Diferencia en la expresión del ligando de receptor activador para el factor nuclear K B, en tejido tumoral frente a la infección por diferentes patógenos periodontales / Difference in receptor activator of nuclear factor K B expression in tumoral tissue by different periodontal pathogens

La enfermedad periodontal es una de las principales causas de pérdida dentaria. Clínicamente, esta patología, mediada por la desregulación del sistema inmune producto de una disbiosis ocurrida en el surco gingival, inicia con la inflamación de la encía y evoluciona con el daño irreversible de los tejidos que rodean el diente. El hueso alveolar es uno de los tejidos afectados esta patología, esto debido a la activación de osteoclastos por la sobreexpresión de la proteína RANKL en el huésped. El propósito de este trabajo es determinar el nivel de sobreexpresión de RANKL, en un modelo de células tumorales U2OS, frente a la infección con Porphyromonas gingivalis y Prevotella intermedia. Para identificar el nivel de RANKL, se definieron cuatro grupos: Un grupo control, no tratado; Grupo PG, tratado con P. gingivalis; Grupo PI, tratado con P. Intermedia; y un grupo PG+PI, tratado con ambas bacterias. El nivel relativo de la proteína RANKL fue determinado en el sobrenadante y en los extractos celulares de manera independiente, mediante la técnica Western blot. En sobrenadantes, el grupo PG mostró mayores niveles de RANKL comparados con PI (p < 0,05). En extractos celulares los niveles fueron mayores en el grupo PG+PI (p < 0,05). El grupo PI mostró los niveles más bajos de RANKL. La infección polimicrobiana resulta en una mayor expresión de RANKL en células tumorales U2OS, mientras que frente a la infección P. gingivalis, se observó mayor cantidad de RANKL soluble.

SUMMARY: Periodontal disease is one of the main causes of tooth loss. Clinically, this pathology, mediated by the deregulation of the immune system due to a dysbiosis occurred in the gingival sulcus, begins with the inflammation of the gum and evolves with the irreversible damage of the tissues that surround the tooth. Alveolar bone is one of the most affected tissues by this disease, due to the activation of osteoclasts by the upregulation of RANKL in the host. The aim of this study is to determine the increase of RANKL, in a U2OS tumor cells model, inoculated with Porphyromonas gingivalis and Prevotella intermedia. To identify the level of RANKL, four groups were defined: A control group, not treated; PG group, treated with P.gingivalis; PI group, treated with P. intermedia; and a PG+PI group, treated with both bacteria. The relative level of RANKL was determined in the supernatant and cell extracts independently, using the Western blot technique. In supernatants, the PG group showed higher RANKL levels compared to PI (p < 0.05). In cell extracts the levels were higher in the PG+PI group (p < 0.05.). The PI group showed the lowest levels of RANKL.Polymicrobial infection results in a greater expression of of soluble RANKL was observed.

Association between Volatile Sulfur Compounds Prevotella intermedia and Matrix Metalloproteinase-8 Expression

Abstract Objective: To determine the correlation between levels of methyl mercaptan (CH3SH) hydrogen sulfide (H2S), the proportion of Prevotella intermedia (Pi), and matrix metalloproteinase-8 (MMP-8) gene expression levels in periodontitis patients accompanied by halitosis. Material and Methods: Samples were obtained from gingival crevicular fluid (GCF) in the deepest pocket and by swabbing in the tongue coating area in patients with periodontitis presenting with halitosis (n = 23) and healthy subjects as controls (n = 7). The values of CH3SH and H2S were obtained using Oral Chroma. The proportion of Pi and MMP-8 expression levels were evaluated using PCR-RT. All the result was statistically analyzed using SPSS software. Results: The levels of CH3SH and H2S in participants with PD ≥ 6 mm showed a robust negative correlation with the proportion of P. intermedia in GCF and tongue coating. No statistically significant association was detected between CH3SH and H2S levels and MMP-8 expression levels (p>0.05). Conclusion: There is no association between CH3SH and H2S levels, the proportion of P. intermedia, and MMP-8 expression in patients with periodontitis accompanied by halitosis (AU).

Prevalencia de Prevotella spp y Porphyromona spp en periodontitis crónica / Prevalence of Prevotella spp and Porphyromona spp in chronic periodontitis

Las enfermedades del periodonto tienen una etiopatogenia compleja y puede considerarse multifactorial. El factor etiológico esencial en la patología inflamatoria periodontal es la biopelícula dental y cuando el desequilibrio entre el huésped y los microorganismos cambia la complejidad de la flora. Ciertas bacterias como Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Prevotella loescheii, Fusobacterium nucleatum, Tannerrella forsythia, Campylobacter rectus, Eikenella corrodens y Treponema spp., han sido comúnmente relacionadas con la periodontitis crónica y son consideradas como indicadores de riesgo para la progresión de dicha enfermedad. El objetivo de este trabajo fue establecer la prevalencia de Prevotella spp y Porphyromona spp en los distintos estadios de periodontitis crónicas. Material y métodos: Se estudiaron 48 pacientes sistémicamente saludables con diagnóstico de periodontitis crónica. Se completó el consentimiento informado, se realizó historia clínica y examen periodontal. El estado periodontal se clasificó en distintos grados de severidad: leve, moderada y severa. Se tomaron muestras de dos sitios con mayor profundidad de sondaje con conos de papel absorbente estériles y se transportaron en un medio prerreducido. Para el aislamiento de Prevotella spp se utilizó agar Brucella más sangre ovina al 5%, hemina, vitamina K al que se agregaron vancomicina y kanamicina; Porphyromonas sp se aisló en el mismo medio con el agregado de bacitracina y colistina. Se sembraron 10 µl de muestra entera y las placas fueron incubadas en jarras de anaerobiosis por 5 a 7 días a 37ºC. Resultados: los distintos grados de periodontitis correspondieron a un 17% periodontits leve, 57% moderada y 26% severa. En el total de pacientes se determinó la presencia de Prevotella spp en el 54% de los casos y un 12,5% de Porphyromona spp. Conclusión: De los pacientes estudiados con periodontits crónica, un 52% correspondió al sexo masculino, un 57% de los casos correspondieron a periodontitis moderada. Se aisló Prevotella sp en todos los estadios de periodontitis crónica y Porphyromonas sp sólo en periodontitis severas (AU)

