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1.
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
2.
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 , /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
3.
Pol J Microbiol ; 68(3): 377-382, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31880883

RESUMO

The aim of this study was to assess the periodontal status of cystic fibrosis (CF) adult patients and to evaluate whether there is a correlation between the bacterial population of the subgingival biofilm and the health status of the periodontal tissues in this group of adults. The study involved 22 cystic fibrosis adult patients. The periodontal condition was assessed using Plaque Index (PLI), Gingival Index (GI), and Probing Pocket Depth (PPD). The gingival sulcus samples were analyzed by the Real-Time PCR assay (RT-PCR). Majority of patients showed moderate or severe bacterial dental plaque accumulation, but none of them had clinical symptoms of periodontal diseases. RT-PCR showed the presence of periopathogens in 50% of patients. Red complex microorganisms were detected in 9.09%, orange complex in 27.27%, and green complex in 31.82% of the samples analyzed. In cystic fibrosis patients colonized by periopathogens, the periodontal markers were significantly higher in comparison to not colonized by periopathogens patients. Despite the widespread presence of bacterial dental deposits in the cystic fibrosis adult patients examined, none of them has clinical symptoms of periodontal disease; however, the presence of periodontal pathogens in subgingival biofilm may represent a possible risk factor of this disease in the future. An unsatisfactory level of oral hygiene in any patient with cystic fibrosis indicates a need to focus on standards of dental care for such patients.The aim of this study was to assess the periodontal status of cystic fibrosis (CF) adult patients and to evaluate whether there is a correlation between the bacterial population of the subgingival biofilm and the health status of the periodontal tissues in this group of adults. The study involved 22 cystic fibrosis adult patients. The periodontal condition was assessed using Plaque Index (PLI), Gingival Index (GI), and Probing Pocket Depth (PPD). The gingival sulcus samples were analyzed by the Real-Time PCR assay (RT-PCR). Majority of patients showed moderate or severe bacterial dental plaque accumulation, but none of them had clinical symptoms of periodontal diseases. RT-PCR showed the presence of periopathogens in 50% of patients. Red complex microorganisms were detected in 9.09%, orange complex in 27.27%, and green complex in 31.82% of the samples analyzed. In cystic fibrosis patients colonized by periopathogens, the periodontal markers were significantly higher in comparison to not colonized by periopathogens patients. Despite the widespread presence of bacterial dental deposits in the cystic fibrosis adult patients examined, none of them has clinical symptoms of periodontal disease; however, the presence of periodontal pathogens in subgingival biofilm may represent a possible risk factor of this disease in the future. An unsatisfactory level of oral hygiene in any patient with cystic fibrosis indicates a need to focus on standards of dental care for such patients.


Assuntos
Biofilmes , Fibrose Cística/microbiologia , Doenças Periodontais/microbiologia , Adulto , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Fenômenos Fisiológicos Bacterianos , Feminino , Gengiva/microbiologia , Humanos , Masculino , Pessoa de Meia-Idade , Índice Periodontal , Periodonto/microbiologia , Adulto Jovem
4.
Adv Exp Med Biol ; 1197: 55-67, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31732934

RESUMO

Gingival epithelium plays a pivotal role in protecting the underlying periodontium from the microbial colonization found in the gingival sulcus. Having an appropriate phenotype displayed by gingival epithelial cells is a critical host component required for protection against bacterial invasion into gingival tissues. In the present study, gingival epithelial homeostasis associated with the CXCL-8/IL-8 chemokine response was investigated in vitro to determine the mechanisms that gingival epithelial cells utilize for sensing gram-positive and gram-negative microorganisms. The findings of this study have demonstrated, by using Fusobacterium nucleatum, a heterogeneity of gingival epithelial cell response by Toll-like receptor (TLR) 2, a lipoprotein sensor. Notably, however, lipopolysaccharide (LPS), a major virulence factor of gram-negative bacteria, is not recognized by gingival epithelial cells unless the LPS is internalized into the cells. Activation of TLR4 in gingival epithelial cells occurs in the endosome, an intracellular event that requires a vesicular acidification to turn on TLR4 signaling, indicating their stringency for fine-tuning a local LPS response. This study has identified a unique LPS sensing mechanism of the oral epithelium to overcome a periodontal infection associated with LPS derived from gram-negative microbes that arises during dysbiosis.


