Detection of oral spirochete (Treponema denticola) by modified Fontana staining technique from sub-gingival plaques of patients suffering from periodontal disease and to analyse the morphology of spirochetes with respect to the disease’s status

Background:A large proportion of fixed dose combinations (FDCs) are manufactured and used widely in Nepal. This study aimed to evaluate the FDCs and its utilization in medicine department of tertiary care hospital. Methods:A cross-sectional study was conducted for 50 days among admitted patients in the medicine department of tertiary care hospital, Nepal. A predesigned form was used to collect the data at the time of patient discharge. Only the oral FDCs were selected for study.Microsoft Excel 2007 was used for statistical analysis and data were presented as number and percentage in tabulated and figure forms.Results:Oral FDCs were used in 27.08% of admitted patients. A total of 295 FDCs were prescribed in 208 patients with 44 FDC items in 58 different brand names. Categorically, the most commonly used FDCs were of analgesics (34.24%) followed by antibiotics (25.76%) and vitamin supplements (22.71%). The 27.27% of FDCs prescribed contain more than two active pharmaceutical ingredients (APIs) up to nine and the highest number of APIs were found in vitamin supplements. All FDCs were prescribed in the brand names. The very few 2.27% and 4.55% of FDCs were prescribed from the essential medicine list of Nepal and world health organization, respectively.Conclusions:The use of FDCs listed in essential medicine list was very poor. Similarly, generic prescribing was also zero. The regulatory body must study the rationality of FDC before production, marketing, importing, and utilization in hospital.

jptsd123 / 口腔疾病防治

@#Esophageal cancer is one of the most lethal digestive system cancers, and its pathogenic factors have always been the focus of research. Recently, it has been found that microorganisms and their metabolites in the esophagus may also represent one of the pathogenic factors. Because of their continuity in anatomical structure, the oral cavity and esophagus have a certain correlation in terms of the composition of flora. In recent years, many scholars have studied the relationship between oral microorganisms and esophageal cancer to monitor changes in oral microorganisms as well as to diagnose and treat esophageal cancer more effectively. In this paper, the research status of oral microorganisms and esophageal cancer was reviewed. The Results of the literature review show that the diversity of bacteria in the esophagus is affected by oral flora in terms of the occurrence and development of esophageal cancer. Among these bacteria, the periodontal red complex, which includes Porphyromonas gingivalis, forsythia and Treponema dentata, as well as common oral microorganisms, such as Streptococcus viridis and Fusobacterium nucleatum, are all related to the occurrence and development of esophageal cancer to a certain extent. At present, there are few studies on the mechanism of microorganisms and esophageal cancer, but scholars have found that lipopolysaccharides and endotoxins, the products of Gram-negative bacteria in the esophagus, may participate in the innate immune response of the host, and the relevant mechanism of action needs further study in order to find new targets for monitoring and treatment.

Advances in Treponema denticola and oral diseases / 口腔疾病防治

@#Treponema denticola is an important pathogenic Treponema pathogen in the human oral cavity. Early studies have found that Treponema denticola is closely related to the occurrence and development of periodontal diseases. With the development of technical methods in recent years, many studies have shown that Treponema denticola not only can participate in periodontal diseases through a variety of mechanisms but also can play an important role in the development of various oral diseases. Treponema denticola is detected in high concentrations in peri-apical diseases and peri-implant diseases, and its surface protein is also prevalent in oral tumor samples. This paper reviews the research progress of Treponema denticola in periodontal diseases, pulp peri-apical diseases, peri-implant diseases and oral tumors, and summarizes the relevant mechanisms. For example, Treponema denticola can cause immune regulation disorder, destroy the epithelial barrier, induce bone absorption, promote the occurrence and development of inflammation through a variety of surface proteins, including chymotrypsin-like protease complex (CTLP), major outer sheath protein (Mosp), Td92, and LOS. It can also escape complement-mediated killing effects through surface FhbB lipoproteins and promote the occurrence and development of oral tumors by regulating the tumor microenvironment. These theories provide a theoretical basis for further understanding the development of oral diseases, controlling the infection of Treponema denticola, and exploring more effective treatment strategies.