Periodontal diseases have a complex etiopathogenesis and can be considered multifactorial. The essential etiological factor in periodontal inflammatory pathology is the dental biofilm and when the imbalance between the host and the microorganisms changes the complexity of the flora. Certain bacteria such as Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Prevotella loescheii, Fusobacterium nucleatum, Tannerrella forsythia, Campylobacter rectus, Eikenella corrodens and Treponema spp., Have been commonly related to chronic periodontitis and are considered as risk indicators for the progression of said disease. The objective of this work was to establish the prevalence of Prevotella spp and Porphyromonas spp in the different stages of chronic periodontitis. Forty eight systemically healthy patients diagnosed with chronic periodontitis were studied. Informed consent was completed, a medical history and periodontal examination was carried out. The periodontal state was classified into different degrees of severity: mild, moderate and severe. Samples were taken from two sites with greater depth of probing with sterile absorbent paper cones and transported in a prereduced medium. For the isolation of Prevotella spp, Brucella agar plus 5% sheep blood, hemin, vitamin K to which vancomycin and kanamycin were added. For Porphyromonas spp, the same medium was used and bacitracin and colistin were added. 10 µl of the whole sample was seeded and the plates were incubated in anaerobic jars for 5 to 7 days at 37 ° C. Different degrees of periodontitis corresponded to 17% mild periodontitis, 57% moderate and 26% severe. In the total number of patients, the presence of Prevotella spp was determined in 54% of the cases and 12.5% of Porphyromona spp. Of the patients studied with chronic periodontitis, 52% corresponded to the male sex, 57% of the cases corresponded to moderate periodontitis. Prevotella spp was isolated in all stages of chronic periodontitis and Porphyromonas sp only in severe periodontitis (AU)

Influência de laminados cerâmicos cimentados sobre dentes naturais sem término cervical na microbiota do fluido gengival crevicular: estudo clínico, prospectivo e longitudinal usando PCR em tempo real / Influence of laminate veneers cemented over natural teeth without finish line on the microbiota of gingival crevicular fluid: clinical, prospective and longitudinal study using real-time PCR

Objetivo: quantificar bactérias colonizadoras do sulco gengival em paralelo à verificação do comportamento clínico periodontal e da adaptação marginal de laminados cerâmicos cimentados sobre dentes sem término cervical. Métodos: 73 laminados cerâmicos em dissilicato de lítio monolítico foram cimentados sobre dentes naturais sem término cervical e com a margem cervical posicionada cerca de 0,5mm no interior do sulco gengival. Para todos os dentes foi feita a coleta do fluido gengival crevicular (FGC) previamente à cimentação da restauração (baseline) e em 7, 180 e 365 dias após a cimentação para quantificação das bactérias S. mitis, P. intermedia e P. gingivalis através de PCR em tempo real. Clinicamente foram avaliados os parâmetros índice de placa visível (IPV), sangramento à sondagem (SS), profundidade de sondagem (PS), perda de inserção clínica (PIC), recessão gengival (RG) e a adaptação marginal da restauração em baseline, 7, 15, 30, 60, 180 e 365 dias. Foram obtidas réplicas da região cervical para análise da adaptação marginal por microscopia eletrônica de varredura (MEV). Resultados: Não houve diferença estatisticamente significante para o IPV, PS, SS em todos os períodos analisados (Anova, p>0,05). Não houve registro de PIC e RG. Houve diferença estatisticamente significante para S. mitis na comparação entre 180 e 365 dias (Dunn, p=0,03). Não foi detectado P. intermedia em nenhum dos sítios e não houve diferença estatisticamente significante para P. gingivalis em todos os tempos do estudo (Friedman, p>0,05). Todas as restaurações receberam o conceito alfa para a adaptação marginal nas análises clínica e microscópica. Conclusões: o sobrecontorno causado pelo laminado cerâmico não contribuiu para o acúmulo de placa na região cervical e para alterações no comportamento clínico periodontal, microbiológico e microscópico em relação aos mesmos dentes antes do tratamento restaurador(AU)