Assuntos
Gengiva , Lipopolissacarídeos , Periodontite , Células Epiteliais/imunologia , Células Epiteliais/microbiologia , Gengiva/citologia , Gengiva/imunologia , Gengiva/microbiologia , Humanos , Interleucina-8/imunologia , Lipopolissacarídeos/metabolismo , Periodontite/imunologia , Periodontite/microbiologia
5.
Braz Oral Res ; 33(suppl 1): e064, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31576948

RESUMO

The aim was of this study was to determine the current weight of evidence for the existence of specific differences between the microbiota of healthy teeth and healthy implants, or of teeth with periodontitis and implants with peri-implantitis. A systematic review was conducted according to the PRISMA statement. The MEDLINE, EMBASE and Cochrane databases were searched up to February 2018 for studies comparing microbiological data of biofilm samples collected from healthy teeth and implants or from teeth with periodontitis and implants with peri-implantitis. The weight of evidence was defined in three categories (strong, moderate and mild/some), according to the difference in number of studies showing statistically significantly higher counts and/or proportions and/or abundance and/or prevalence of microorganisms in health or in disease. Of the 132 articles identified, 8 were included. A wide range of microorganisms were present in different conditions but no microorganisms showed strong, moderate or mild/some evidence for a specific association with either teeth or implants. The results of this systematic review indicated that there is insufficient evidence in the literature to support specific differences between microorganisms colonizing teeth and implants, either in health or in disease.


Assuntos
Implantes Dentários/microbiologia , Gengiva/microbiologia , Peri-Implantite/microbiologia , Periodontite/microbiologia , Bactérias/isolamento & purificação , Biofilmes/crescimento & desenvolvimento , Estudos de Casos e Controles , Placa Dentária/microbiologia , Humanos , Microbiota
6.
Nat Commun ; 10(1): 4496, 2019 10 03.
Artigo em Inglês | MEDLINE | ID: mdl-31582750

RESUMO

Solitary chemosensory cells (SCCs) are epithelial sentinels that utilize bitter Tas2r receptors and coupled taste transduction elements to detect pathogenic bacterial metabolites, triggering host defenses to control the infection. Here we report that SCCs are present in mouse gingival junctional epithelium, where they express several Tas2rs and the taste signaling components α-gustducin (Gnat3), TrpM5, and Plcß2. Gnat3-/- mice have altered commensal oral microbiota and accelerated naturally occurring alveolar bone loss. In ligature-induced periodontitis, knockout of taste signaling molecules or genetic absence of gingival SCCs (gSCCs) increases the bacterial load, reduces bacterial diversity, and renders the microbiota more pathogenic, leading to greater alveolar bone loss. Topical treatment with bitter denatonium to activate gSCCs upregulates the expression of antimicrobial peptides and ameliorates ligature-induced periodontitis in wild-type but not in Gnat3-/- mice. We conclude that gSCCs may provide a promising target for treating periodontitis by harnessing innate immunity to regulate the oral microbiome.


Assuntos
Células Quimiorreceptoras/imunologia , Gengiva/imunologia , Imunidade Inata , Microbiota/imunologia , Periodontite/imunologia , Animais , Células Quimiorreceptoras/metabolismo , Modelos Animais de Doenças , Feminino , Gengiva/citologia , Gengiva/microbiologia , Células HEK293 , Proteínas Heterotriméricas de Ligação ao GTP/genética , Proteínas Heterotriméricas de Ligação ao GTP/metabolismo , Humanos , Masculino , Camundongos , Camundongos Knockout , Mucosa Bucal/citologia , Mucosa Bucal/imunologia , Mucosa Bucal/metabolismo , Periodontite/microbiologia , Fosfolipase C beta/metabolismo , Receptores Acoplados a Proteínas-G/metabolismo , Transdução de Sinais/imunologia , Canais de Cátion TRPM/metabolismo
7.
Ann Saudi Med ; 39(4): 244-250, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31381369