Diagnóstico molecular de microorganismos periodontopatógenos en pacientes alcohó-licos - fumadores con periodontitis crónica de la ciudad de Loja, Ecuador / Diagnóstico molecular de microrganismos periodontopatogênicos em pacientes taba-gistas com periodontite crônica na cidade de Loja, Equador / Molecular diagnosis of microorganisms periodontopathogens in alcoholic - smoking patients with chronic periodontitis of the city of Loja, Ecuador

Los pacientes alcohólicos y fumadores presentan mayor predisposición de desarrollar enfermedad periodontal. Objetivo: Determinar la presencia de los microorganismos periodontopatógenos: Porphyromona gingivalis, Treponema denticola, Tannerella forsythia, Fusobacterium nucleatum y Aggregatibacter actinomycetemcomitans mediante la Reacción en Ca-dena de la Polimerasa (PCR), en pacientes alcohólicos fumadores con periodontitis crónica. Materiales y métodos: El universo fue de 31 pacientes de 18 y 62 años de edad de sexo masculino alcohólicos y/o fumadores en los cuales fueron excluidos aquellos que consumían algún tipo de droga, dando una muestra de 23 pacientes con bolsas periodontales ≥ 6mm del Centro de Rehabilitación de la ciudad de Loja, Ecuador. Se realizó un examen periodontal completo y la toma de muestras en dos de los sitios más profundos de cada paciente. Las variables analizadas fueron: presencia de biofilm dental, sangrado al sondaje, profundidad al sondaje y nivel de inserción clínica. Los datos fueron analizados con la prueba de Kruskal Wallis con un nivel de significancia del 5%. Resultados: La periodontitis crónica estuvo en el 52,1% de los pacientes de 18 a 30 años, siendo más susceptibles los alcohólicos de riesgo y fumadores leves. El 91,04 % de alcohóli-cos y fumadores se encuentran asociados con la presencia de biofilm dental (p = 0,028) y en el diagnóstico molecular el 41,18% de los pacientes presentan más de 3 microorganismos (p = 0,039). Conclusión: Se evidenció la presencia de los periodontopatógenos estudiados en pacientes alcohólicos fumadores con periodontitis crónica.

Alcoholic and smoking patients have a greater predisposition to develop periodontal disease. Objective: To determine the presence of microorganisms periodontopathogens: Porphyromona gingivalis, Treponema denticola, Tannerella forsythia, Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans by Polymerase Chain Reaction (PCR), in alcoho-lic-smoking patients with chronic periodontitis. Materials and methods: The group was composed of 31 patients between the ages of 18 to 62-year-old, male alcoholics and/or smokers in which were excluded those who take some kind of drug, giving a sample of 23 patients with periodontal pockets ≥ 6mm from the Center of Rehabilitation of the city of Loja, Ecuador. A full periodontal examination and the sample taking were in two of the most profound sites of each patient. The variables analyzed were: the presence of dental biofilm, bleeding on probing, probing depth and clinical attachment level. The data were analyzed with the Kruskal Wallis Test with a significance level of 5%. Results: The chronic periodontitis was in the 52.1% of the patients between the ages of 18 to 30 years old, is more susceptible in risky alcoholics-smokers and low smokers. The 91.04 % of alcoholics and smokers are associated with the presence of dental biofilm (p = 0.028) and in the molecular diagnosis the 41.18% of patients exhibit more than 3 microorganisms (p = 0.039). Conclusion: That observed presence of the periodontopathogens in alcoholic-smoking patients with chronic periodontitis.