Purpose: quantify colonizing bacteria of the gingival sulcus, in parallel to the verify of the periodontal clinical behavior and the marginal adaptation of natural teeth restored with ceramic veneers without finish line. Methods: 73 ceramic veneers in monolithic lithium disilicate were cemented onto natural teeth without finish line and with the cervical margin positioned about 0.5mm inside the gingival sulcus. Gingival crevicular fluid (GCF) was collected for all teeth prior to cementation of the restoration (baseline) and 7, 180 and 365 days after cementation to quantify the bacteria S. mitis, P. intermedia and P. gingivalis through of real-time PCR. Clinically, visible plaque index (PIV), bleeding on probing (BOP), probing depth (PD), clinical attachment level (CAL), gingival recession (GR) and marginal adaptation were evaluated at baseline, 7, 15, 30, 60, 180 and 365 days. Replicas of the cervical region were obtained for analysis of marginal adaptation by scanning electron microscopy (SEM). Results: There was no statistically significant difference for PIV, PD, BOP in all periods analyzed (Anova, p>0.05). There were no records of CAL and RG. There was a statistically significant difference for S. mitis in the comparison between 180 and 365 days (Dunn, p=0.03). P. intermedia was not detected in any of the analyzed samples. There was no statistically significant difference for P. gingivalis at all study times (Friedman, p>0.05). All restorations had an alpha concept for marginal adaptation in clinical and SEM analysis. Conclusions: the overcontouring caused by ceramic veneers did not contribute to the accumulation of plaque in the cervical region and to changes in the clinical, microbiological and microscopic behavior in relation to the same teeth before the restorative treatment(AU)

Qualitative and quantitative molecular analysis of bacteria in root canals of primary teeth with pulp necrosis

Abstract Information about bacterial diversity, such as the number of each species in the root canals of primary teeth, contributes to improving our effective management of infections of endodontic origin in primary teeth. This study made a qualitative and quantitative assessment of the bacteria in the root canals of primary teeth with necrotic pulp, using the fluorescence in situ hybridization (FISH) technique. Thirty-one primary teeth with pulp necrosis from 31 children were evaluated using the FISH technique, to detect the presence and density of Aggregatibacter actinomycetemcomitans, Campylobacter rectus, Enterococcus faecalis, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Streptococcus, Streptococcus mutans, Streptococcus sobrinus, Tannerella forsythia and Treponema denticola. Descriptive measures explained the data related to density, and Student's t-test assessed the differences among the densities of each bacterium, according to signs and symptoms. The bacterial density was paired and correlated. All bacteria tested were detected and identified in all the samples. The average number of bacterial individuals from each species ranged from 1.9 x 108 cells/mL (S. mutans) to 3.1 x 108 cells/mL (F. nucleatum) (p > 0.05). The sum of the mean counts of each bacterium represented almost 80% of the entire microbial community. Patients with pain had significantly more T. denticola, and those with edema showed a greater density of Streptococcus and P. nigrescens (p < 0.05). This study revealed that all 12 bacteria evaluated were found in all primary teeth with pulp necrosis. There was no predominance among the species studied; all species had a similar number of individuals.

Research progress of correlation between periodontal pathogens and systemic diseases / 南方医科大学学报

Periodontal pathogens are the main pathogenic factor of periodontitis. Periodontal pathogens have a large variety of virulence factors such as lipopolysaccharide, fimbriae and proteases, which enables the pathogens to infect periodontal tissues and stimulate the secretion of inflammatory cytokines, causing chronic systemic inflammation. Periodontal pathogens may invade multiple systems such as the circulatory system, immune system, respiratory system and digestive system to cause systematic diseases. Recent studies have shown that periodontal pathogens may have close relations with systemic diseases such as cardiovascular disease, diabetes, rheumatoid arthritis, and cancer. Among the periodontal pathogens, can be found in atherosclerotic plaques to impairing the function of the vascular endothelium; may also increase the level of inflammatory factors such as TNF-α to promote insulin resistance and diabetes. Many of the periodontal pathogens such as , and can be detected in the synovial fluid of rheumatoid arthritis patients, suggesting their involvement in the pathogenesis of rheumatoid arthritis. may cause alterations in the intestinal microbiome in mice and promote the occurrence of intestinal tumors. Herein we review the recent progresses in the relationship between periodontal pathogens and systemic diseases.

Relationship of Oral Bacterial Load Over One Year of Smoking Cessation / 치위생과학회지

BACKGROUND: Smoking exerts an adverse effect on the periodontal tissue by reorganizing the ecosystem of oral microorganisms and is considered to be an important factor in the development of periodontal disease. Although cross-sectional studies on smokers and non-smokers have been attempted to investigate the microbial differences in periodontal oral cavity, only few studies have been conducted to investigate the changes in oral microorganisms during smoking cessation. The purpose of this study was to investigate the changes of bacteria in saliva and gingival crevicular fluid (GCF) over a period of one year among 11 smokers trying to quit smoking.METHODS: Eleven smokers trying to quit smoking visited the clinic at baseline, two weeks, two months, four months, six months, and 12 months to give saliva and GCF samples. The amounts of 16S rRNA, Porphyromonas gingivalis, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum subsp. nucleatum, Streptococcus mutans, and Streptococcus sobrinus in saliva and GCF were quantified using real-time polymerase chain reaction TaqMan probe assay. The results were analyzed by nonparametric statistical analysis using Friedman test and Spearman correlation coefficient.RESULTS: After cessation of smoking, the amounts of 16S rRNA corresponding to P. gingivalis, F. nucleatum, P. intermedia, and T. denticola in saliva decreased and then again increased significantly. The amount of F. nucleatum 16S rRNA in GCF decreased significantly after smoking cessation. Positive correlations were observed between 16S rRNA and F. nucleatum and between F. nucleatum and T. denticola in saliva and GCF.CONCLUSION: Even if the number of subjects in this study was small, we suggest that smoking cessation may reduce the total bacterial amount and F. nucleatum in GCF. However, the results regarding changes in the microbial ecosystem due to smoking or smoking cessation were inconsistent. Therefore, further in-depth studies need to be carried out.