RESUMO

BACKGROUND: The periodontal tissues are continuously exposed to specific bacterial components that have the ability to alter many local functions. Normal endogenous infections in healthy mouths cause disease when their numbers increase significantly. OBJECTIVE: Determine the percentage of different periodontal pathogenic bacteria and their association with periodontal status. DESIGN: Cross-sectional, analytical. SETTINGS: School children of both genders in Saudi Arabia. PATIENTS AND METHODS: Clinical examination consisted of measurement of the gingival and periodontal supporting tissue including attachment loss, probing pocket depth and furcation involvement following the National Health and Nutrition Examination Survey (NHANES) and taking samples of the subgingival bacterial flora. MAIN OUTCOME MEASURES: The percentage of periodontal pathogenic bacteria and its association with periodontal status in Saudi Arabia. SAMPLE SIZE: Bacterial samples were collected from 277 subjects. RESULTS: Aggregatibacter actinomycetemcomitans was present in 21.7% of the subjects, Porphyromonas gingivalis in 21.3%; Tannerella forsythia in 10.1%; Treponema denticola in 34.7% and Prevotella inter-media in 12.3%. The red complex bacteria were found in 2.9% of the subjects. CONCLUSIONS: The percentages of bacteria varied but only T denticola was significantly associated with periodontal breakdown. In addition, the presence of more than 2 of the 5 species tested were significantly associated with tissue damage. LIMITATIONS: Cannot be generalized to all of Saudi Arabia. Larger controlled studies are needed. CONFLICT OF INTEREST: None.


Assuntos
Bactérias/isolamento & purificação , Gengiva/microbiologia , Doenças Periodontais/microbiologia , Periodonto/microbiologia , Adolescente , Estudos Transversais , Feminino , Humanos , Masculino , Doenças Periodontais/epidemiologia , Arábia Saudita/epidemiologia , Treponema denticola/isolamento & purificação
8.
Eur J Clin Microbiol Infect Dis ; 38(11): 2005-2019, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31372904

RESUMO

Human oral cavity (mouth) hosts a complex microbiome consisting of bacteria, archaea, protozoa, fungi and viruses. These bacteria are responsible for two common diseases of the human mouth including periodontal (gum) and dental caries (tooth decay). Dental caries is caused by plaques, which are a community of microorganisms in biofilm format. Genetic and peripheral factors lead to variations in the oral microbiome. It has known that, in commensalism and coexistence between microorganisms and the host, homeostasis in the oral microbiome is preserved. Nonetheless, under some conditions, a parasitic relationship dominates the existing situation and the rise of cariogenic microorganisms results in dental caries. Utilizing advanced molecular biology techniques, new cariogenic microorganisms species have been discovered. The oral microbiome of each person is quite distinct. Consequently, commonly taken measures for disease prevention cannot be exactly the same for other individuals. The chance for developing tooth decay in individuals is dependent on factors such as immune system and oral microbiome which itself is affected by the environmental and genetic determinants. Early detection of dental caries, assessment of risk factors and designing personalized measure let dentists control the disease and obtain desired results. It is necessary for a dentist to consider dental caries as a result of a biological process to be targeted than treating the consequences of decay cavities. In this research, we critically review the literature and discuss the role of microbial biofilms in dental caries.


Assuntos
Biofilmes/crescimento & desenvolvimento , Cárie Dentária/microbiologia , Microbiota/fisiologia , Boca/microbiologia , Bactérias/isolamento & purificação , Bactérias/patogenicidade , Cárie Dentária/genética , Cárie Dentária/prevenção & controle , Doenças da Polpa Dentária/genética , Doenças da Polpa Dentária/microbiologia , Doenças da Polpa Dentária/prevenção & controle , Gengiva/microbiologia , Gengiva/fisiologia , Humanos , Doenças Periodontais/genética , Doenças Periodontais/microbiologia , Doenças Periodontais/prevenção & controle , Saliva/química
9.
Pesqui. vet. bras ; 39(7): 454-459, July 2019. tab
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1040710

RESUMO

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


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


Assuntos
Animais , Doenças Periodontais/veterinária , Periodontite/veterinária , Ovinos , Gengiva/microbiologia , Bactérias Anaeróbias , Reação em Cadeia da Polimerase/veterinária , Microbiota
10.
Genome Res ; 29(8): 1352-1362, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31160374

