• Journal of Periodontal and Implant Science
• /
• v.29 no.3
• /
• pp.663-676
• /
• 1999

Porphyromonas gingivalis, a Gram negative, anaerobic, asaccharolytic rod, is one of the most frequently implicated pathogens in human periodontal disease and has a requirement for hemin for growth. A 30 kDa (heated 24 kDa) hemin-binding protein whose expression is both hemin and iron regulated has recently been purified and characterized in this oral pathogen. This study has identified a hemin-binding P. gingivalis protein by expression of a P. gingivalis genomic library in Escherichia coli, a bacterium which does not require or transport exogenous hemin. A library of genomic DNA fragments from P. gingivalis was constructed in plasmid pUC18, transformed into Escherichia coli strain $DH5{\alpha}$ , and screened for recombinant clones with hemin-binding activity by plating onto hemin-containing agar. Of approximately 10,000 recombinant E. coli colonies screened on LB-amp-hemin agar, 10 exhibited a clearly pigmented phenotype. Each clone contained various insert DNA. The Hind III fragment transferred to the T7 RNA polymerase/promoter expression vector system produced a sligltly smaller (21 kDa) protein, a precursor form, immunoreactive to the antibody against the 24 kDa protein, suggesting that the cloned DNA fragment probably carried an entire gene for the 24 kDa hemin-binding protein.

• International Journal of Oral Biology
• /
• v.46 no.3
• /
• pp.99-104
• /
• 2021

Associations between periodontal infection and cardiovascular disease have been documented. Porphyromonas gingivalis is a well-established periodontal pathogen, and tissue factor (TF) is a key initiator of the coagulation cascade. In this context, P. gingivalis has been reported to enhance TF expression in human endothelial cells. The present study investigated the underlying mechanisms of TF induction by P. gingivalis in human umbilical vein endothelial cells. P. gingivalis increased TF expression in a dose- and time-dependent manner. Not only live bacteria but also glutaraldehyde-fixed bacteria increased TF expression to the same extent. However, sonicates of P. gingivalis did not induce TF expression. Cytochalasin D and SMIFH2, which are inhibitors of actin polymerization and actin nucleation, respectively, inhibited the TF expression induced by P. gingivalis. Finally, TF production was decreased or increased in the presence of various signaling inhibitors, including mitogen-activated protein kinases. These results suggest that P. gingivalis induces endothelial TF expression by a bacterial internalization-dependent mechanism and through diverse signal transduction mechanisms.

• Journal of Periodontal and Implant Science
• /
• v.30 no.2
• /
• pp.419-429
• /
• 2000

Prevotella intermedia has been implicated as a potent pathogen in many kinds of periodontal, pulpal and periapical diseases. However, it has been isolated from periodontally healthy adults and from edentulous children as well. The intraspecies heterogeneity of Prevotella intermedia has been demonstrated in early studies and finally Shah & Gharbia confirmed the existence of 2 DNA homology groups and proposed dividing Prevotella intermedia into 2 species, Prevotella intermedia and Prevotella nigrescens. This study was designed to examine the frequency of Prevotella intermedia and Prevotella nigrescens in diseased periodontal pockets and healthy gingival sulcus of Korean people by PCR based on 16s ribosomal DNA sequence. One hundred adults who had adult periodontitis but not taken any periodontal treatment or antibiotics during previous 6 months and 50 adults who had healthy periodontal tissue were selected for this study. The sulcular fluid was collected into VMGA by sterilized paper point and diluted to 1,000 times in anaerobic chamber. $100{\mu}{\ell}$ of sample was cultured in $37^{\circ}C$ for 10 days. Among the bacterial colonies, BPB were selected and cultured in BHI broth and then Prevotella intermedia was identified through Gram staining and biochemical test. Identified Prevotella intermedia was cultured again and centrifuged. DNA was extracted from the pellet using several reagents. PCR was performed by previously designed primer. The results were followed. 1. BPB were isolated from 39 of 100 samples of diseased periodontal pockets(39%). 2. Prevotella intermedia was identified from 24 of 39 BPB samples. 3. Among 24 Prevotella intermedia, 21 were confirmed as Prevotella inter - media(87.5) and 2 were confirmed as Prevotella nigrescens(8.33%). 4. BPB were isolated from 9 of 50 samples of periodontally healthy patients. Among them only two were identified as Prevotella intermedia, that is, one was confirmed as Prevotella intermedia and the other was Prevotella nigrescens.