Antibacterial Activity of Homeopathic Medications Lycopodium clavatum and Arsenicum album Against Periodontal Bacteria / Evaluación antibacteriana de los medicamentos homeopáticos sobre bacterias periodontales

Abstract There are several controversies regarding the efficacy of homeopathic substances; however, these remedies are used in many countries for the treatment of various pathological conditions. The purpose of this study was to evaluate the in vitro antibacterial activity of two homeopathic tinctures Arsenicum album (mineral extract) and Lycopodium clavatum (plant extract) on the periodontal bacteria Actinomyces israelii, Streptococcus sanguinis, Prevotella intermedia, Aggregatibacter actinomycetemcomitans and Phorphyromonas gingivalis (P. gingivalis). Materials and methods: Equal numbers of bacteria were seeded on agar plates containing enriched media with the homeopathic solutions at 1dH and 1cH dilutions. After 7 days of incubation under anaerobic conditions, colony forming units (CFUs) were counted. The antibacterial effect was calculated based on the total number of CFUs observed on non-tincture containing agar, and on the tincture containing plates. Results: No visible growth of any of the strains was observed on the plates containing Arsenicum album at any of the dilutions tested. In contrast, when Lycopodium clavatum at 1cH dilution was tested, only P. gingivalis was susceptible to this compound. Conclusions: The results suggest that the mineral extract tincture had a greater antibacterial activity than the plant extract tincture, also Lycopodium clavatum preparation could be an effective inhibitor of periodontal pathogens bacteria such as P. gingivalis.

Resumen Se necesita un mayor número de estudios in vitro e in vivo para validar estos resultados.

Comparison of full-mouth scaling and quadrant-wise scaling in the treatment of adult chronic periodontitis

Abstract In the search for the ideal treatment of periodontal disease various non-surgical techniques should be considered. The objective of this study was to evaluate the efficacy of full-mouth scaling (FMS) by clinical and microbiological parameters. 670 individuals were evaluated with 230 subjects meeting the selection criteria and were divided into two groups; 115 subjects treated with FMS and 115 treated with weekly sessions of scaling and root planning (SRP). The patient population had a mean age of 51.67 years, with moderate chronic periodontitis. Subjects were evaluated prior to treatment (T1) and 90 days after execution of therapy (T2), with regards to: probing depth (PD), clinical attachment level (CAL), plaque index (PI), gingival index (GI), and microbial detection for the presence of Porphyromonas gingivalis (P.g.) and Prevotella intermedia (P.i.) by culture method and confirmed by biochemical tests. Subjects treated in the FMS group also rinsed with 0.12% chlorhexidine mouthwash for seven days following treatment. The results were analyzed using statistical Student's t-test and chi-square test. No statistically significant differences were observed for PD and CAL between T1 and T2 in both groups. For GI and PI significant difference was observed between the groups. For the evaluated microbial parameters was observed reduction of P.g. and P.i., but only for P.g. with a significant reduction in both groups. The full mouth scaling technique with the methodology used in this study provided improved clinical conditions and reduction of P.g. in subjects with moderate periodontitis, optimizing the time spent in the therapeutic execution.

Resumo Na busca do tratamento ideal da doença periodontal varias são técnicas não-cirúrgicas que podem ser consideradas. O objetivo deste estudo foi avaliar a eficácia da técnica de desinfecção total de boca (FMD, na sigla em Inglês) por parâmetros clínicos e microbiológicos. Foram avaliados 670 indivíduos com 230 indivíduos atendendo aos critérios de seleção e divididos em dois grupos; 115 indivíduos tratados com FMD e 115 tratados com sessões semanais de raspagem e alisamento corono radicular (SRP, na sigla em Inglês). A população avaliada tinha idade média de 51,67 anos, com periodontite crônica moderada. Os sujeitos foram avaliados antes do tratamento (T1) e 90 dias após a execução da terapia (T2), quanto à profundidade de sondagem (PS), nível de inserção clínica (NIC), índice de placa (IP), índice gengival (IG) e detecção microbiana da presença de Porphyromonas gingivalis (P.g.) e Prevotella intermedia (P.i.) por método de cultura e confirmada por testes bioquímicos. Os indivíduos tratados no grupo FMD também realizaram bochechos com clorexidina 0,12% durante sete dias após o tratamento. Os resultados foram analisados ​​utilizando o teste estatístico t de Student e o teste de qui-quadrado. Não foram observadas diferenças estatisticamente significativas para PS e NIC entre T1 e T2 em ambos os grupos. Para IG e IP observou-se diferença significativa entre os grupos. Para os parâmetros microbianos avaliados foi observada redução de P.g. e P.i., mas apenas para P.g. com uma redução significativa em ambos os grupos. A técnica FMD com a metodologia utilizada neste estudo proporcionou condições clínicas melhoradas e redução da P.g. Em indivíduos com periodontite moderada, otimizando o tempo gasto na execução terapêutica.