RESUMO

Predicting biosynthetic gene clusters (BGCs) is critically important for discovery of antibiotics and other natural products. While BGC prediction from complete genomes is a well-studied problem, predicting BGCs in fragmented genomic assemblies remains challenging. The existing BGC prediction tools often assume that each BGC is encoded within a single contig in the genome assembly, a condition that is violated for most sequenced microbial genomes where BGCs are often scattered through several contigs, making it difficult to reconstruct them. The situation is even more severe in shotgun metagenomics, where the contigs are often short, and the existing tools fail to predict a large fraction of long BGCs. While it is difficult to assemble BGCs in a single contig, the structure of the genome assembly graph often provides clues on how to combine multiple contigs into segments encoding long BGCs. We describe biosyntheticSPAdes, a tool for predicting BGCs in assembly graphs and demonstrate that it greatly improves the reconstruction of BGCs from genomic and metagenomics data sets.


Assuntos
Genes Bacterianos , Metagenoma , Metagenômica/métodos , Família Multigênica , Software , Mapeamento de Sequências Contíguas , Conjuntos de Dados como Assunto , Placa Dentária/microbiologia , Gengiva/microbiologia , Humanos , Internet , Mucosa Bucal/microbiologia , Faringe/microbiologia , Biossíntese de Proteínas , Língua/microbiologia
11.
Microbiol Immunol ; 63(8): 293-302, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31209914

RESUMO

Antimicrobial peptides play important roles in the innate immune system of various organisms, and they may also be considered to prevent the organisms from infections. In particular, ß-defensins, mainly produced in epithelial cells, are recognized as one of the major antimicrobial peptides in mammals, including humans. In this study, we showed that Lactobacillus helveticus SBT2171 (LH2171), one of the several species of lactic acid bacteria, upregulates the production of ß-defensins in oral epithelial cells in vitro. Moreover, LH2171 reduced the increase of proinflammatory cytokine expression, induced by Porphyromonas gingivalis stimulation, in gingival epithelial cells. These data suggested that LH2171 suppresses P. gingivalis-induced inflammation by upregulating the expression of ß-defensins in gingival epithelial cells. We subsequently investigated the effects of LH2171 in vivo and revealed that ß-defensin expression was increased in the oral cavities of LH2171-fed mice. Furthermore, LH2171 decreased alveolar bone loss, gingival inflammation, and amounts of P. gingivalis-specific 16S ribosomal RNA in the gingiva of P. gingivalis-inoculated mice. Taken together, our results showed that LH2171 upregulates the expression of ß-defensins in oral cavity, thereby decreasing the number of P. gingivalis consequently ameliorating the experimental periodontal disease.


Assuntos
Células Epiteliais/metabolismo , Lactobacillus helveticus/fisiologia , Doenças Periodontais/prevenção & controle , Porphyromonas gingivalis/efeitos dos fármacos , Regulação para Cima , beta-Defensinas/metabolismo , beta-Defensinas/farmacologia , Perda do Osso Alveolar/prevenção & controle , Animais , Citocinas/metabolismo , Modelos Animais de Doenças , Células Epiteliais/microbiologia , Feminino , Gengiva/metabolismo , Gengiva/microbiologia , Camundongos , Camundongos Endogâmicos BALB C , Doenças Periodontais/microbiologia , Porphyromonas gingivalis/genética , RNA Mensageiro/metabolismo , RNA Ribossômico 16S/genética
12.
Photobiomodul Photomed Laser Surg ; 37(5): 288-297, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31084561

RESUMO

Objective: The aim of this study was to examine effects of recently developed ultraviolet light-emitting diodes (UV LEDs) wavelengths on in vitro growth and gene expression of cultural periodontopathic bacteria, and on viability of experimental gingival fibroblasts. Materials and methods: Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, and Streptococcus oralis were irradiated by UV LEDs (265, 285, 310, 365, and 448 nm) at 600 mJ/cm2 and grown anaerobically in vitro. The colony forming units were counted after 1 week. Cell morphology was observed using a scanning electron microscope (SEM). Quantitative real-time polymerase chain reaction was performed to investigate gene expression changes by 310 nm irradiation. Viability of the irradiated human gingival fibroblasts was evaluated using WST-8 assay. Results: Both 265 and 285 nm resulted in the complete death of bacteria and fibroblasts, whereas 310 nm caused partial killing and suppression of bacterial growth and much less damage to the fibroblasts in vitro. Both 365 and 448 nm resulted in no significant change. SEM showed that P. gingivalis cells gradually degraded from day 2 or 3 and were severely destructed on day 5 for 265, 285, and 310 nm. The 310 nm irradiation transiently suppressed the transcripts of SOS response- and cell division-relative genes. Conclusions: Both 265 and 285 nm may induce powerful bactericidal effects and severe fibroblast phototoxicity, and 310 nm may induce partial killing or growth suppression of bacterial cells with much less fibroblast phototoxicity. UV lights may have potential for bacterial suppression, with situations dependent on wavelength, in periodontal and peri-implant therapy.