• Journal of Periodontal and Implant Science
• /
• v.33 no.3
• /
• pp.373-382
• /
• 2003

The successful use of osseointegrated implants to replace missing teeth has been demonstrated for both the completely and the partially edentulous patients. Many studies have confirmed an excellent long-term prognosis. The successful outcome of any implant procedure is surely dependent on the interrelationship of the various components that includes the following: biocompatibility of the implant material, macroscopic and microscopic nature of the implant surface, the status of the implant bed in both a health(noninfected) and a morphologic(bone quality) context, the surgical technique, the undisturbed healing phase, the subsequent prosthetic design, and long-term loading phase. Periodontally compromised patients have poor status of the implant bed and periodontal pathogen. No longitudinal data are available whether these factors affect the prognosis of implants. In this study, 102 machined $Br{{\aa}}nemark$ implants are inserted to analyze the success rate of 1-4 years and marginal bone loss in 49 chronic periodontitis patients. The following conclusions could be drawn from this study. 1. The cumulative success rate of implants at the 4-year of loading was 95.10%. 2. 5 failed implants have been removed. One implant have been removed due to infection, two implants were removed due to failure of osseointegration. and other two implants were removed due to mechanical failure caused by over-loading. 3. Mean marginal bone loss from the time of loading was 0.94mm at first year, 1.12mm at second year, 1.25mm at third year. These results suggest that implant therapy is good treatment modality in chronic periodontitis patients, and periodontal treatment including oral hygiene program is completed prior to insertion of implants.

• Journal of Korean society of Dental Hygiene
• /
• v.17 no.1
• /
• pp.13-25
• /
• 2017

Objectives: The purpose of this study was to compare colony forming unit (CFU) method and multiplex real-time polymerase chain reaction (MRT-PCR) method for accurate quantitative analysis of bacteria. Methods: We compared the CFU method and the MRT-PCR method, which are still used in Korea, for Prevotella intermedius (P. intermedius), a periodontal disease pathogen selected by MRT-PCR, and Streptococcus mutans (S. mutans), a dental caries causative organism. The subjects of this study were 30 patients who visited the C dental hospital. Results: Total microorganisms in MRT-PCR method were significantly higher in both types of bacteria (p<0.05), since DNA of dead bacteria was also analyzed. This was because the periodontal dise(-) anaerobes, and even dead bacteria contain large amounts of toxic substances called LPS in the extracellular membrane, and fimbriae and pili, which are motility structures, still remain as a strong toxic substance in periodontal tissue. Conclusions: Therefore, in terms of the total amount of bacteria found, the MRT-PCR method will be a useful technique for searching all the bacteria in the oral cavity including live bacteria, as well as sterilization.

• The Journal of the Korean dental association
• /
• v.52 no.12
• /
• pp.783-794
• /
• 2014

Since the introduction of non-thermal atmospheric pressure plasma in the field of the dentistry, numerous applications have been investigated. Especially with its advantages over existing vacuum plasma in terms of portability, low cost, and non-thermal damage, it can be directly applied in the oral cavity, giving number of potentials for dental application. First, possible application of non-thermal atmospheric pressure plasma in the field of dentistry is relation to dental caries and periodontal diseases. Teeth and alveolar bones are one of the strongest bony structures in our body, but it cannot be regenerated when they are damaged by dental caries or periodontal disease. Hence many studies to prevent such diseases have been carried out, though no perfect solution has been found yet. With recent studies of modifying surfaces through non-thermal atmospheric pressure application that can prevent attachment of bacteria, or studies on bactericidal effects of non-thermal atmospheric pressure plasma can be applied here to prevent oral pathogen and 'biofilm' attachment to the surface of teeth or directly eliminate the dental caries/periodontal disease causing germs. Secondly, non-thermal atmospheric pressure application will be useful on the surface of dental implant. It is well known that the success of dental implant surgery depends on the process known as 'osseointegration' that result from osteoblast attachment, proliferation and differentiation. As the application of non-thermal atmospheric pressure plasma on the surface of dental implant just before its introduction by the chair-side of dental surgery. Despite its long history, the generation of non-thermal atmospheric pressure plasma has been greatly increased with its application in dentistry.

• International Journal of Oral Biology
• /
• v.32 no.2
• /
• pp.75-78
• /
• 2007

The periodontal diseases and dental caries are major infectious diseases in oral cavity. Many of the preventive and therapeutic dental products contain the antimicrobial and anti-inflammatory components. But some of these antimicrobial have weak points such as coloration, burning sensation and insolubility in water. We have screened the therapeutic herbal extracts of the Plant Extract Bank for the antimicrobial activity on the major dental pathogens by growth inhibition assay. For the Porphyromonas gingivalis, 8 herbal extracts had an antimicrobial activity, 11 herbal extracts for the Prevotella intermedia, 43 herbal extracts for the Haemophilus actinomycetemcomitans and 61 herbal extracts for the Streptococcus mutans. Among these extracts, 6 herbal extracts had an antimicrobial activity for more than 3 species of dental pathogens. These extracts are Araliae Cordatae Radix, Crassirhizomae Rhizoma, Mori Radicis Cortex, Psoraleae Semen, Pini Ramulus and Sieges- beckiae Herba. All of effective extracts were CA group, ethanol extracts. Among these 6 herbal extracts, only Crassirhizomae is known to have a antibacterial effects. Therefore these herbal extracts have a possibility to be a candidate for a major antibacterial components in dental products.