Real-time PCR quantification of 9 periodontal pathogens in saliva samples from periodontally healthy Korean young adults

PURPOSE: Few studies have examined periodontal pathogens from saliva samples in periodontally healthy young adults. The purposes of this study were to determine the prevalence of periodontopathic bacteria and to quantify periodontal pathogens in saliva samples using real-time polymerase chain reaction (PCR) assays in periodontally healthy Korean young adults under 35 years of age. METHODS: Nine major periodontal pathogens were analyzed by real-time PCR in saliva from 94 periodontally healthy young adults. Quantification of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum, Campylobacter rectus, Peptostreptococcus anaerobius, and Eikenella corrodens was performed by DNA copy number measurement. RESULTS: F. nucleatum and E. corrodens were detected in all subjects; the numbers of positive samples were 87 (92.6%), 91 (96.8%), and 90 (95.7%) for P. gingivalis, P. anaerobius, and C. rectus, respectively. Other pathogens were also detected in periodontally healthy subjects. Analysis of DNA copy numbers revealed that the most abundant periodontal pathogen was F. nucleatum, which was significantly more prevalent than all other bacteria (P < 0.001), followed by P. anaerobius, P. gingivalis, E. corrodens, C. rectus, and T. denticola. There was no significant difference in the prevalence of each bacterium between men and women. The DNA copy number of total bacteria was significantly higher in men than in women. CONCLUSIONS: Major periodontal pathogens were prevalent in the saliva of periodontally healthy Korean young adults. Therefore, we suggest that the development of periodontal disease should not be overlooked in periodontally healthy young people, as it can arise due to periodontal pathogen imbalance and host susceptibility.

The effects of different restorative materials on periodontopathogens in combined restorative-periodontal treatment

Abstract Objective The aim of the study was to evaluate the association between subgingival restorations and the target periodontopathogenic bacteria (Pg, Td and Pi) in subgingival biofilm during one year after combined restorative-periodontal treatment. Material and Methods Seventeen systemically healthy subjects, who were positive for the presence of three cervical lesions associated with gingival recessions in three different adjacent teeth, were included in the study. A total of 51 combined defects were treated with connective tissue graft plus a nanofilled composite resin (NCR+CTG), a resin-modified glass ionemer cement (RMGI+CTG) and a fluoride-releasing resin material with pre-reacted glass (PRG), called giomer (Giomer+CTG). Periodontal clinical measurements and subgingival plaque samples were obtained from all combined defects at baseline and at 6 and 12 months after the surgery. The number of bacteria were evaluated by the real-time polymerase chain reaction (qPCR) method. Results No statistically significant difference in the amount of DNA copies of Pg, Td and Pi was observed in any of the groups at any time points (p>0.05). In addition, there was no statistically significant difference in the amount of DNA copies of the bacteria at baseline and at 6 and 12 months postoperatively, regardless of treatment group (p>0.05). Conclusion This study suggests that subgingivally placed NCR, RMGI and giomer restorations can show similar effects on periodontopathogenic bacteria in the treatment of gingival recessions that are associated with noncarious cervical lesions (NCCLs).

Genome sequence of Prevotella intermedia SUNY aB G8-9K-3, a biofilm forming strain with drug-resistance

Abstract Prevotella intermedia has long been known to be as the principal etiologic agent of periodontal diseases and associated with various systemic diseases. Previous studies showed that the intra-species difference exists in capacity of biofilm formation, antibiotic resistance, and serological reaction among P. intermedia strains. Here we report the genome sequence of P. intermedia SUNY aB G8-9K-3 (designated ATCC49046) that displays a relatively high antimicrobial resistant and biofilm-forming capacity. Genome sequencing information provides important clues in understanding the genetic bases of phenotypic differences among P. intermedia strains.

Red fluorescence of oral bacteria is affected by blood in the growth medium / 대한구강보건학회지

OBJECTIVES: Dental plaque emits red fluorescence under a visible blue light near the ultra-violet end of the light spectrum. The fluorescence characteristics of each microorganism have been reported in several studies. The aim of this study was to evaluate changes in red fluorescence of oral microorganisms that is affected by blood in the culture media. METHODS: The gram-positive Actinomyces naeslundii (AN, KCTC 5525) and Lactobacillus casei (LC, KCTC 3109) and gram negative Prevotella intermedia (PI, KCTC 3692) that are known to emit red fluorescence were used in this study. Each bacterium was activated in broth and cultivated in different agar media at 37℃ for 7 days. Tryptic soy agar with hemin and vitamin K3 (TSA), TSA with sheep blood (TSAB), basal medium mucin (BMM) medium, and BMM with sheep blood (BMMB) were used in this study. Fluorescence due to bacterial growth was observed under 405-nm wavelength blue light using the quantitative light-induced fluorescence-digital (QLF-D) device. The red, green, and blue fluorescence values of colonies were obtained using image-analysis software and the red to green ratio (R/G value) and red to total RGB ratio (R/RGB value) were calculated for quantitative comparison. RESULTS: The QLF-D images of the AN, LC, and PI colonies showed red fluorescence in all media, but the fluorescence of all bacteria was reduced in TSA and BMM media, compared with in TSAB and BMMB media. Both the R/G and the R/RGB values of all bacteria were significantly reduced in growth media without blood (P<0.001). CONCLUSIONS: Based on this in vitro study, it can be concluded that red fluorescence of oral bacteria can be affected by growth components, especially blood. Blood-containing medium could be a significant factor influencing red fluorescence of oral bacteria. It can be further hypothesized that bleeding in the oral cavity can increase the red fluorescence of dental plaque.