Assuntos
Aggregatibacter actinomycetemcomitans/efeitos da radiação , Fusobacterium nucleatum/efeitos da radiação , Porphyromonas gingivalis/efeitos da radiação , Prevotella intermedia/efeitos da radiação , Streptococcus oralis/efeitos da radiação , Terapia Ultravioleta , Técnicas de Cultura de Células , Fibroblastos/efeitos da radiação , Gengiva/microbiologia , Gengiva/patologia , Gengiva/efeitos da radiação , Humanos , Células-Tronco
13.
Biomed Res Int ; 2019: 8456342, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30956987

RESUMO

Dental implantology allows replacement of failing teeth providing the patient with a general improvement of health. Unfortunately not all reconstructions succeed, as a consequence of the development of infections of bacterial origin on the implant surface. Surface topography is known to modulate a differential response to bacterial and mammalian cells but topographical measurements are often limited to vertical parameters. In this work we have extended the topographical measurements also to lateral and hybrid parameters of the five most representative implant and prosthetic component surfaces and correlated the results with bacterial and mammalian cell adhesion and proliferation outcomes. Primary human oral gingival fibroblast (gum cells) and the bacterial strains: Streptococcus mutans, Streptococcus sanguinis and Aggregatibacter actinomycetemcomitans, implicated in infectious processes in the oral/implant environment were employed in the presence or absence of human saliva. The results confirm that even though not all the measured surface is available for bacteria to adhere, the overall race for the surface between cells and bacteria is more favourable to the smoother surfaces (nitrided, as machined or lightly acid etched) than to the rougher ones (strong acid etched or sandblasted/acid etched).


Assuntos
Aderência Bacteriana , Implantes Dentários/microbiologia , Fibroblastos , Gengiva , Bactérias Gram-Positivas/crescimento & desenvolvimento , Mucosa Bucal , Adesão Celular , Fibroblastos/metabolismo , Fibroblastos/microbiologia , Fibroblastos/patologia , Gengiva/metabolismo , Gengiva/microbiologia , Gengiva/patologia , Bactérias Gram-Positivas/classificação , Bactérias Gram-Positivas/isolamento & purificação , Humanos , Mucosa Bucal/metabolismo , Mucosa Bucal/microbiologia , Mucosa Bucal/patologia , Propriedades de Superfície
14.
PLoS One ; 14(3): e0213309, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30870452

RESUMO

Porphyromonas gulae, an animal periodontal pathogen, possess fimbriae classified into three genotypes (A-C) based on the diversity of fimA genes encoding FimA. Accumulating evidence suggests that P. gulae strains with type C fimbriae are more virulent as compared to those with other types. The ability of these organisms to adhere to and invade gingival epithelial cells has yet to be examined. P. gulae showed the greatest levels of adhesion and invasion at a multiplicity of infection of 100 for 90 min. P. gulae type C and some type B strains invaded gingival epithelial cells at significantly greater levels than the other strains, at the same level of efficiency as P. gingivalis with type II fimbriae. Adhesion and invasion of gingival epithelial cells by P. gulae were inhibited by cytochalasin D and sodium azide, indicating the requirements of actin polymerization and energy metabolism for those activities. Invasion within gingival epithelial cells was blocked by staurosporine, whereas those inhibitors showed little effects on adhesion, while nocodazole and cycloheximide had negligible effects on either adhesion or invasion. P. gulae proteases were found to be essential for adhesion and invasion of gingival epithelial cells, while its DNA and RNA, and protein synthesis were unnecessary for those activities. Additionally, α5ß1 integrin antibodies significantly inhibited adhesion and invasion by P. gulae. This is the first report to characterize P. gulae adhesion and invasion of human gingival epithelial cells.