Os pacientes alcoólicos e fumantes apresentam uma possibilidade maior de desenvolver doença peridental. Objetivo: Determinar a presença do periodontopatógenos de microorganismos Porphyromona gingivalis, Treponema denticola, Fusobacterium nucleatum e Aggregatibacter actinomycetemcomitans por meio da Reação em Cadeia de Polimerasa (PCR),em pacientes alcoólicos e fumantes com periodontite crônico. Materiais e métodos: O universo pertenceu a 31 pacientes de 18 e 62 anos de idade de sexo masculino alcoólicos e/ou fumantes nos quais foram excluídos aqueles que consumiram algum tipo de droga, tendo uma amostra de 23 pacientes com bolsas peridentais. 6mm do Centro de Reabilitação da cidade de Loja, Equador. Um exame periodontal completo e a amostragem foram realizados em dois locais profundamente localizados de cada paciente. As variáveis analisadas eram: presença de biofilm dental, sangrou ao sondaje, profundidade ao sondaje e nível de suplemento clínico. Os dados foram analisados com o teste de Kruskal Wallis com um nível de significancia de 5%. Resultados: O periodontite crônico estava em 52,1% dos pacientes de 18 para 30 anos, enquanto sendo mais suscetível os alcoólicos de risco e fumantes ligeiros. 91,04% de alcoólicos e fumantes são associados com a presença de biofilm dental (p = 0028) e na diagnose 41,18% molecular dos pacientes apresenta mais de 3 microorganismos (p = 0039). Conclusões: A presença do periodontopatógenos foi comprovada nos pacientes alcoólicos e fumantes com periodontite crônico.

Treponema denticola in microflora of bovine periodontitis / Treponema denticola na microbiota da periodontite bovina

Periodontitis in cattle is an infectious purulent progressive disease associated with strict anaerobic subgingival biofilm and is epidemiologically related to soil management at several locations of Brazil. This study aimed to detect Treponema species in periodontal pockets of cattle with lesions deeper than 5mm in the gingival sulcus of 6 to 24-month-old animals considered periodontally healthy. We used paper cones to collect the materials, after removal of supragingival plaques, and kept frozen (at -80°C) up to DNA extraction and polymerase chain reaction (PCR) using T. amylovorum, T. denticola, T. maltophilum, T. medium and T. vincentii primers. In periodontal pocket, it was possible to identify by PCR directly, the presence of Treponema amylovorum in 73% of animals (19/26), T. denticola in 42.3% (11/26) and T. maltophilum in 54% (14/26). Among the 25 healthy sites, it was possible to identify T. amylovorum in 18 (72%), T. denticola in two (8%) and T. maltophilum in eight (32%). Treponema medium and T. vincentii were not detected over all 51 evaluated samples. The presence of Treponema amylovorum, T. maltophilum and, in particular, the widely recognized T. denticola in subgingival microflora brings an original and potencially important contribution in studies of the bovine periodontitis.

A periodontite bovina é um processo infeccioso purulento e progressivo associado à presença de biofilme subgengival anaeróbio estrito e epidemiologicamente relacionada ao manejo do solo em amplas áreas geográficas do Brasil. O trabalho teve por objetivo detectar espécies de Treponema presentes na bolsa periodontal de bovinos com lesões de profundidade maior que 5mm e do sulco gengival de animais com idade de 6 a 24 meses e considerados periodontalmente sadios. Os materiais foram colhidos por meio de cones de papel, após a remoção do biofilme supragengival, e mantidos sob congelamento (-80°C) até a extração do DNA e realização da reação em cadeia da polimerase (PCR) com o emprego de iniciadores de T. amylovorum, T. denticola, T. maltophilum, T. medium e T. vincentii. Na bolsa periodontal de 73% (19/26) dos animais foi possível detectar diretamente, pela PCR, a presença de Treponema amylovorum, de 42,3% (11/26) T. denticola e de 54% (14/26) T. maltophilum. Dos 25 sítios sadios, em 18 (72%) foi possível identificar T. amylovorum, em dois (8%) T. denticola e em oito (32%) T. maltophilum. Treponema medium e T. vincentii não foram detectados nas 51 amostras avaliadas. A presença de Treponema amylovorum, T. maltophilum na microbiota subgengival, e em especial do amplamente reconhecido periodontopatógeno T. denticola, traz uma contribuição original de importância potencial nos estudos da periodontite bovina.