Eficacia inhibitoria de ciprofloxacina ótica y doxiciclina sobre cepas dePrevotella intermedia: Estudio in vitro / Eficácia inibitória de ciprofloxacina otica e doxiciclina em cepas de Prevotella intermedia: Estudo in vitro / Inhibitory efficacy of ciprofloxacin otic and doxycycline on strains of Prevotella intermedia: In vitro study

La ciprofloxacina ha sido propuesta como una alternativa para la aplicación local en terapia complementaria del tratamiento periodontal. Su forma tópica (ótica) se presenta como una alternativa de bajo costo y eficaz contra periodontopatógenos. Objetivo: Determinar el efecto inhibitorio de la Ciprofloxacina ótica (CIP) y Doxiciclina (DOXI) sobre cepas de Prevote-llaintermedia (Pi). Materiales y métodos: Estudio experimental, in vitro. La muestra estuvo constituida por cultivos de Pi ATCC®25611 en Agar Mueller Hinton para determinar mediante el test de susceptibilidad antibiótica el halo de inhibición. Se colocaron discos de CIP-5ug/10ul (G1 n=10); discos de DOXI-5ug (G2 n=10); y, discos de suero fisiológico-10ul (G3 n=10). Luego de 24 horas de incubación se realizó la medición de halos utilizando Antibiotic Zone Scale (mm) con los pa-rámetros del BD BBL Sensi-Disc Antimicrobial Suceptibibility Test Disc., con los siguientes parámetros: DOXI: resistente < 12 mm, intermedio 13-15 mm, sensible > 16 mm; CIPO: resistente < 15 mm, intermedio 16-20 mm, sensible > 21 mm. Se utilizó la prueba U de Mann Whitney con un nivel de significancia del 5%. Resultados: Todos los halos de inhibición de ambos grupos presentaron valores superiores a los parámetros de sensibilidad (efectividad 100%). La mediana para do-xiciclina fue de 38 mm y de ciprofloxacina fue de 36 mm. Existió diferencia estadísticamente significativa para los grupos G1 y G2 al ser comparados con los valores de referencia (p < 0.05). Conclusión: La ciprofloxacina local demostró la misma eficacia que la doxiciclina sobre cepas de Prevotella intermedia.

Ciprofloxacin has been proposed as an alternative for local application in complementary therapy of periodontal treatment. Its topical (otic) form presents as a low cost and effective alternative against periodontopathogens. Objective: To determine the inhibitory effect of Ciprofloxacin otica (CIP) and Doxycycline (DOXI) on strains of Prevotella intermedia (Pi). Materials and methods: Experimental study, in vitro. The sample was consisted of cultures of Pi ATCC®25611 in Agar Mueller Hinton to determine by the antibiotic susceptibility test the inhibition halo. CIP-5ug / 10ul discs (G1 n = 10) were placed; DOXI-5ug discs (G2 n = 10); and, physiological saline discs-10ul (G3 n = 10). After 24 hours of incubation, halos were measured using Antibiotic Zone Scale (mm) with the parameters of the BD BBL Sensi-Disc Antimicrobial Suceptibibility Test Disc., With the following parameters: DOXI: resistant <12 mm, intermediate 13-15 mm, sensitive> 16 mm; CIPO: resistant <15 mm, intermediate 16-20 mm, sensitive> 21 mm. The Mann Whitney U test was used with a level of significance of 5%. Results: All the inhibition zones of both groups presented values higher than the sensitivity parameters (100% effectiveness). Median for doxycycline was 38 mm and ciprofloxacin was 36 mm. There was a statistically significant difference for the G1 and G2 groups when compared with the reference values (p <0.05). Conclusion: Local ciprofloxacin demonstrated the same efficacy as doxycycline on strains of Prevotella intermedia.

A ciprofloxacina tem sido proposta como alternativa para aplicação local em terapia complementar do tratamento perio-dontal. Sua forma tópica (ótica) é apresentada como uma alternativa eficaz e de baixo custo contra os periodontopatôgenos. Objetivo: Determinar o efeito inibitório da Ciprofloxacina ótica (CIP) e Doxycycline (DOXI) em cepas de Prevotella intermedia (Pi). Materiais e métodos: Estudo experimental, in vitro. A amostra esteve constituída por culturas de Pi ATCC®25611 em Agar Mueller Hinton para determinar pelo teste de susceptibilidade aos antibióticos o halo de inibição. Foram colocados discos CIP-5ug / 10ul (G1 n = 10); Discos DOXI-5ug (G2 n = 10); e, discos solução salina - 10ul (G3 n = 10). Após 24 horas de incubação, os halos foram medidos usando a Antibiotic Zone Scale (mm) com os parâmetros do BD BBL Sensi-Disc Antimicrobial Suceptibility. Com os seguintes parâmetros: DOXI: resistente < 12 mm, intermediário 13-15 mm, sensível > 16 mm; CIPO: resistente < 15 mm, intermediário 16-20 mm, sensível > 21 mm. O teste U de Mann Whitney foi utilizado com um nível de significância de 5%. Resultados: Todos halos de inibição de ambos os grupos apre-sentaram valores superiores aos parâmetros de sensibilidade (100% de eficácia). A mediana da doxiciclina foi de 38 mm e a ciprofloxacina foi de 36 mm. Houve diferença estatisticamente significativa para os grupos G1 e G2 quando comparados com os valores de referência (p < 0,05). Conclusão: A ciprofloxacina local demonstrou a mesma eficácia que a doxiciclina em cepas de Prevotella intermedia.

