• 구강회복응용과학지
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• 제34권3호
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• pp.186-195
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• 2018

목적: 한국인에서 임플란트 주위 질환의 심도에 따른 미생물학적 차이를 알아보기 위해 real time Polymerase Chain Reaction(real-time PCR)법을 이용하여 5종의 치주세균의 정량적, 정성적 분석을 시행하였다. 연구 재료 및 방법: 임플란트가 식립된 총 60명의 환자를 치근단 방사선 사진 및 임상지수 검사를 통해 3군(건강군, 임플란트 주위 점막염군, 임플란트 주위염군)으로 나누었다. 멸균된 curette기구를 이용해 치은연하에서 미생물 샘플을 채취한 후 치주세균 5종에 관해 real time PCR을 시행하였고 comparative delta-CT method를 이용하여 분석한 후 미생물의 상대적 발현량을 비교하였다. 결과: Eikenella corrodens, Treponema denticola의 상대적 발현량은 임플란트 주위염 그룹에서 유의하게 높게 나타났다(P < 0.017). 반면 Fusobacterium nucleatum, Porphyromonas gingivalis의 상대적 발현량은 질환의 심도와는 관련 없이 건강한 임플란트 그룹에서 가장 높게 나타났다. Prevotella intermedia의 상대적 발현량은 건강한 임플란트 그룹에서 유의하게 낮게 나타났다(P < 0.017). 결론: 한국인의 임플란트 주위질환에서 대표적인 치주염 세균이 검출되었으나 치주염과 유사한 미생물학적 분포를 보이지는 않았다.

• Journal of Periodontal and Implant Science
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• 제45권5호
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• pp.178-183
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• 2015

Purpose: Elderly people are thought to be more susceptible to periodontal disease due to reduced immune function associated with aging. However, little information is available on the nature of immune responses against putative periodontal pathogens in geriatric patients. The purpose of this study was to evaluate the serum IgG antibody responses to six periodontal pathogens in geriatric subjects. Methods: The study population consisted of 85 geriatric patients and was divided into three groups: 29 mild (MCP), 27 moderate (MoCP), and 29 severe (SCP) chronic periodontitis patients. Serum levels of IgG antibody to Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, and Prevotella intermedia were measured by enzyme-linked immunosorbent assay (ELISA) and compared among the groups. Results: All three groups showed levels of serum IgG in response to P. gingivalis, A. actinomycetemcomitans, and P. intermedia that were three to four times higher than levels of IgG to T. forsythia, T. denticola, and F. nucleatum. There were no significant differences among all three groups in IgG response to P. gingivalis (P=0.065), T. forsythia (P=0.057), T. denticola (P=0.1), and P. intermedia (P=0.167), although the IgG levels tended to be higher in patients with SCP than in those with MCP or MoCP (with the exception of those for P. intermedia). In contrast, there were significant differences among the groups in IgG levels in response to F. nucleatum (P=0.001) and A. actinomycetemcomitans (P=0.003). IgG levels to A. actinomycetemcomitans were higher in patients with MCP than in those with MoCP or SCP. Conclusions: When IgG levels were compared among three periodontal disease groups, only IgG levels to F. nucleatum significantly increased with the severity of disease. On the contrary, IgG levels to A. actinomycetemcomitans decreased significantly in patients with SCP compared to those with MCP. There were no significant differences in the IgG levels for P. gingivalis, T. forsythia, T. denticola, and P. intermedia among geriatric patients with chronic periodontitis.

• 한국수정란이식학회지
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• 제21권3호
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• pp.241-245
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• 2006

본 연구는 젖소에 있어서 lipopolysaccharide의 처리가 번식 성적에 미치는 영향을 구명하기 위하여 2003년부터 2005년까지 3년간에 걸쳐 축산연구소 개방형 깔짚우사에서 사육중인 홀스타인 착유우 50두를 대상으로 처리구 및 대조구 각각 25두씩을 공시하였고 분만후 20일째에 1회에 한하여 Bacteroids helcogenes와 Fusobacterium varium으로부터 분리한 LPS $100{\mu}g$을 PBS 용액 35 ml에 희석하여 수정란 이식용 카테타로 자궁내에 주입한 후 발정 발현시 인공 수정을 실시하여 다음과 같은 결과를 얻었다. 1. LPS 처리후 첫 수정에 의한 수태율은 대조구 및 처리구가 각각 20.0% 및 56.0%로 처리구가 대조구에 비하여 높은 경향을 나타내었다. 2. LPS 처리후 2회 이상 수정에 의한 수태율은 대조구 및 처리구가 각각 40.0% 및 64.0%로 처리구가 대조구에 비하여 높은 경향을 나타내었다. 3. 수태된 개체들의 수태당 종부횟수는 대조구 및 처리구가 각각 $2.0{\pm}0.1$회 및 $1.2{\pm}0.4$회로 처리구가 대조구에 비하여 유의적으로 낮았다(p<0.05).