Coexistence of Porphyromonas gingivalis, Tannerella forsythia and Treponema denticola in the Red Bacterial Complex in Chronic Periodontitis Subjects / Coexistencia de Porphyromonas gingivalis, Tannerella forsythia y Treponema denticola en el Complejo Rojo Bacteriano en Sujetos con Periodontitis Crónica

Previous reports showed that periodontitis is associated with different microorganisms rather than individual periodontopathogens in the dental biofilm. The purpose of the current study was to evaluate the coexistence and relationship among Porphyromonas gingivalis, Tanerella forsythia, and Treponema denticola in the red complex, noting its association with the severity of periodontitis. In this cross sectional study, 96 subjects, aged 33 to 82 years (with 18 residual teeth) with chronic periodontitis who attended the dental clinics of the Universidad de Antioquia in Medellín, Colombia were invited to participate. The presence or absence of bleeding on probing and plaque were registered. Probing depth and clinical attachment level were measured at all approximal, buccal and lingual surfaces. Microbial sampling on periodontitis patients was performed on pockets >5 mm. The presence of P. gingivalis, T. forsythia, and T. denticola was detected by PCR using primers designed to target the respective 16S rRNA gene sequences. The coexistence of the three periodontopathogens was the most frequent (25 subjects). A statistically significant association between the three bacteria was observed (P. gingivalis and T. forsythia, P<0.0001; P. gingivalis and T. denticola, P=0.001; T. forsythia and T. denticola, P<0.0001). Similarly, the logistic regression analysis showed a significant association among periodontopathogens. The most relevant was observed between P. gingivalis and T. forsythia (OR=6.1). In conclusion, the present study found a significant association in the coexistence of P. gingivalis, T. forsythia and T. denticola, and they related strongly to clinical parameters of inflammation and periodontal destruction.

Reportes previos mostraron que la periodontitis se asocia con diferentes microorganismos en lugar de periodontopatógenos particulares en la biopelícula dental. El objetivo del presente estudio fue evaluar la coexistencia y relación entre Porphyromonas gingivalis, Tanerella forsythia y Treponema denticola en el complejo rojo, señalando su vinculación con la severidad de la periodontitis. En este estudio transversal, 96 sujetos de 33 a 82 años (con 18 dientes residuales) con periodontitis crónica que asistieron a las clínicas dentales de la Universidad de Antioquia en Medellín, Colombia fueron invitados a participar. Se registraron la presencia o ausencia de sangrado al sondaje y placa. La profundidad de sondaje y nivel de inserción clínica se midieron en todas las superficies proximales, bucal y lingual. El muestreo microbiano en pacientes con periodontitis se realizó en los bolsillos mayores a 5 mm. La presencia de P. gingivalis, T. forsythia, y T. denticola se detectó por PCR usando las bolsas periodontales diseñadas para dirigirse a las respectivas secuencias de genes 16S RNAr. La coexistencia de los tres periodontopatógenos fue la más frecuente (25 sujetos). Se observó una asociación estadísticamente significativa entre las tres bacterias (P. gingivalis y T. forsythia, P<0,0001; P. gingivalis y T. denticola, P=0,001; T. forsythia y T. denticola, P<0,0001). Del mismo modo, el análisis de regresión logística mostró una asociación significativa entre periodontopatógenos; la más relevantes se observó entre P. gingivalis y T. forsythia (OR=6,1). El presente estudio encontró una asociación significativa en la coexistencia de P. gingivalis, T. forsythia y T. denticola, y estuvieron fuertemente relacionadas a los parámetros clínicos de la inflamación y destrucción periodontal.