• Journal of Periodontal and Implant Science
• /
• v.43 no.4
• /
• pp.147-152
• /
• 2013

Purpose: An animal periodontitis model is essential for research on the pathogenesis and treatment of periodontal disease. In this study, we have introduced a lipopolysaccharide (LPS) of a periodontal pathogen to the alveolar bone defect of experimental animals and investigated its suitability as a periodontitis model. Methods: Alveolar bone defects were made in both sides of the mandibular third premolar region of nine beagle dogs. Then, the animals were divided into the following groups: silk ligature tied on the cervical region of tooth group, Porphyromonas gingivalis LPS (P.g. LPS)-saturated collagen with silk ligature group, and no ligature or P.g. LPS application group as the control. The plaque index and gingival index were measured at 0 and 4 weeks postoperatively. The animals were then euthanized and prepared for histologic evaluation. Results: The silk ligature group and P.g. LPS with silk ligature group showed a significantly higher plaque index at 4 weeks compared to the control (P<0.05). No significant difference was found in the plaque index between the silk ligature group and P.g. LPS with silk ligature group. The P.g. LPS with silk ligature group showed a significantly higher gingival index compared to the silk ligature group or the control at 4 weeks (P<0.05). Histologic examination presented increased inflammatory cell infiltration in the gingival tissue and alveolar bone of the P.g. LPS with silk ligature group. Conclusions: An additional P.g. LPS-saturated collagen with silk ligature ensured periodontal inflammation at 4 weeks. Therefore, P.g. LPS with silk ligature application to surgically created alveolar bone defects may be a candidate model for experimental periodontitis.

• Journal of Periodontal and Implant Science
• /
• v.25 no.1
• /
• pp.89-98
• /
• 1995

P. intermedia are considered an important pathogen in adult periodontitis, rapidly progressing periodontitis, refractory periodontitis, pregnancy gingivitis, acute necrotizing ulcerative gingivitis, pubertal gingivitis. So far 2 DNA homology groups and 3 serotypes of P. intermedia have been reported but there is no data available as yet regarding genetic diversity for the species P. intermedia. The purpose of this study is to investigate, using bacterial DNA restriction endonuclease analysis, genetic diversity between individual strains of P. intermedia which are indistinguishable by serotyping and biotyping, occurrence of an intrafamilial transmission and genetic heterogeneity between P. intermedia strains isolated within a patient and within the same serotypes. The families who have had no systemic disease, no experience of periodontal treatment for the previous 1 year and no experience of antibiotics for the previous 6 months were selected and subgingival plaque was collected at 4 sites in each person and incubated in the anaerobic chamber. P. intermedia were identified by colony shape, gram stain, biochemical test, SK-I03(Sunstar Inc.) test and IIF using monoclonal antibody was perfomed for the determination of serotypes. P. intermedia strains were grown in BHI broth and whole genomic DNA was extracted and digested by restriction endonuclease. The resulting DNA fragments were separated by agarose gel electrophoresis, stained and photographed under UV. As the results of this study, intrafamilial vertial transmissions could be assessed in 2 families and horizintal transmissions in another 2 families. There were different DNA digest patterns within a patient, so P. intermedia showed that individuals could be colonized by multiple clonal types at anyone time. And different serotypes could be found within a patient and in the same serotype within a patient, obvius genetic heterogeneity could not be assessed. But in the same serotype in different famies, there were differences in the DNA digest patterns.

• Journal of Oral Medicine and Pain
• /
• v.24 no.4
• /
• pp.361-374
• /
• 1999

F. nucleatum is a gram-negative obligate anaerobe which is the principal and most frequent cause of gingival inflammation and is the predominant pathogen isolated in subsequent periodontal breakdown. It is also one of the most numerous bacteria found in subgingival plaque samples from healthy sites; its numbers are about 10-fold greater in plaque from periodontally diseased sites. The purpose of this study is to examine the effects of outer membrane(OM), outer membrane vesicle(OMV), and lipopolysaccharide(LPS) from F. nucleatum ATCC 25586 strain on the growth of human gingival fibroblasts and HOS 941 cells, and on the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression of mouse splenocytes. For the examination of cytotoxic effects, $TNF-{\alpha}$ production and $TNF-{\alpha}$ mRNA expression, the MTT assay, the ELISA and the RT-PCR were performed, respectively. All extracts of F. nucleatum tested were cytotoxic to both of human gingival fibroblasts and HOS 941 cells, and the significant difference of cytotoxic activity among the extracts was not observed. In the effects of these extracts on the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression of mouse splenocytes, all extracts of F. nucleatum tested also stimulated the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression, but the effects of the OM extracts on the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression were higher than those of the OMV and the LPS extracts. The pattern of the $TNF-{\alpha}$ mRNA expression was similar to that of the $TNF-{\alpha}$ production. These results indicate that F. nucleatum seems to contribute to the pathogenesis of periodontal diseases at least by its cytotoxicity, directly and its $TNF-{\alpha}$ production, indirectly.