• Journal of Periodontal and Implant Science
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• 제44권6호
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• pp.266-273
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• 2014

Purpose: We previously reported that human serum significantly reduces the invasion of various oral bacterial species into gingival epithelial cells in vitro. The aims of the present study were to characterize the serum component(s) responsible for the inhibition of bacterial invasion of epithelial cells and to examine their effect on periodontitis induced in mice. Methods: Immortalized human gingival epithelial (HOK-16B) cells were infected with various 5- (and 6-) carboxy-fluorescein diacetate succinimidyl ester-labeled oral bacteria, including Fusobacterium nucleatum, Provetella intermedia, Porphyromonas gingivalis, and Treponiema denticola, in the absence or presence of three major serum components (human serum albumin [HSA], pooled human IgG [phIgG] and ${\alpha}1$-antitrypsin). Bacterial adhesion and invasion were determined by flow cytometry. The levels of intracellular reactive oxygen species (ROS) and activation of small GTPases were examined. Experimental periodontitis was induced by oral inoculation of P. gingivalis and T. denticola in Balb/c mice. Results: HSA and phIgG, but not ${\alpha}1$-antitrypsin, efficiently inhibited the invasion of various oral bacterial species into HOK-16B cells. HSA but not phIgG decreased the adhesion of F. nucleatum onto host cells and the levels of intracellular ROS in HOK-16B cells. N-acetyl-cysteine (NAC), a ROS scavenger, decreased both the levels of intracellular ROS and invasion of F. nucleatum into HOK-16B cells, confirming the role of ROS in bacterial invasion. Infection with F. nucleatum activated Rac1, a regulator of actin cytoskeleton dynamics. Not only HSA and NAC but also phIgG decreased the F. nucleatum-induced activation of Rac1. Furthermore, both HSA plus phIgG and NAC significantly reduced the alveolar bone loss in the experimental periodontitis induced by P. gingivalis and T. denticola in mice. Conclusions: NAC and the serum components HSA and phIgG, which inhibit bacterial invasion of oral epithelial cells in vitro, can successfully prevent experimental periodontitis.

• Journal of Periodontal and Implant Science
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• 제43권2호
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• pp.72-78
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• 2013

Purpose: The purpose of this study was to compare the phototoxic effects of blue light exposure on periodontal pathogens in both planktonic and biofilm cultures. Methods: Strains of Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, and Porphyromonas gingivalis, in planktonic or biofilm states, were exposed to visible light at wavelengths of 400.520 nm. A quartz-tungsten-halogen lamp at a power density of $500mW/cm^2$ was used for the light source. Each sample was exposed to 15, 30, 60, 90, or 120 seconds of each bacterial strain in the planktonic or biofilm state. Confocal scanning laser microscopy (CSLM) was used to observe the distribution of live/dead bacterial cells in biofilms. After light exposure, the bacterial killing rates were calculated from colony forming unit (CFU) counts. Results: CLSM images that were obtained from biofilms showed a mixture of dead and live bacterial cells extending to a depth of $30-45{\mu}m$. Obvious differences in the live-to-dead bacterial cell ratio were found in P. gingivalis biofilm according to light exposure time. In the planktonic state, almost all bacteria were killed with 60 seconds of light exposure to F. nucleatum (99.1%) and with 15 seconds to P. gingivalis (100%). In the biofilm state, however, only the CFU of P. gingivalis demonstrated a decreasing tendency with increasing light exposure time, and there was a lower efficacy of phototoxicity to P. gingivalis as biofilm than in the planktonic state. Conclusions: Blue light exposure using a dental halogen curing unit is effective in reducing periodontal pathogens in the planktonic state. It is recommended that an adjunctive exogenous photosensitizer be used and that pathogens be exposed to visible light for clinical antimicrobial periodontal therapy.