Periodontitis agresiva infantil en un niño de 4 años / Aggressive infantile periodontitis in a 4-year-old boy

Se define a la periodontitis agresiva como una forma agresiva y rápidamente destructiva de enfermedad, caracterizada por la pérdida severa del ligamento periodontal y hueso alveolar. Si bien la mayor prevalencia se ubica en la población adulta, se han registrado casos en niños y adolescentes. Se reporta el caso clínico de un niño de 4 años de edad que concurre a la consulta en el año 2010 por pérdida prematura de piezas dentarias. La evaluación de los exámenes de rutina y las radiografías con las que acudió permitieron descartar una periodontitis como manifestación de una enfermedad sistémica. Se confirma el diagnóstico de periodontitis infantil agresiva, por medio de examen microbiológico y ténica PCR. El tratamiento indicado fue terapia básica, antibioticoterapia y controles periódicos. Luego de tre años no hay signos clínicos de la enfermedad. El diagnóstico preciso, sumado a la motivación constante y los controles periódicos, hacen posible la detención del avance de la enfermedad...

Occurrence of periodontal pathogens among patients with chronic periodontitis

The aim of the present study was to evaluate the presence of the periodontal pathogens that form the red complex (Tannerella forsythia, Porphyromonas gingivalis and Treponema denticola) and Aggregatibacter actinomycetemcomitans in patients with chronic periodontitis. The sample consisted of 29 patients with a clinical and radiographic diagnosis of chronic periodontitis based on the criteria of the American Academy of Periodontology (3). Samples for microbiological analysis were collected from the four sites of greatest probing depth in each patient, totaling 116 samples. These samples were processed using conventional polymerase chain reaction, which achieved the following positive results: 46.6% for P. gingivalis, 41.4% for T. forsythia, 33.6% for T. denticola and 27.6% for A. actinomycetemcomitans. P. gingivalis and T. forsythia were more prevalent (p < 0.05) in periodontal pockets ≥ 8 mm. The combinations T. forsythia + P. gingivalis (23.2%) and T. forsythia + P. gingivalis + T. denticola (20.0%) were more frequent in sites with a probing depth ≥ 8 mm. Associations with the simultaneous presence of A. actinomycetemcomitans + P. gingivalis, A. actinomycetemcomitans + T. forsythia, P. gingivalis + T. forsythia and T. forsythia + T. denticola were statistically significant (p < 0.05). It was concluded that the red complex pathogens are related to chronic periodontitis, presenting a higher occurrence in deep periodontal pockets. Moreover, the simultaneous presence of these bacteria in deep sites suggests a symbiotic relationship between these virulent species, favoring, in this way, a further progression of periodontal disease.

Deteccão de diferentes microorganismos na periodontite crônica, da glicoproteína EMMPRIN (CD-147) e sua correlação com a produção de MMP-2 e MMP-9 / Detection of different microorganisms in chronic periodontitis, glycoprotein EMMPRIN (CD-147) and its correlation with production of MMP-2 and MMP-9

