA. No noteworthy correlation was detected between the antibodies against Pg and Aa, and RA-specific autoantibodies.


Assuntos
Artrite Reumatoide , Fator Reumatoide , Autoanticorpos , Humanos , Porphyromonas gingivalis , RNA Ribossômico 16S/genética
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