Full mouth disinfection therapy의 단기간 임상 효과 연구

조익현,정의원,차정헌,김중수,이대실,김창성,김종관,최성호,Cho, Ik-Hyun,Jung, Ui-Won,Cha, Jeong-Heon,Kim, Joong-Su,Lee, Dae-Sil,Kim, Chang-Seong,Kim, Chong-Kwan,Choi, Seong-Ho 대한치주과학회 2005 Journal of Periodontal & Implant Science Vol.35 No.3

The aim of this study is to determine whether full-mouth disinfection therapy(FMT) in our clinical setting would show better improvement of clinical parameters than partial mouth disinfection therapy(PMT) in chronic periodontitis and aggressive periodontitis patients. Among 12 patients, 6 were treated FMT and other 6 were treated PMT. Clinical parameters were calculated 3 months and 6 months after initial therapy. 1. There were no statistically significant differences between FMT and PMT in the reduction rate of bleeding on probing after 3 months, 6 months 2. Initial probing depth was 4-6mm, the mean probing depth after 3 months was 2.2mm vs 2.5mm(FMT vs PMT), after 6 months was 2.4mm vs 2.8mm. This was significantly lower in the FMT groups. 3. Initial probing depth was ${\geqq}$ 7mm, the reduction rate of mean probing depth during first 3 months was 4.8mm vs 4.1mm(FMT vs PMT), and 3 to 6 months was 0.5mm vs 0.3mm. This was significantly larger in the FMT groups. 4. Initial probing depth was 4-6mm, the mean clinical attachment level after 3 months was 2.3mm vs 2.7mm(FMT vs PMT), after 6 months was 2.7mm vs 3.0mm. This was significantly lower in the FMT groups. 5. Initial probing depth was ${\geqq}$ 7mm, the reduction rate of mean probing depth during first 3 months was 4.0mm vs 3.0mm(FMT vs PMT), and 3 to 6 months was 0mm vs -0.1mm. This was significantly larger in the FMT groups. Although the results provided us with succeccful clinical improvement in aggressive periodontitis, further research is needed to prove its additional benefit in the treatment of chronic periodontitis

Lipopolysaccharide가 파골세포 전구세포의 이동에 미치는 영향

이희영,이대실,차정헌,유윤정,Lee, Hee-Young,Lee, Dae-Sil,Cha, Jeong-Heon,Yoo, Yun-Jung 대한치주과학회 2007 Journal of Periodontal & Implant Science Vol.37 No.1

파골세포에 의한 골흡수는 1) 혈관을 통한 파골세포 전구세포의 골표면 이동 및 2) 골표면에서 파골세포 전구세포로부터 파골세포 분화 두 단계를 거쳐 일어난다. Stromal cell derived factor $(SDF)-1{\alpha}$ 는 파골세포 전구세포의 화학주성인자이며 matrix metalloproteinase (MMP)-9는 파골세포 전구세포의 이동에 관여하는 단백 분해효소이다. 파골세포 전구세포의 골표면 이동에 있어서 LPS의 역할을 규명하기 위하여 E. coli 및 Actinobacillus actinomycetecomitans LPS의 1) 파골세포 전구세포 유도능, 2) LPS에 의한 파골세포 전구세포의 이동에 있어서 MMP 및 $SDF-1{\alpha}$ 의 관련성을 평가하였다. LPS에 의한 차골세포 전구세포의 RAW 세포의 이동은 matrigel 또는 type I collagen을 도포한 transwell을 이용하여 평가하였으며 MMP-9 및 $SDF-1{\alpha}$ 의 발현은 RT=PCR 또는 ELISA로 평가하였다. 각 세균의 LPS는 matrigel 또는 type I collagen을 통한 파골세포 전구세포의 이동을 증가시켰다. MMP 억제제는 각 세균의 LPS에 의한 파골세포 전구세포의 이동을 억제하였다. LPS는 파골세포 전구세포의 MMP-9의 발현을 증가시켰다. 각 세균의 LPS는 마우스 두개골에서 분리한 조골세포의 $SDF-1{\alpha}$ 의 발현을 증가시켰다. $SDF-1{\alpha}$ 을 함유한 LPS 처리 조골세포 배양상층액은 파골세포 전구세포의 이동을 증가시켰으며 anti $SDF-1{\alpha}$ Ab는 LPS처리 세포 배양상층액에 의한 파골세포 전구세포의 이동을 억제하였다. 이들 결과는 LPS가 파골세포 전구세포에서는 MMP-9을 조골세포에서는 $SDF-1{\alpha}$ 의 발현을 증가시켜 파골세포 전구세포의 이동을 촉진 시킬 수 있음을 시사한다.