• Journal of Periodontal and Implant Science
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• 제50권3호
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• pp.171-182
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• 2020

Purpose: The aims of this study were to examine the salivary microbiota in conditions of periodontal health and disease and to explore microbial changes following nonsurgical periodontal treatment. Methods: Non-stimulated saliva samples were collected from 4 periodontally healthy participants at baseline and from 8 patients with chronic periodontitis at baseline and 3 months following nonsurgical periodontal therapy. The V3 and V4 regions of the 16S rRNA gene from the DNA of saliva samples were amplified and sequenced. The salivary microbial compositions of the healthy participants and patients with periodontitis prior to and following nonsurgical treatment of periodontitis were compared based on the relative abundance of various taxa. Results: On average, 299 operational taxonomic units were identified in each sample. The phylogenetic diversity in patients with periodontitis was higher than that in healthy participants and decreased following treatment. The abundance of the phylum Spirochaetes and the genus Treponema in patients with periodontitis was 143- and 134-fold higher than in the healthy control group, respectively, but decreased significantly following treatment. The species that were overabundant in the saliva of patients with periodontitis included the Peptostreptococcus stomatis group, Porphyromonas gingivalis, the Fusobacterium nucleatum group, Parvimonas micra, Porphyromonas endodontalis, Filifactor alocis, and Tannerella forsythia. The phylum Actinobacteria, the genus Streptococcaceae_uc, and the species Streptococcus salivarius group were more abundant in healthy participants than in those with periodontitis. There was a trend toward a decrease in disease-associated taxa and an increase in health-associated taxa following treatment. Conclusions: Our results revealed differences in the taxa of salivary microbiota between conditions of periodontal health and disease. The taxa found to be associated with health or disease have potential for use as salivary biomarkers for periodontal health or disease.

• International Journal of Oral Biology
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• 제36권3호
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• pp.109-116
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• 2011

Periodontitis results from the activation of host immune and inflammatory defense responses to subgingival plaque bacteria, most of which are gram-negative rods with lipopoly-saccharides (LPSs) in their cell walls. LPSs have been known to induce proinflammatory responses and recently it was reported also that they induce the expression of microRNAs (miRNAs) in host cells. In our current study therefore, we aimed to examine and compare the miRNA expression patterns induced by the LPSs of major periodontopathogens in the human gingival epithelial cell line, Ca9-22. The cells were treated with 1 ${\mu}g$/ml of E. coli (Ec) LPS or 5 ${\mu}g$/ml of an LPS preparations from four periodontopathogens Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Aggregatibacter actinomycetemcomitans (Aa), and Fusobacterium nucleatum (Fn) for 24 h. After small RNA extraction from the treated cells, miRNA microarray analysis was carried out and characteristic expression profiles were observed. Fn LPS most actively induced miRNAs related to inflammation, followed by Aa LPS, Pi LPS, and Ec LPS. In contrast, Pg LPS only weakly activated miRNAs related to inflammation. Among the miRNAs induced by each LPS, miR-875-3p, miR-449b, and miR-520d-3p were found to be commonly up-regulated by all five LPS preparations, although at different levels. When we further compared the miRNA expression patterns induced by each LPS, Ec LPS and Pi LPS were the most similar although Fn LPS and Aa LPS also induced a similar miRNA expression pattern. In contrast, the miRNA profile induced by Pg LPS was quite distinctive compared with the other bacteria. In conclusion, miR-875-3p, miR-449b, and miR-520d-3p miRNAs are potential targets for the diagnosis and treatment of periodontal inflammation induced by subgingival plaque biofilms. Furthermore, the observations in our current study provide new insights into the inflammatory miRNA response to periodontitis.