A periodontite é uma doença infecciosa caracterizada pela secreção de uma variedade de mediadores inflamatórios que levam a destruição dos tecidos de suporte dental, incluindo o osso alveolar e possível perda dos dentes, em associação com a infecção por múltiplas espécies de bactérias gram-negativas. Estima-se que mais de 400 espécies microbianas colonizam o biofilme dental e algumas das espécies bucais relacionadas à doença periodontal apresentem-se no biofilme subgengival incluindo Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola. Entretanto, outros microrganismos podem também estar relacionados a patologia desta doença, como Filifactor alocis, Prevotella tannerae e Candida albicans. Estes microrganismos juntamente com seus subprodutos como as endotoxinas liberadas no meio extracelular levam ao estímulo da glicoproteina Indutora de Metaloproteinases (EMMPRIN-CD147) a qual estimula a liberação de metaloproteinases da matriz (MMPs) pelas células do hospedeiro, por exemplo, fibroblastos e células endoteliais levando assim a destruição tecidual. Com o objetivo de detectar novos patógenos relacionados à doença periodontal como F. alocis, P. tannerae, C. albicans e T. denticola e também avaliar a glicoproteína EMMPRIN (CD- 147) e sua correlação com MMP-2 e MMP-9 em amostras de fluido subgengival de pacientes com periodontite crônica, foram coletadas amostras de fluido subgengival de sítios sadios e doentes de 20 indivíduos com periodontite crônica antes do tratamento periodontal básico, e após um período de 60 dias do tratamento foi realizada nova coleta de sítios sadios e doentes. Os respectivos DNAs foram extraídos e trechos do gene 16S foram amplificados e posteriormente realizados PCR convencional para a análise microbiológica dos microrganismos em estudo. Para a análise imunológica e quantificação de EMMPRIN-CD147, MMP-2 e MMP-9 foi utilizado a técnica de ELISA-Sanduíche. Os resultados demonstraram que o grupo doente mostrou valores significativamente elevados de T. denticola, F. alocis e P. tannerae quando comparado com sítios sadios. MMP-2 e MMP-9 foram detectados em concentrações elevadas, com redução estatisticamente significativa após o tratamento periodontal para MMP-2 e sem correlação com EMMPRIN

Periodontitis is an infectious disease characterized by the secretion of a variety of inflammatory mediators that lead to destruction of tooth supporting tissues, including the possible loss of alveolar bone and teeth, in association with infection with multiple species of gramnegative bacteria. It is estimated that more than 400 species of microorganisms colonize the biofilm and some oral species related to periodontal disease is present in the subgingival including Porphyromonas gingivalis, Tannerella forsythia and Treponema denticola. However, other organisms may also be related to pathology of this disease, as Filifactor allocis, Prevotella tannerae and Candida albicans. These microorganisms along with subproducts such as endotoxins released into the extracellular lead to the stimulation of metalloproteinase inducer glycoprotein (EMMPRIN, CD-147), which stimulates the release of MMPs by host cells, like fibroblasts and endothelial cells, thus leading to tissue destruction . The objective of this study was to detect new pathogens relationed with periodontal disease such as F. allocis, P. tannerae, C. albicans, T. denticola and also the glycoprotein EMMPRIN (CD-147) and its correlation with MMP-2 and MMP-9 in subgingival fluid samples of patients with chronic periodontitis. Fluid were collected from healthy and disease subgingival sites of 20 healthy individuals with chronic periodontitis before basic periodontal treatment, and after a period of 60 days of treatment was performed new collection of healthy and diseased sites. Their DNAs were extracted and portions of the 16S gene were amplified and subsequently performed conventional PCR for the microbiological analysis of the microorganisms this study. For immunological analysis and quantification of EMMPRIN (CD-147), MMP-2 and MMP-9 was used ELISA-Sandwich. Results demonstrated that the disease group showed significantly high amounts of T. denticola, F. alocis and P. tannerae when compared with health sites. MMP-2 and MMP-9 were detected in high concentrations with statistically significantly reduction after periodontal treatment to MMP-2, but without correlation with EMMPRIN

Identificación por reacción en cadena de la polimerasa (PCR) de microorganismos presentes en las infecciones orofaciales odontogénicas / Identification of bacteria associated with orofacial infections by polimerase chain reaction (PCR)