치주인대세포에서 Aggregatibacter actinomycetemcomitans의 IL-8 및 활성산소종 유도능

이양신,박홍규,김성환,차정헌,유윤정,Lee, Yang-Sin,Park, Hong-Gyu,Kim, Sung-Whan,Cha, Jeong-Heon,Yoo, Yun-Jung 대한치주과학회 2009 Journal of Periodontal & Implant Science Vol.39 No.3

Purpose: Interleukin (IL)-8 is one of pro-inflammatory cytokines. Reactive oxygen species (ROS) are reduced metabolites of $O_2$. Aggregatibacter actinomycetemcomitans is one of representative periodontopathogens. To investigate the role of A. actinomycetemcomitans in IL-8 expression of periodontal ligament (PDL) cells, we estimated the production of IL-8 and ROS in A. actinomycetemcomitans treated PDL cells. Methods: The IL-8 production was determined by enzyme-linked immunosorbent assay. The ROS production was estimated using H2DCFDA and FACS. Results: A. actinomycetemcomitans increased the production of IL-8 and ROS at 10, 100, and 500 multiplicity of infection. N-acetylcysteine, an antioxidant of ROS, down-regulated the production of IL-8 induced by A. actinomycetemcomitans. Conclusions: These results suggest that A. actinomycetemcomitans induces IL-8 production and ROS may act as a mediator in this process.

조골세포에서 Aggregatibacter actinomycetemcomitans 생균의 파골세포분화유도 cytokine 발현 유도능 및 침투능

최호길,이양신,김민영,김경대,차정헌,유윤정,Choi, Ho-Kil,Lee, Yang-Sin,Kim, Min-Young,Kim, Kyoung-Dae,Cha, Jeong-Heon,Yoo, Yun-Jung 대한치주과학회 2007 Journal of Periodontal & Implant Science Vol.37 No.3

Osteoblasts regulate osteoclastogenesis by production of various cytokines. Aggregatibacter(A) ac-tinomycetemcomitans is one of periodontopathogens which invades gingival tissue. Therefore, clarifying the effect of alive A. actinomycetemcomitans on osteoblasts is important to understand the mechanism of alveolar bone resorption in periodontitis. We investigated induction of osteoclastogenesis-inducing cytokines, adherence, and invasion by A. actinomycetemcomitans in osteoblasts. Osteoblasts were isolated from mouse calvaria and expression of cytokines was determined by RT-PCR. When the ratio of the number of A. actinomycetemcomtians to the number of osteoblasts was 10:1, 50:1 and 100:1, RANKL mRNA expression was increased. A. actinomycetemcomitans also increased expression of macrophage inflammatory protein (MIP) -1${\alpha}$, interleukin (IL)-1${\beta}$, and tumor necrosis factor (TNF)-${\alpha}$. A. actinomycetemcomitans attached to and invaded osteoblasts at ratio of 1000:1. These results suggest that A. actinomycetemcomitans increases osteoclastogenesis-inducing ability of osteoblasts by stimulating the expression of RANKL, MIP-1${\alpha}$,IL-1${\beta}$, and TNF-${\alpha}$ and that invasion of A. actinomycetemcomitans provides a means by which the bacteria escape from immune system and antibiotic therapy.

