• Journal of Veterinary Science
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• v.25 no.4
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• pp.50.1-50.6
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• 2024

Importance: Canine extraction of large carnivores can pose significant risk due to extensive tissue damage during aggressive bone reduction. This report highlights a rare instance in which the use of a piezoelectric surgical unit (PSU) for maxillary canine extraction in a large carnivore resulted in successful outcomes with minimal tissue damage. Case presentation: A 10-year-old male African lion presented with decreased appetite because of bilateral maxillary canine fractures. Intraoral radiographs revealed enlarged root canals and periapical radiolucency of the fractured canines, leading to a diagnosis of periapical periodontitis and pulpitis. To extract the right maxillary canine, conventional method using hand instrument failed to achieve adequate luxation, necessitating the use of the flat blade of the PSU to sever the periodontal ligament. The left maxillary canine was extracted using PSU from the beginning, and the extraction time was markedly shortened by using PSU without additional alveolar bone damage or bleeding. Conclusion and Relevance: This case demonstrated that utilizing PSU for canine extraction in a lion resulted in periodontal ligament separation, reducing damage to the alveolar bone and shortening surgical time. It suggests the promising application of PSU in tooth extraction for large wild animals, indicating its potential significance in veterinary dentistry.

• 대한치주과학회:학술대회논문집
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• 2003.11a
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• pp.106-107
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• 2003

• International Journal of Oral Biology
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• v.37 no.3
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• pp.137-145
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• 2012

Aggregatibacter actinomycetemcomitans is the most important etiologic agent of aggressive periodontitis and can interact with endothelial cells. Monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) are chemokines, playing important roles in periodontal pathogenesis. In our current study, the effects of A. actinomycetemcomitans on the production of MCP-1 and IL-8 by human umbilical vein endothelial cells (HUVEC) were investigated. A. actinomycetemcomitans strongly induced the gene expression and protein release of both MCP-1 and IL-8 in a dose- and time-dependent manner. Dead A. actinomycetemcomitans cells were as effective as live bacteria in this induction. Treatment of HUVEC with cytochalasin D, an inhibitor of endocytosis, did not affect the mRNA up-regulation of MCP-1 and IL-8 by A. actinomycetemcomitans. However, genistein, an inhibitor of protein tyrosine kinases, substantially inhibited the MCP-1 and IL-8 production by A. actinomycetemcomitans, whereas pharmacological inhibition of each of three members of mitogen-activated protein (MAP) kinase family had little effect. Furthermore, gel shift assays showed that A. actinomycetemcomitans induces a biphasic activation (early at 1-2 h and late at 8-16 h) of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) and an early brief activation (0.5-2 h) of activator protein-1 (AP-1). Activation of canonical NF-${\kappa}B$ pathway ($I{\kappa}B$ kinase activation and $I{\kappa}B-{\alpha}$ degradation) was also demonstrated in these experiments. Although lipopolysaccharide from A. actinomycetemcomitans also induced NF-${\kappa}B$ activation, this activation profile over time differed from that of live A. actinomycetemcomitans. These results suggest that the expression of MCP-1 and IL-8 is potently increased by A. actinomycetemcomitans in endothelial cells, and that the viability of A. actinomycetemcomitans and bacterial internalization are not required for this effect, whereas the activation of protein tyrosine kinase(s), NF-${\kappa}B$, and AP-1 appears to play important roles. The secretion of high levels of MCP-1 and IL-8 resulting from interactions of A. actinomycetemcomitans with endothelial cells may thus contribute to the pathogenesis of aggressive periodontitis.

• Journal of Periodontal and Implant Science
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• v.38 no.sup2
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• pp.317-324
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• 2008