Antimicrobial Effect of Coptidis rhizome Extract against Mutans Streptococci and Periodontopathogens

The purpose of the study was to investigate the antimicrobial activity of the methanol extract of Coptidis rhizome against the type strains of cariogenic bacteria, Streptococcus mutans and Streptococcus sobrinus, and the periodontopathogens, Porphyromonas gingivalis, Prevotella intermedia, Treponema denticola and Aggregatibacter actinomycetemcomitans. The antimicrobial activities of the crude extract and the methanol extract fractions of Coptidis rhizome separated by silica gel chromatography were evaluated by determining the minimal bactericidal concentration (MBC) values, using the microdilution method. The cell viability test of the extracts of Coptidis rhizome on the KB cells was also studied by methyl thiazolyl tetrazolium (MTT) assay. Our results showed that the 11th fraction (F11) of the methanol extract had the greatest antimicrobial activity against the tested bacteria, with no associated cytotoxicity on the KB cells, upto a concentration of 50 microg/ml. These results suggest that the silica gel chromatography fraction F11 of the methanol extract of Coptidis rhizome, could be useful in the development of oral hygiene products as an antimicrobial agent for the prevention of dental caries and periodontal diseases.

Development of Quantitative Real-Time PCR Primers for Detection of Prevotella intermedia

Prevotella intermedia-specific quantitative real-time PCR (qPCR) primers were previously designed based on the nucleotide sequences of RNA polymerase beta-subunit gene (rpoB). However, the several clinical strains isolated from Korean populations are not detectable by the qPCR primers. The purpose of this study was to develop new P. intermedia-specific qPCR primers based on the rpoB. The specificity of the primers was determined by conventional PCR with 12 strains of P. intermedia and 52 strains (52 species) of non-P. intermedia bacteria. The sensitivity of primers was determined by qPCR with serial dilutions of the purified genomic DNAs (40 ng to 4 fg) of P. intermedia ATCC 25611T. The data indicated that only P. intermedia strains were detected by the P intermedia-specific qPCR primers (RTPiF2/RTPiR2); in addition, as little as 40 fg of P. intermedia genomic DNA could be detected. These results suggest that these qPCR primers are useful in detecting P. intermedia from the bacterial infectious lesions including dental plaque and oral tissue lesions.

Osteoblast integration of dental implant materials after challenge by sub-gingival pathogens: a co-culture study in vitro / 国际口腔科学杂志·英文版

Sub-gingival anaerobic pathogens can colonize an implant surface to compromise osseointegration of dental implants once the soft tissue seal around the neck of an implant is broken. In vitro evaluations of implant materials are usually done in monoculture studies involving either tissue integration or bacterial colonization. Co-culture models, in which tissue cells and bacteria battle simultaneously for estate on an implant surface, have been demonstrated to provide a better in vitro mimic of the clinical situation. Here we aim to compare the surface coverage by U2OS osteoblasts cells prior to and after challenge by two anaerobic sub-gingival pathogens in a co-culture model on differently modified titanium (Ti), titanium-zirconium (TiZr) alloys and zirconia surfaces. Monoculture studies with either U2OS osteoblasts or bacteria were also carried out and indicated significant differences in biofilm formation between the implant materials, but interactions with U2OS osteoblasts were favourable on all materials. Adhering U2OS osteoblasts cells, however, were significantly more displaced from differently modified Ti surfaces by challenging sub-gingival pathogens than from TiZr alloys and zirconia variants. Combined with previous work employing a co-culture model consisting of human gingival fibroblasts and supra-gingival oral bacteria, results point to a different material selection to stimulate the formation of a soft tissue seal as compared to preservation of osseointegration under the unsterile conditions of the oral cavity.