Treponema lecithinolyticum lipopolysaccharide에 의한 matrix metalloproteinase-9의 발현

남정아,문선영,이진욱,차정헌,최봉규,유윤정,Nam, Jeong-Ah,Moon, Sun-Young,Lee, Jin-Wook,Cha, Jeong-Heon,Choi, Bong-Kyu,Yoo, Yun-Jung 대한치주과학회 2005 Journal of Periodontal & Implant Science Vol.35 No.3

Bone resorption involves sequential stages of osteoclast precursor migration and differentiation of osteoclast precursors into multinucleated osteoclasts. Stromal cell derived factor (SDF)-1 is a chemotactic factor for osteoclast precursor migration. Matrix metalloproteinase (MMP)-9 is involved in migration of osteoclast precursors and activation of $interleukin(IL)-1{\beta}$. Alveolar bone destruction is a characteristic feature of periodontal disease. Treponema lecithinolyticum is a oral spirochete isolated from the periodontal lesions. The effect of lipopolysaccharide(LPS) from T. lecithinolyticum on expression of SDF-1 and MMP-9 was examined in cocultures of bone marrow cells and osteblasts derived from mouse calvariae. T. lecithinolyticum LPS increased expression of MMP-9 in the coculture. Polymyxin B, an inhibitor of LPS, abolished the increase of MMP-9 mRNA expression by LPS. LPS did not increase the expression of SDF-1, $IL-1{\beta}$ and tumor necrosis $factor(TNF)-{\alpha}$ mRNA in cocultures. Prostaglandin $E_2(PGE_2)$ up-regulated the expression of MMP-9 and NS398, an inhibitor of $PGE_2$ synthesis, down-regulated the induction of MMP-9 expression by T. lecitbinolyticm LPS. These results suggest that T. lecitbinolyticm LPS increases MMP-9 expression in bone cells via $PGE_2$ and that the induction of MMP-9 expression by T. lecitbinolyticm LPS is involved in alveolar bone destruction of periodontitis patients by the increase of osteoclast precursor migration and the activation of bone resorption-inducing cytokine.

CagL 재조합 단백질 접종후에 Mongolian gerbil에서 나타나는 Helicobacter pylori 감염에 대한 반응

박은정,장성일,최윤희,김진문,김애련,김지혜,우계형,유윤정,이성행,차정헌,Bak, Eun-Jung,Jang, Sung-Il,Choi, Yun-Hui,Kim, Jin-Moon,Kim, Ae-Ryun,Kim, Ji-Hye,Woo, Gye-Hyeong,Yoo, Yun-Jung,Lee, Sung-Haeng,Cha, Jeong-Heon 한국미생물학회 2012 미생물학회지 Vol.48 No.2