Assuntos
Aderência Bacteriana , Infecções por Bacteroidaceae/microbiologia , Células Epiteliais/microbiologia , Fímbrias Bacterianas/microbiologia , Gengiva/microbiologia , Porphyromonas/isolamento & purificação , Células Epiteliais/citologia , Gengiva/citologia , Humanos , Integrina alfa5beta1/metabolismo
15.
Mil Med ; 184(Suppl 1): 97-101, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30901402

RESUMO

OBJECTIVES: Sodium lauryl sulfate (SLS) is a surfactant used to decrease the surface tension of water. Most commercially available dentifrices contain 0.5-2.0% SLS. This study investigated the potential effect of SLS on oral wound healing using primary human gingival fibroblasts (HGFs). METHODS: HGFs cells were grown in12-well culture plates in DMEM medium. A 3 mm wound was created on confluent HGFs. The cells were challenged with 0 (the control group), 0.01, 0.02, 0.03, 0.04, or 0.05% SLS-containing media once daily for 2 minutes. The cells were stained on day 0, 2, 4, 6 and 8. The percent of wound fill area was measured. RESULTS: On day 2, 4, 6, and 8, the wound fill of the control group (0% SLS) was 15, 35, 67 and 98%, respectively; at 0.01% SLS, it was 10, 20, 65 and 84%; at 0.02%, it was 7, 10, 15 and 25%; at 0.03% SLS, it was only 5% and 8% on day 2 and 4. CONCLUSION: Our results showed a dose- and time-dependent inhibition on HGFs wound fill by SLS; however, future in vivo studies are needed to validate if our in vitro findings using SLS-free dentifrices to avoid the potential delay of wound healing.


Assuntos
Fibroblastos/efeitos dos fármacos , Gengiva/efeitos dos fármacos , Dodecilsulfato de Sódio/farmacologia , Cicatrização/efeitos dos fármacos , Relação Dose-Resposta a Droga , Fibroblastos/microbiologia , Gengiva/microbiologia , Humanos , Dodecilsulfato de Sódio/uso terapêutico , Tensoativos/farmacologia , Tensoativos/uso terapêutico , Fatores de Tempo
16.
Artigo em Inglês | MEDLINE | ID: mdl-30915280

RESUMO

Periodontal microorganisms not only colonize subgingival pockets, but also are detected on various mucous membranes in patients with periodontitis. The object of this pilot study was, using the next-generation sequencing of 16S RNA gene, to characterize the microbiota in two oral habitats (buccal mucosas and subgingival pockets) in patients with different forms of periodontitis. Thirty-two buccal swab samples and 113 subgingival samples were obtained from eleven subjects with chronic periodontitis (ChP), twelve subjects with aggressive periodontitis (AgP), and nine periodontally healthy individuals (HP). Using Miseq Sequencing of 16S rRNA gene, we found that the subgingival and buccal mucosa microbiome of ChP and AgP patients both differed from HP. Meanwhile, Veillonella, Treponema, Filifactor, Fretibacterium, Peptostreptococcaceae_[XI][G-6], Peptostreptococcaceae_[XI][G-5], Bacteroidetes_[G-5], Bacteroidetes_[G-3], Peptostreptococcaceae_[XI][G-4], Peptostreptococcaceae_[XI][G-2] significantly increased both in buccal and subgingival plaque samples in periodontitis subjects (ChP and AgP) compared with HP. Moreover, the results based on the Unweighted UniFrac distance showed that buccal and subgingival plaque samples from the same individuals show higher community divergence than same habitats from different subject samples. This study demonstrated that the microbiome of buccal mucosa can be influenced by periodontitis. However, subgingival and buccal mucosa microbiome seem to be characterized by species-specific colonization patterns. This pilot study provides a glimpse at the changes of subgingival and buccal mucosa associated with periodontitis from a holistic view. Further studies should be taken to illuminate the interplay between these detected changes and periodontitis development.