• Journal of Periodontal and Implant Science
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• 제43권4호
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• pp.183-190
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• 2013

Purpose: At present, information regarding periodontal disease in geriatric patients is scarce. The purpose of this study was to quantify the periodontal pathogens present in the saliva of Korean geriatric patients and assess the relationship between the bacterial levels and the periodontal condition. Methods: Six putative periodontal pathogens were quantified by using a real-time polymerase chain reaction assay in geriatric patient groups (>60 years) with mild chronic periodontitis (MCP), moderate chronic periodontitis (MoCP), and severe chronic periodontitis (SCP). The copy numbers of Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, and Prevotella intermedia were measured. Results: It was found that the bacterial copy numbers increased as the severity of the disease increased from MCP to SCP, except for P. intermedia. For P. intermedia, it was found that samples in the MCP group yielded the largest amount. It was also found that the quantities of P. gingivalis, T. forsythia, and T. denticola, the so-called "red complex" bacteria, were lower than those of F. nucleatum, A. actinomycetemcomitans, and P. intermedia in all of the samples. Conclusions: Collectively, the results of this study suggest that the levels of P. gingivalis, T. forsythia, F. nucleatum, and T. denticola present in saliva are associated with the severity of periodontal disease in geriatric patients.

• Journal of Periodontal and Implant Science
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• 제38권2호
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• pp.143-152
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• 2008

Purpose: Specific bacteria are believed to play an important role in chronic periodontitis. Although extensive microbial analyses have been performed from subgingival plaque samples of periodontitis patients, systemic analysis of subingival microbiota has not been carried out in a Korean population so far. The purpose of this study was to investigate the prevalence of 29 putative periodontal pathogens in Korean chronic periodontitis patients and evaluate which pathogens are more associated with Korean chronic periodontitis. Material and Methods: A total of 86 subgingival plaque samples were taken from 15 chronic periodontits(CP) patients and 13 periodontally healthy subjects in Korea. CP samples were obtained from the deepest periodontal pocket (>3 mm probing depth[PD]) and the most shallow periodontal probing site ($\leq$3 mm PD) in anterior tooth and posterior tooth, respectively, of each patient. Samples in healthy subjects were obtained from 1 anterior tooth and 1 posterior tooth. Polymerase chain reaction (PCR) of 16S ribosomal DNA (rDNA) of subgingival plaque bacteria was performed. Detection frequencies(% prevalence) of 29 putative periodontal pathogens were investigated as bacterium-positive sites/total sites. Results: With the exception of Olsenella profuse and Prevotella nigrescens, the sites of diseased patients generally showed higher prevalence than the healthy sites of healthy subjects for all bacteria analyzed. Tanerella forsythensis (B.forsythus), Campylobacter rectus, Filifactor alocis, Fusobacterium nucleatum, Porphyromonas endodontalis and Porphyromonas gingivalis were detected in more than 80% of sites with deep probing depths in CP patients. In comparison between the sites (deep or shallow PD) of CP patients and the healthy sites of healthy subjects, there was statistically significant difference(P<0.05) of prevalence in T.forsythensis (B.forsythus), C.rectus, Dialister invisus, F.alocis, P.gingivalis and Treponema denticola. Conclusion: Our results demonstrate that the four putative periodontal pathogens, T.forsythensis (B.forsythus), C.rectus, P.gingivalis and F.alocis are closely related with CP patients in the Korean population.

• Reproductive and Developmental Biology
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• 제30권3호
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• pp.195-199
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• 2006

본 연구는 후산정체 젖소에 있어서 Lipopolysaccharide(LPS)의 처리가 번식 성적에 미치는 영향을 구명하기 위하여 분만 후 12시간이 경과하여도 후산이 배출되지 않은 홀스타인 개체 33두 및 대조구 12두에 대하여 분만후 20일째에 LPS 100 ${\mu}g$을 자궁 내에 주입한 다음 7일후에 발정이 발현되었을 때 인공수정을 실시하여 다음과 같은 결과를 얻었다. 후산정체 젖소에 있어서 첫 수정에 의한 수태율은 대조구의 경우 공시한 12두가 모두 첫 수정에 의해 수태가 되지 않았고 처리구는 33두중 11두가 수태되어 33.3%의 수태율을 나타내었다. 후산정체 젖소에 있어서 대조구 및 처리구의 분만후 수태까지의 일수는 각각 $149.6{\pm}34.3$일 및 $53.0{\pm}12.5$일로 처리구가 대조구에 비하여 유의적(p<0.01)으로 단축되는 경향을 나타내었다. 후산정체 젖소에 있어서 대조구 및 처리구의 수태당 종부 횟수는 각각 $3.6{\pm}0.8$회 및 $2.1{\pm}0.3$회로 처리구가 대조구에 비하여 유의적(p<0.05)으로 낮은 경향을 나타내었다.