Purpose: Aggregatibacter actinomycetemcomitans was associated with localized aggressive periodontitis, endocarditis, meningitis, and osteomyelitis. The cytolethal distending toxin (CDT) of A. actinomycetemcomitans was considered as a key factor of these diseases is composed of five open reading frames (ORFs). Among of them, An enzymatic subunit of the CDT, CdtB has been known to be internalized into the host cell in order to induce its genotoxic effect. However, CdtB can not be localized in host cytoplasm without the help of a heterodimeric complex consisting of CdtA and CdtC. So, some studies suggested that CdtC functions as a ligand to interact with GM3 ganglioside of host cell surface. The precise role of the CdtC protein in the mechanism of action of the holotoxin is unknown at the present time. The aim of this study was to generate recombinant CdtC proteins expression from A. actinomycetemcomitans, through gene cloning and protein used to investigate the function of Cdt C protein in the bacterial pathogenesis. Materials and Methods: The genomic DNA of A. actinomycetemcomitans Y4 (ATCC29522) was isolated using the genomic DNA extraction kit and used as template to yield cdtC genes by PCR. The amplifed cdtC genes were cloned into T-vector and cloned cdt C gene was then subcloned to pET28a expression vector. The pET28a-cdtC plasmid expressed in BL21 (DE3) Escherichia coli system. Diverse conditons were tested to opitimize the expression and purification of functional CdtC protein in E. coli. Results: In this study we reconstructed CdtC subunit of A. actinomycetemcomitans Y4 and comfirmed the recombinant CdtC expression by SDS-PAGE and Western Blotting. The expression level of the recombinant CdtC was about 2% of total bacterial proteins. Conclusion: The lab condition of procedure for the purification of functionally active recombinant CdtC protein is established. The active recombinant CdtC protein will serve to examine the role of CdtC proteins in the host recognition and enzyme activity of CDT and investigate the pathological process of A. actinomycetemcomitans in periodontal disease.

• Journal of Periodontal and Implant Science
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• v.36 no.1
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• pp.113-123
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• 2006

연구배경 IgG에 대한 $Fc{\gamma}$ 수용기는 치주병인균에 대한 숙주 반응에 있어서 중요한 역할을 하는데, 이 중 $Fc{\gamma}RIIIa$는 NK 세포, 대식세포, 단핵구, ${\gamma}{\delta}T$세포에서 발현되며, EC2 도메인에서 158 아미노산 부위의 valine (V)-phenylalanine (F)의 유전자다형성을 보인다. $Fc{\gamma}RIIIb$는 특이적으로 중성구에 발현되는데, extracellular (ECl) Ig-like 도메인 내 4개의 아미노산 치환(substitutions)에 의한 NA1-NA2 유전자다형성을 보인다. 이 연구의 목적은 한국인에서 급진성 치주염 환자와 $Fc{\gamma}III$ 수용기의 유전자다형성과의 관련성을 알아보는 것이다. 연구방법 및 재료 치주적으로 건강한 90명 (대조군, 남자 64명, 여자 26명)과 서울대학교 치과병원 지주과에 내원하여 급진성 치주염으로 진단된 환자 43명 (aggressive periodontitis patients: AgP, 남자 30명, 여자 13명)을 대상으로 하였다. 모든 실험 대상자는 임상 실험에 대해 동의 하였고, 초진 시 전자 탐침(Florida Probe(R) Co. Gainesville, FL)을 이용하여 탐침 시 치주낭 깊이 (PPD), 임상부착수준 (CAL), 치태지수(PI), 탐침 후 출혈지수 (BOP)를 측정하였다. 또한 이들의 정맥혈에서 추출한 DNA를 PCR법, 전기영동법 등을 이용하여 $Fc{\gamma}RIIIa$, $Fc{\gamma}RIIIb$의 대립 유전자의 존재여부를 확인하였다. 이를 바탕으로 $Fc{\gamma}RIII$ 복합 유전형을 확인하여 각 군 간을 비교하였다. 연구 결과 1. $Fc{\gamma}RIIIa$에 대한 유전자다형성 연구 결과 대조군과 급진성 치주염 환자 군(AgP)사이에서는 대립 유전자 분포가 서로 유의성 있는 차이를 나타내었고 (P<0.05), $Fc{\gamma}RIIIb$에서는 유의성 있는 차이를 발견할 수 없었다. (P>0.05) 2. $Fc{\gamma}RIIIa$ 158F 대립형질이 급진성 치주염 환자에서 유의성 있게 많이 발견되어졌다. (P<0.05) 결론 이 연구를 통하여 $Fc{\gamma}RIIIa$ 유전자의 분석이 한국인의 급진성 치주염에 대한 감수성의 위험요소의 표지자로 활용할 수 있을 것으로 사료되며, 향 후 더 많은 환자를 대상으로 히는 추가 연구가 필요하다고 생각된다.