Assuntos
Periodontite Agressiva/microbiologia , Gengiva/microbiologia , Microbiota , Mucosa Bucal/microbiologia , Adulto , Doença Crônica , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Feminino , Humanos , Masculino , Metagenômica , Filogenia , Projetos Piloto , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Adulto Jovem
17.
Methods Mol Biol ; 1922: 525-548, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30838598

RESUMO

Early childhood caries (ECC) is a biofilm-mediated disease. Social, environmental, and behavioral determinants as well as innate susceptibility are major influences on its incidence; however, from a pathogenetic standpoint, the disease is defined and driven by oral dysbiosis. In other words, the disease occurs when the natural equilibrium between the host and its oral microbiome shifts toward states that promote demineralization at the biofilm-tooth surface interface. Thus, a comprehensive understanding of dental caries as a disease requires the characterization of both the composition and the function or metabolic activity of the supragingival biofilm according to well-defined clinical statuses. However, taxonomic and functional information of the supragingival biofilm is rarely available in clinical cohorts, and its collection presents unique challenges among very young children. This paper presents a protocol and pipelines available for the conduct of supragingival biofilm microbiome studies among children in the primary dentition, that has been designed in the context of a large-scale population-based genetic epidemiologic study of ECC. The protocol is being developed for the collection of two supragingival biofilm samples from the maxillary primary dentition, enabling downstream taxonomic (e.g., metagenomics) and functional (e.g., transcriptomics and metabolomics) analyses. The protocol is being implemented in the assembly of a pediatric precision medicine cohort comprising over 6000 participants to date, contributing social, environmental, behavioral, clinical, and biological data informing ECC and other oral health outcomes.


Assuntos
Bactérias/genética , Biofilmes , Cárie Dentária/microbiologia , Metabolômica/métodos , Metagenômica/métodos , Dente Decíduo/microbiologia , Bactérias/isolamento & purificação , Bactérias/metabolismo , Pré-Escolar , DNA Bacteriano/genética , Cárie Dentária/etiologia , Perfilação da Expressão Gênica/métodos , Gengiva/microbiologia , Humanos , Microbiota , RNA Bacteriano/genética , Análise de Sequência de DNA/métodos , Análise de Sequência de RNA/métodos , Software , Manejo de Espécimes/métodos , Transcriptoma
18.
Arch Oral Biol ; 101: 92-99, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30909081

RESUMO

OBJECTIVE: The aim of the study was to profile the subgingival microbiome of Chinese adults with generalized aggressive periodontitis (GAgP) using human oral microbe identification microarray (HOMIM), and to compare the results with matched periodontal healthy controls. DESIGN: 15 subjects with GAgP and 15 age- and gender- matched periodontal healthy controls were included. Subgingival plaque samples were collected from the deepest pockets of patients with GAgP and matched sites in controls and then analyzed by 16S rRNA-based microarrays. Student's paired t-test was used to compare clinical parameters and mean number of bacterial taxa detected between the two groups. Fisher's exact probability test and Wilcoxon Rank Sum were used to compare bacterial species between all samples. A multiple linear regression model was used for correlations among age, gender and bacterial with clinical parameters. RESULTS: From a total sum of 379 strains tested, 171 bacterial strains were detected from subgingival plaques of the GAgP patients, more than the 157 strains detected in control group. Mean number of subgingival bacterial taxa detected in GAgP group was 68 (SD = 21.06) while in control group was 45 (SD = 21.60). 47 bacterial taxa were detected more frequently in GAgP group while 12 taxa were more prevalent in control group. The significantly more prevalent and abundant taxa of bacteria in GAgP group included Filifactor alocis, Desulfobulbus sp., Fretibacterium sp., Porphyromonas gingivalis, Tannerella forsythia, Porphyromon as endodontalis, Peptostreptococcaceae spp., Parvimonas micra, Eubacterium nodatum and Eubacterium saphenum. Meanwhile the more abundant taxa in control group were Streptococcus spp. and Pseudomonas aeruginosa. CONCLUSIONS: There are more taxa of bacteria in subgingival plaques of Chinese patients with GAgP than in healthy controls. F. alocis, Desulfobulbus sp., Fretibacterium sp., P. gingivalis and T. forsythia are strongly associated with GAgP. High-throughout microbiological results may help dentists have a better understanding of subgingival microbiome of GAgP.