Helicobacter pylori는 만성 위염, 소화성 궤양, 위암의 중요한 역학적 인자중 하나이다. H. pylori의 독성인자중 CagL은 숙주 세포와 H. pylori의 제 4형 분비기관(Type 4 secretion system)을 연결하는 adhesin으로 작용하는 섬모 단백질로 H. pylori가 발병하는데 중요한 역할을 하는 것으로 알려져 있다. 이번 연구는 저빌에 H. pylori를 감염시킨 동물 모델을 이용하여 CagL 재조합 단백질을 면역화시켰을 때 나타나는 효과를 평가하였다. 재조합 CagL은 클론되었고, 과발현시켜 정제하여 준비하였다 저빌은 H. pylori 감염 대조군과 H. pylori 감염 CagL 재조합 단백질 접종군으로 분류하였고, 접종시 알루미늄 애쥬번트를 사용하였다. 일주일 간격으로 4회 근육내 접종하였고, 마지막 접종 일주일 후, 모든 저빌에 H. pylori 7.13 균주를 $1{\times}10^9\;bacteria/500{\mu}l$ 농도로 위내 투여하였다. H. pylori 감염 6주째 모든 저빌을 희생하여 혈청 IgG 반응평가를 위한 ELISA를 실시하였고, 위에서는 집락화된 H. pylori의 수평가, 병리조직학적 평가 및 사이토카인 유전자발현을 조사하였다. CagL 재조합 단백질접종 일주일 후부터 H. pylori 감염 CagL 재조합 단백질 접종군의 혈청내 IgG 항체형성이 유의적으로 증가하였다. 위에서의 집락화된 세균수는 두군의 차이가 없었다. 저빌 체중에 대한 위무게 비율는 H. pylori 감염 CagL 재조합 단백질 접종군이 유의적으로 감소하였으나 병리조직학적 평가에서는 유의적인 차이는 확인하지 못하였다. 위에서의 IL-$1{\beta}$와 KC (IL-8 homologues)의 유전자발현 정도도 두 군사이에 유의적인 차이는 없었다. 이번 결과는 CagL 재조합 단백질의 접종은 IgG 항체형성은 효과적으로 자극하였지만 면역화된 숙주에서 세균 집락화의 감소 및 병변형성의 방어까지는 유도하지 못한 것으로 나타났으며, 앞으로 H. pylori 감염에 대해 유효한 면역 반응 및 질병 방어 효과를 나타내기 위해서 CagL을 포함한 다른 종류의 재조합 항원 사용 및 보조적으로 전신 면역 및 점막 면역을 효과적으로 유도하기 위해 안정성있는 애쥬번트의 사용을 고려해야 할 것으로 사료된다. Helicobacter pylori is an important factor of chronic gastritis, digestive ulcer, and stomach cancer. CagL, a virulence factor of H. pylori, is well-known as a pilus protein which acts as adhesion to host cell and a component of Type 4 secretion system. In this study, we evaluated the protective response of recombinant CagL protein (rCagL) using Mongolian gerbil animal model for H. pylori infection. The cagL gene was cloned from 26695 H. pylori followed by over-expression and purification of the protein in E. coli. Mongolian gerbils were immunized with rCagL protein mixed with aluminum adjuvant via intramuscular injections once a week during 4 weeks. At a week after the last immunization, the Mongolian gerbils were administrated with H. pylori 7.13 strain into the stomach and sacrificed to measure antibody titer on rCagL by ELISA and bacterial colonization in the stomach, and to examine the histopathological changes and cytokine expression at 6 week after challenge. Antibody titers on recombinant protein were significantly increased from a week after the first immunization. There was no significant change of the number of bacterial colony between control group and immunized group. The relative stomach weight was significantly decreased in immunized group, but the significant change of histopathological assessment was not observed in the stomach. Cytokine expression such as IL-$1{\beta}$ and KC also was not significantly different between control and immunized groups. These results indicate that rCagL could effectively induce the formation of the specific IgG antibodies. However, bacterial colonization and histopathological lesions could not be inhibited by the immunization in the stomach, indicating not enough protection against H. pylori infection. We consider that along with CagL other adequate antigens could be needed stimulating immune response and inducing protective effects against gastric disease, and also a better adjuvant could be considered.

치주염 유발 쥐에서 상아질파괴세포와 뼈파괴세포의 형성

이동은 ( Dong Eun Lee ),김지혜 ( Ji Hye Kim ),신동하 ( Dong Ha Shin ),차정헌 ( Jeong Heon Cha ),박은정 ( Eun Jung Bak ),유윤정 ( Yun Jung Yoo ) 한국치위생과학회 2015 치위생과학회지 Vol.15 No.3

Odontoclasts and osteoclasts resorb tooth root and alveolar bone, respectively. Many studies have focused on osteoclast formation in periodontitis, but effect of periodontitis on odontoclast formation is not clearly clarified. In this study, we observed formation of odontoclasts as well as osteoclasts in rats with ligature-induced periodontitis. To induce periodontitis, ligatures were placed around the first molars in left mandibles of rats. Rats were sacrificed at days 1, 3, and 10 after ligation. After tartrate resistant acid phosphatase (TRAP) staining in mandible section, the number of TRAP-positive odontoclasts and osteoclasts were histologically counted along the root and the alveolar bone surfaces of tooth, respectively. Odontoclasts increased until day 10 in mesial and furcation root surface, but did not increase in distal root surface. When compared odontoclast formation to osteoclast formation in mesial surface, osteoclasts peaked at day 3, and then decreased gradually, whereas odontoclasts were continuously increased until day 10. The number of odontoclasts was lower than that of osteoclasts before and after periodontitis induction. These indicate that periodontitis increased formation of odontoclasts as well as osteoclasts, but odontoclast formation occurs slower and weaker than that of osteoclasts.

휴대용 플라즈마 소자를 이용한 액체 내 박테리아 살균 연구

이힘찬(Him-Chan Lee),임토우(To-Woo Lim),서지원(Ji-Won Seo),김영민(Youngmin Kim),차정헌(Jeong-Heon Cha) 대한전기학회 2022 대한전기학회 학술대회 논문집 Vol.2022 No.7