• Journal of Periodontal and Implant Science
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• v.38 no.sup2
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• pp.343-354
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• 2008

Purpose: Aggregatibacter actinomycetemcomitans is associated with localized aggressive periodontitis. It produces cytolethal distending toxin (CDT), which induces cell cycle arrest in the G2/M phase. The CDT holotoxin is composed of CdtA, CdtB, and CdtC. CdtB has structural homology to human DNase I and is an active component of the CDT complex acting as a DNase. In particular, the pattern homology seen in the CdtB subunit has been associated with specific DNase I residues involved in enzyme catalysis, DNA binding, and metal ion binding. So, to study the functions and regulation of recombinant CdtB, we made up a quantity of functional recombinant CdtB and tested it in relation to the metal ion effect. Materials and Methods: We constructed the pET28a-cdtB plasmid from A. actinomycetemcomitans Y4 by genomic DNA PCR and expressed it in the BL21 (DE3) Escherichia coli system. We obtained the functional recombinant CdtB by the refolding system using the dialysis method and then analyzed the DNase activity and investigated the metal ion effect from plasmid digestion. Results: The recombinant CdtB subunit was expressed as the inclusion bodies. We were able to obtain functional recombinant CdtB subunit using refolding system. We confirmed that our refolded recombinant CdtB had DNase activity and was influenced by the metal ions $Mg^{2+}$ and $Ca^{2+}$. Conclusion: We suggest that the factors influencing recombinant CdtB may contribute to CDT associated diseases, such as periodontitis, endocarditic, meningitis, and osteomyelitis.

• Journal of Periodontal and Implant Science
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• v.40 no.2
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• pp.49-55
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• 2010

Purpose: The purpose of this study was to preliminarily evaluate the influence of diabetes mellitus (DM) on periodontal tissue without establishment of periodontitis. Methods: Seven-week-old db/db mice were used for the diabetic experimental group and systematically healthy mice of the same age were used as controls. After 1 week of acclimatization, the animals were sacrificed for hard and soft tissue evaluation. The pattern of bone destruction was evaluated by stereomicroscope evaluation with alizarin red staining and radiographic evaluation by microscopic computerized tomography images. Histological evaluation was performed with hematoxylin and eosin stain for evaluation of soft tissue changes. Results: In both stereomicroscope evaluation and radiograph image analysis, aggressive form of bone destruction was observed in diabetic animals when compared to the systematically healthy controls. In histological evaluation, apical migration of junctional epithelium with slight inflammatory cell infiltration was observed with disarrangement of connective tissue fibers. Conclusions: Within the limits of this study, diabetic animals presented distortion in periodontal attachment and an aggressive bone loss pattern when compared to the healthy controls, suggesting that DM has an independent effect on periodontal tissue destruction irrespective of the presence or absence of periodontal disease.

• Journal of Periodontal and Implant Science
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• v.27 no.3
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• pp.533-547
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• 1997

In periodontics, much progress was made in the understanding of periodontal disease from 1960s to 1980s and in prevention and management of periodontal disease since the end of 1980s. This presentation will discuss about the prevalence of periodontal disease, treatment need, and provision of periodontal treatment in Korea, and how we could manage the periodontal disease efficiently in the future. According to an epidemiological study in Korea, periodontal disease(including gingivitis) was present in 82% of general population and periodontitis in 30-40% in adult population over 30y and juvenile periodontitis in 0.1% of adolescents. If we consider that at least 17% of these patients may have recurrent or refractory forms, there is obviously an abundance of disease that needs treatment, As a result of increase in life expectancy, senile population over 65 y will be increased from 6% in 1996 to 6.9% in 2000, and tooth retention rate and periodontal treatment need are expected to increase. Periodontists need all the help they can get from the general dentists to control periodontal disease. As for provision, postgraduate course in periodontics started in 1957 in Korea and produced over 700 specialized dentists in periodontics. One report indicated that the periodontists as well as general practitioners did periodontal therapy on only a few periodontal patients, because of specific control by current medical insurance system in Korea. Comprehensive periodontal examination is rarely done in local dental clinic. Therefore, enhancement of periodontal care in medical insurance system and education of simplified periodontal examination such as Periodontal Screening & Recording will make dentists diagnose and manage the management of adult patients is based on the recognition that there are multiple diseases, including gingivitis, chronic adlt periodontitis, and other more aggressive forms of periodontitis, and requires the earliest possible recognition of these three disease categories. In this presentation, we discuss practical approach using PSR to diagnose, manage and refer the patients, to facilitate the separation of the simple from the complex and the predictable from the unpredictable form of periodontal diseases and to integrate diagnostic and therapeutic techniques into private practice today.