El objetivo del presente trabajo fue identificar la presencia de las bacterias, Porphyromonas gingivalis, Tannerella forsythensis y Treponema denticola, denominado complejo rojo; en grupo de tres, en parejas o sola una especie, presentes en las infecciones orofaciales odontogénicas (IOFO), mediante el método de la Reacción de la Polimerasa Reversa (PCR), en las muestras de abscesos de los pacientes que acudieron al Servicio de Odontoestomatologia del Hospital Nacional Cayetano Heredia y a la Clínica Estomatológica Central de la Universidad Peruana Cayetano Heredia. Se obtuvieron 35 muestras, en ninguna de ellas se identificó el complejo bacteriano integrado por P. gingivalis, T. forsythensis y T. denticola. La asociación entre P. gingivalis y el T. forsythensis fue la más frecuente con un 5,71%. Al menos uno de los integrantes del complejo bacteriano estuvo presente en el 48,57% (17 de 35) muestras. La P. gingivalis es la bacteria más prevalente del complejo bacteriano con el 40% (14 de 35) muestras, seguido de la T. forsythensisen un 11,43% (4 de 35) muestras. La P. gingivalis se identificó en un 42,86% (6 de 14) muestras del género femenino y 38,1% (8 de 21) muestras del género femenino. Este estudio ha demostrado por medio de la prueba de PCR, la prevalencia de tres especies bacterianas presentes en las infecciones orofaciales odontogénicas. Aunque la etiología de estos procesos es multifactorial, es muy importante considerar los resultados de este estudio para el manejo racional de las infecciones.

The objective of this investigation was to identify the prevalence of Porphyromonas gingivalis, Tannerella forsythensis and Treponema denticola forming the red complex, or as pairs or as single species in odontogenic infections, by mean of the Polymerase Chain Reaction (PCR). The samples were obtained from 35 oral and facial abscesses. In no case the red complex was found. The most frequent association found was P. gingivalis and T. forsythensis that represented 5.71%. At least, one specie was identified in 17 samples (48.57%). P. gingivalis was the most common bacteria representing 40% (14 out of 35 samples). T. forsythensis was identified in 11.43% (4 out of 35 samples). T. denticola was the less prevalent with only two cases (5.71%). P. gingivalis was demonstrated in 8 out of 21 males (38%) and in 6 out of 14 females (42.85%). This study has demonstrated by PCR the prevalence of three species of odontogenic bacteria as responsible of orofacial infections. Although the etiology of these proccesses is multifactorial, it is very important to consider the results of this study in the rational management of them. This investigation was carried out in the Stomatology Service of Cayetano Heredia National Hospital and in the Oral and Maxillofacial Surgery Service of the School of Dentistry of Cayetano Heredia Peruvian University.

Characterization of Binding of Treponema denticola to Immobilized Fibrinogen using the Fluorescent Fatty Acid Labeling Method

Treponema denticola is a gram-negative anaerobe that can cause periodontal disease. The adhesion of this bacterium to host tissues is considered to be the primary event in the colonization and infection of a host. Fibrinogen is generally found in damaged tissues resulting from periodontitis. The binding ability of T. denticola to fibrinogen may therefore be an important virulence factor in inducing periodontal diseases. It has been reported recently that oral spirochetes can be labeled with fluorescent fatty acids and we speculated that this labeling method could be used in an oral spirochete binding assay. The binding of several different strains of T. denticola to immobilized human fibrinogen was therefore tested using the fluorescent fatty acid labeling method. In the case of immobilized fibrinogen, the T. denticola ATCC 35405 strain showed saturable binding to immobilized fibrinogen. Indeed, all four different T. denticola strains tested in this experiment, T. denticola ATCC 35405, T. denticola ATCC 33520, T. denticola ATCC 35404 and T. denticola OTK showed binding to fibrinogen. The fluorescent fatty acid labeling method thus shows utility in binding assays for T. denticola, different strains of which can generally bind to immobilized fibrinogen.

Effect of sonicates of Treponema denticola on osteoblast differentiation / 대한치주과학회지

No abstract available.