Assuntos
Periodontite Agressiva/microbiologia , Gengiva/microbiologia , Microbiota , Adulto , Grupo com Ancestrais do Continente Asiático , Bactérias/classificação , Estudos de Casos e Controles , Humanos , RNA Ribossômico 16S/genética
19.
Microbiol Immunol ; 63(3-4): 100-110, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30817027

RESUMO

Aggregatibacter actinomycetemcomitans is considered to be associated with periodontitis. Leukotoxin (LtxA), which destroys leukocytes in humans, is one of this bacterium's major virulence factors. Amounts of neutrophil elastase (NE), which is normally localized in the cytoplasm of neutrophils, are reportedly increased in the saliva of patients with periodontitis. However, the mechanism by which NE is released from human neutrophils and the role of NE in periodontitis is unclear. In the present study, it was hypothesized that LtxA induces NE release from human neutrophils, which subsequently causes the breakdown of periodontal tissues. LtxA-treatment did not induce significant cytotoxicity against human gingival epithelial cells (HGECs) or human gingival fibroblasts (HGFs). However, it did induce significant cytotoxicity against human neutrophils, leading to NE release. Furthermore, NE and the supernatant from LtxA-treated human neutrophils induced detachment and death of HGECs and HGFs, these effects being inhibited by administration of an NE inhibitor, sivelestat. The present results suggest that LtxA mediates human neutrophil lysis and induces the subsequent release of NE, which eventually results in detachment and death of HGECs and HGFs. Thus, LtxA-induced release of NE could cause breakdown of periodontal tissue and thereby exacerbate periodontitis.


Assuntos
Aggregatibacter actinomycetemcomitans/metabolismo , Células Epiteliais/patologia , Exotoxinas/metabolismo , Fibroblastos/patologia , Gengiva/microbiologia , Elastase de Leucócito/metabolismo , Neutrófilos/patologia , Periodontite/microbiologia , Aggregatibacter actinomycetemcomitans/patogenicidade , Morte Celular/fisiologia , Linhagem Celular , Células Epiteliais/microbiologia , Fibroblastos/microbiologia , Gengiva/citologia , Glicina/análogos & derivados , Glicina/farmacologia , Humanos , Elastase de Leucócito/antagonistas & inibidores , Neutrófilos/microbiologia , Sulfonamidas/farmacologia , Fatores de Virulência/metabolismo
20.
J Periodontal Res ; 54(4): 405-412, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30734922

RESUMO

BACKGROUND AND OBJECTIVE: Metal-based dental restorations with a subgingival outline may enhance plaque accumulation and bacterial colonization. This study aimed to investigate whether metal-based restorations influence the composition of subgingival microbiome. MATERIAL AND METHODS: Per subject one site with a metal-based restoration and one contra-lateral site without a restoration were selected on basis of radiographic bone loss ≤2 mm, restoration outline at sulcus level/subgingivally, pocket depth ≤4 mm, and no root canal treatments. Subgingival samples were collected with sterile paper-points, and microbial profiles were obtained by 16S rRNA gene amplicon sequencing. Restorations were sampled with an Arkansas-stone and the metal composition was determined using energy-dispersive X-ray spectroscopy. RESULTS: A total of 22 sites from 11 subjects were included. No significant differences for the clinical parameters were found between the restored and unrestored sites. The average age of the restorations was 14.9 ± 7.1 years. Firmicutes was the most prevalent phylum at the restored sites (32% vs 20% of the reads of the unrestored sites, P = 0.016), and Actinobacteria at the unrestored sites (33% vs 18% of the reads of the restored sites, P = 0.01). Overall, sequences clustered into 573 operational taxonomic units (OTUs). Species richness of the restored sites was significantly higher than species richness of the unrestored sites (117 ± 32 and 96 ± 20 OTUs, respectively, P = 0.013). No associations between the metal composition and bacterial profiles were found. CONCLUSION: This study shows that metal-based restorations may enhance colonization of Firmicutes and the neighboring pocket may harbor more diverse microbial communities.


Assuntos
Actinobacteria/classificação , Materiais Dentários/química , Firmicutes/classificação , Gengiva/microbiologia , Metais/química , Microbiota , Adulto , Estudos Transversais , Placa Dentária/microbiologia , Restauração Dentária Permanente , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , RNA Ribossômico 16S/genética
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