Antibacterial effect of the carboxymethyl chitosan zinc peptide on several periodontal pathogens in vitro / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To determine the antimicrobial effects of carboxymethyl chitosan zinc (CMC-Zn(+)) and CMC-Zn(+)-peptide (CMC-Zn(+)-P) on four kinds of periodontal pathogens.</p><p><b>METHODS</b>Dilution method was used to determine the minimum inhibitory concentration (MIC) of CMC-Zn(+) for Porphyromonas gingivalis (Pg), Actinobacillusactinomycetemcomitans (Aa), Prevotella intermedia (Pi) and Actinomyces viscosus (Av). The antimicrobial characters of CMC-Zn(+) and CMC-Zn(+)-P on these four kinds of pathogens were evaluated by disk diffusion method.</p><p><b>RESULTS</b>The MIC of CMC-Zn(+) for Pg, Aa, Pi and Av was 0.312 5%, 0.156 25%, 0.156 25% and 0.078 125% respectively. Significant antimicrobial effects were improved along with the increased concentration of CMC-Zn(+) and CMC-Zn(+)-P (P < 0.01), while the best antimicrobial concentrations of both CMC-Zn(+) and CMC-Zn(+)-P were 5%. The antimicrobial effect of CMC-Zn(+)-P was better than that of CMC-Zn(+) on the same kind of periodontal pathogen (P < 0.01). CMC-Zn(+)-P showed different antimicrobial effects on the four periodontal pathogens (Av > Aa > Pi or Av > Aa > Pg, P < 0.01).</p><p><b>CONCLUSIONS</b>CMC-Zn(+) and CMC-Zn(+)-P have inhibition effect on Pg, Aa, Pi and Av, and the best antibacterial concentration was 5%. The CMC-Zn(+)-P has better antibacterial effect than CMC-Zn(+) on Pg, Aa, Pi and Av.</p>

Anti-inflammatory, Antioxidant and Antimicrobial Effects of Artemisinin Extracts from Artemisia annua L

The anti-inflammatory, antioxidant, and antimicrobial properties of artemisinin derived from water, methanol, ethanol, or acetone extracts of Artemisia annua L. were evaluated. All 4 artemisinin-containing extracts had anti-inflammatory effects. Of these, the acetone extract had the greatest inhibitory effect on lipopolysaccharide-induced nitric oxide (NO), prostaglandin E2 (PGE2), and proinflammatory cytokine (IL-1beta , IL-6, and IL-10) production. Antioxidant activity evaluations revealed that the ethanol extract had the highest free radical scavenging activity, (91.0+/-3.2%), similar to alpha-tocopherol (99.9%). The extracts had antimicrobial activity against the periodontopathic microorganisms Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum subsp. animalis, Fusobacterium nucleatum subsp. polymorphum, and Prevotella intermedia. This study shows that Artemisia annua L. extracts contain anti-inflammatory, antioxidant, and antimicrobial substances and should be considered for use in pharmaceutical products for the treatment of dental diseases.

Prevotella intermedia: uma breve revisão / Prevotella intermedia: A brief review

Prevotella intermedia é um microrganismo anaeróbio Gram negativo, considerado um dos agentes etiológicos de doenças periodontais. Tendo em vista que o biofilme subgengival é um dos seus principais habitats, qualquer quebra na homeostase do meio pode favorecer seu desenvolvimento e consequente dano aos tecidos atingidos. Diversas pesquisas vêm sendo realizadas a fim de identificar o papel deste microrganismo nas doenças periodontais. Assim, este trabalho teve por objetivo revisar a literatura para coletar informações sobre as características e efeitos de Prevotella intermedia no contexto saúde-doença periodontal. A busca foi realizada nas bases de dados PubMed e Lilacs, bem como através de uma busca aleatória. De um total de 47 artigos incluídos, alguns principais resultados puderam ser destacados: Prevotella intermedia está fortemente associada ao quadro de doenças periodontais e suas características favorecem o acometimento e intensificação de doenças diversas. Além disso, este microrganismo é capaz de produzir pigmentos e uma variedade de enzimas as quais favorecem danos aos tecidos periodontais. Existem várias características que aumentam a patogenicidade e virulência deste microrganismo, como por exemplos a sua cápsula polissacarídica, a presença de enzimas protetoras contra o oxigênio, a presença e produção de proteases, além da estimulação de fatores de crescimento no hospedeiro. Não há consenso na literatura acerca da relação de causa-efeito entre presença de Prevotella intermedia e diabetes. Contudo, este microrganismo parece estar envolvido no quadro de pacientes HIV positivo. Por fim, é importante salientar que Prevotella intermedia pode adquirir resistência a alguns tipos de antibióticos, e, por isso, o seu controle é um desafio à clínica atual...

Prevotella intermedia is a Gram negative anaerobic microorganism, considered one of the etiologic agents of periodontal diseases. Taking into account that the subgingival biofilm is its major habitat, any break of the environment homeostasis may lead to the development of the microorganism and consequent damage to the surrounding tissues. Several researches have been conducted in order to identify the role of this microorganism in the periodontal diseases. Thus, this study aimed to review the literature in an attempt to collect information about the characteristics and effects of Prevotella intermedia in the periodontal healthdisease context. The search was performed in the PubMed and Lilacs databases, as well as under a random search. From a total of 47 included articles, some major results could be drawn: Prevotella intermedia is strongly related to the occurrence of periodontal diseases, and its characteristics lead to the incidence of several diseases. Moreover, this microorganism is able to produce pigments and a variety of enzymes, which potentiate the damage of periodontal tissues. There are several characteristics that increase the pathogenicity and virulence of this microorganism, including the polyssacharidic capsule, the presence of protective enzymes against oxygen, the presence and production of proteases, as well as the stimulation of grow factors in the host. There is no consensus in the literature regarding the cause-effect relationship between the presence of Prevotella intermedia and diabetes. However, this microorganism seems to be involved in the occurrence of HIV-positive people. Lastly, it is worth to mention that Prevotella intermedia may acquire strength to some antibiotic types, and, therefore, its control is a challenge to the current clinic...