The Effect of Treponema Denticola and Treponema Lecithinolyticum on Periodontal Ligament Cells / 대한치주과학회지

This study was investigated to observe the effect of Treponema denticola(TDC) and Treponema lecithinolyticum(TLC) on cultured human periodontal ligament cells. Several experiments were performed including MTT test for the inhibition effect of cell proliferation, LDH test for the cytotoxicity , gelatin zymography for the gelatinase activation and observation of cell morphology change using the phase-contrast microscopy. The results were as follows. 1. The effect of concentration on cell proliferation with time showed an inhibitory effect at high concentration(150microgram/well) for TLC and at low concentration( 9.4microgram/well ) for TDC. 2. The effect of time on cell proliferation with concentration showed an inhibitory effect at 150microgram/well on 2-day incubation for TLC and at 9.4microgram/well on 2-day incubation for TDC. 3. The effect of heat-treated TDC and TLC on the inhibition of cell proliferation showed the difference in the heat-treated group compared to the non-heat treated group for TDC, whereas no difference was found for TLC. 4. The morphological changes which were observed from the phase-contrast microscopy showed the difference in the test group compared to the control group. The loss of spindle-like appearance, cell-to-cell detachment and inhibition of cell proliferation were observed. 5. There was no difference of the cytotoxicity effect between the test group and the control group in the LDH test. 6. The active form of progelatinase A with molecular weight 72kDa was activated in both TDC and TLC on the gelatin zymography. Regarding to the above results, TDC and TLC have an effect on periodontal ligament cells by playing an inhibitory role in cell proliferation and appears to activate progelatinase A which degrades type IV collagen.

Effect of Sonicated Extract of Treponema Denticola on Osteoclast Differentiation / 대한치주과학회지

Alveolar bone destruction is a characteristic of periodontal disease. Treponema denticola are found in significantly increased numbers in the sites affected with periodontal disease. In order to clarify the role of T. denticola in destruction of alveolar bone in periodontal disease, this study was undertaken to determine the effect of sonicated extract of T. denticola on osteoclast differentiation in co-culture system of mouse bone marrow cells and calvaria cells. The ability of osteoclast formation was estimated by counting the number of tartrate resistant acid phosphatase(TRAP) positive cells. Sonicated extract of this bacteria stimulated osteoclast formation in a dose dependent manner(p<0.05). Indomathacin, an inhibitor of prostaglandin synthesis, decreased osteoclast formation induced by sonicated extract of this bacteria(p<0.05). Extract-induced osteoclast formation was decreased, when sonicated extract of bacteria was heated(p<0.05). These findings suggest that T. denticola induces osteoclast differentiation, and protein component of this bacteria and PGE2 may play an important role in this process.

The Effect of Sonicated Extracts of Treponema Denticola and Treponema Lecithinolyticum on the Cytokine Secretion and Matrix Metalloproteinase Activation of Gingival Fibroblast / 대한치주과학회지

This study was investigated to observe the effect of Treponema denticola cell sonicates(TDC) and Treponema lecithinolyticum cell sonicates(TLC) on cytokine secretion and matix metalloproteinase-2(MMP-2) activation of cultured human gingival fibroblast. Several experiments were performed including IL-1beta, IL-6 ELISA for the effect on the IL-1beta, IL-6 secretion of human gingival fibroblast. Also gelatinase zymography and gelatin dissolubility test for the activation of MMP-2 secreted by gingival fibroblast. The results were as follows. 1.The effect of TDC and TLC on IL-6 secretion of human gingival fibroblast showed statistically significant increase of IL-6 secretion in the TDC and TLC treated group compared to no treatment group(p<0.05) . 2.The amount of IL-1beta secretion was below the lower limit and there was no difference in the IL-1beta secretion of gingival fibroblast between TDC, TLC treated group and no treatment group. 3.The active form of pro MMP-2 with 72 kDa molecular weight was activated in both TDC and TLC treated group and clear band was appeared at 62kDa site on the zymography. 4.Gelatin dissolubility of MMP-2 secreted by gingival fibroblast was higher in TDC and TLC treated group compared to no treatment group(p<0.05). 5.In the TDC treated group, serine protease of T. denticola affect gelatin dissolubility. But in the TLC treated group gelatin was degraded by only MMP secreted by gingival fibroblast. Regarding to the above results, TDC and TLC have an effect on the IL-6 secretion increase of human gingival fibroblast and appears to activate pro MMP-2 which degrades collagen.