• Journal of Periodontal and Implant Science
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• v.37 no.sup2
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• pp.339-351
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• 2007

Cytolethal distending toxin(CDT)은 세포 주기 중 G2에서 M 기로의 전환을 막아 세포의 증식을 억제할 수 있는 세균 단백 독소의 일종이다. 구강 미생물 중 유일하게 Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans)만이 이 CDT를 생성 할 수 있는 것으로 알려져 있다. A. actinomycetemcomitans는 localized aggressive periodontitis (LAP)의 원인균으로 여겨지며 비 운동성의 그람 음성 구간균이고 $37^{\circ}C$, 5% $CO_2$ 하에 성장이 왕성하다. A. actinomycetemcomitans의 CDT는 3개의 인접한 유전자인 cdtA, cdtB, cdtC에 의해 형성 되며 각각의 유전자에 대한 단백질의 기능은 아직 완전히 밝혀지지 않았다. 현재까지 연구에 의하면 cdtA는 CDT의 세포부착과 관련이 있는 것으로 여겨지며 이 유전자의 기능 이상 시 CDT의 독성 효과가 현저히 감소한다고 알려져 있다. 따라서 본 연구는 A. actinomycetemcomitans의 cdtA 유전자를 클로닝, 단백질 발현하여 향후 치주질환의 발병 과정에서 CdtA의 역할을 규명하고 질환의 예방 및 치료법에 도움을 주고자 하였다. A. actinomycetemcomitans Y4균주를 cdtA 유전자 클로닝을 위해 사용하였다. A. actinomycetemcomitans의 genomic DNA는 genomic DNA 추출 kit를 사용하여 분리하고 cdtA에 특이적인 primer를 이용하여 PCR을 통해 cdtA 유전자를 증폭하였다. 증폭된 cdtA 유전자를 T-vector에 클로닝 하였으며, 클로닝 된 cdtA 유전자는 단백질 발현을 위해 pRSET Avector에 서브클로닝 한 후 발현 균주인 BL21(DE3)를 이용하여 발현시켰다. 발현 후 Ni-NTA AP conjugate를 이용한 Western blot을 통해 pRSET-CDTA를 확인하였다.

• Journal of Periodontal and Implant Science
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• v.38 no.sup2
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• pp.335-342
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• 2008

Purpose: Cytolethal distending toxin (CDT) considered as a key factor of localized aggressive periodontitis, endocarditis, meningitis, and osteomyelitis is composed of five open reading frames (ORFs). Among of them, the individual role of CdtA and CdtC is not clear; several reports presents that CDT is an AB2 toxin and they enters the host cell via clathrin-coated pits or through the interaction with GM3 ganglioside. So, CdtA, CdtC, or both seem to be required for the delivery of the CdtB protein into the host cell. Moreover, recombinant CDT was suggested as good vaccine material and antibody against CDT can be used for neutralization or for a detection kit. Materials and Methods: We constructed the pET28a-cdtC plasmid from Aggregatibacter actinomycetemcomitans Y4 by genomic DNA PCR and expressed in BL21 (DE3) Escherichia coli system. We obtained the antibody against the recombinant CdtC in mice system. Using the anti-CdtC antibody, we test the native CdtC detection by ELISA and Western Blotting and confirm the expression time of native CdtC protein during the growth phase of A. actinomycetemcomitans. Results: In this study we reconstructed CdtC subunit of A. actinomycetemcomitans Y4 and generated the anti CdtC antibody against recombinant CdtC subunit expressed in E. coli system. Our anti CdtC antibody can be interacting with recombinant CdtC and native CDT in ELISA and Western system. Also, CDT holotoxin existed at 24h but not at 48h meaning that CDT holotoxin was assembled at specific time during the bacterial growth. Conclusion: In conclusion, we thought that our anti CdtC antibody could be used mucosal adjuvant or detection kit development, because it could interact with native CDT holotoxin.