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.

N-acetylcysteine prevents the development of gastritis induced by Helicobacter pylori infection

장성일,박은정,차정헌 한국미생물학회 2017 The journal of microbiology Vol.55 No.5

Helicobacter pylori (H. pylori) is a human gastric pathogen,causing various gastric diseases ranging from gastritis to gastricadenocarcinoma. It has been reported that combiningN-acetylcysteine (NAC) with conventional antibiotic therapyincreases the success rate of H. pylori eradication. We evaluatedthe effect of NAC itself on the growth and colonizationof H. pylori, and development of gastritis, using in vitroliquid culture system and in vivo animal models. H. pylorigrowth was evaluated in broth culture containing NAC. TheH. pylori load and histopathological scores of stomachs weremeasured in Mongolian gerbils infected with H. pylori strain7.13, and fed with NAC-containing diet. In liquid culture,NAC inhibited H. pylori growth in a concentration-dependentmanner. In the animal model, 3-day administration ofNAC after 1 week from infection reduced the H. pylori load;6-week administration of NAC after 1 week from infectionprevented the development of gastritis and reduced H. pyloricolonization. However, no reduction in the bacterial loador degree of gastritis was observed with a 6-week administrationof NAC following 6-week infection period. Our resultsindicate that NAC may exert a beneficial effect on reductionof bacterial colonization, and prevents the development ofsevere inflammation, in people with initial asymptomaticor mild H. pylori infection.

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

Cot kinase plays a critical role in Helicobacter pylori-induced IL-8 expression

장성일,김진문,차정헌 한국미생물학회 2017 The journal of microbiology Vol.55 No.4

Helicobacter pylori is a major pathogen causing various gastricdiseases including gastric cancer. Infection of H. pyloriinduces pro-inflammatory cytokine IL-8 expression in gastricepithelial cells in the initial inflammatory process. It has beenknown that H. pylori can modulate Ras-Raf-Mek-Erk signalpathway for IL-8 induction. Recently, it has been shown thatanother signal molecule, cancer Osaka thyroid oncogene/tumorprogression locus 2 (Cot/Tpl2) kinase, activates Mek andErk and plays a role in the Erk pathway, similar to MAP3Ksignal molecule Raf kinase. Therefore, the objective of thisstudy was to determine whether Cot kinase might be involvedin IL-8 induction caused by H. pylori infection. AGS gastricepithelial cells were infected by H. pylori strain G27 or its isogenicmutants lacking cagA or type IV secretion system followedby treatment with Cot kinase inhibitor (KI) or siRNAspecific for Cot kinase. Activation of Erk was assessed byWestern blot analysis and expression of IL-8 was measuredby ELISA. Treatment with Cot KI reduced both transient andsustained Erk activation. It also reduced early and late IL-8secretion in the gastric epithelial cell line. Furthermore, siRNAknockdown of Cot inhibited early and late IL-8 secretioninduced by H. pylori infection. Taken together, these resultssuggest that Cot kinase might play a critical role in H. pyloritype IV secretion apparatus-dependent early IL-8 secretionand CagA-dependent late IL-8 secretion as an alternativesignaling molecule in the Erk pathway.

Mouse Strain-Dependent Osteoclastogenesis in Response to Lipopolysaccharide

최호길,김진문,김봉주,유윤정,차정헌 한국미생물학회 2007 The journal of microbiology Vol.45 No.6

Bacterial lipopolysaccharide (LPS) is a potent stimulator of bone resorption in periodontitis. Co-culture systems of mouse calvaria-derived osteoblasts and bone marrow-derived preosteoclasts were used as an in vitro osteoclast differentiation. This study revealed that co-cultures using ddY or ICR mouse strain responded differently to LPS while responded equally to 1α,25(OH)2D3. Thus, the different response to LPS indicates dissimilarity of two mouse stains in their capacity for generating osteoclasts while the two mouse strains share the similarity in response to 1α,25(OH)2D3. To identify which cells between osteoblasts and preosteoclasts in the co-culture are responsible for the dissimilarity, the reciprocal co-cultures were performed between ddY and ICR mouse strains. The treatment of 1,25(OH)2D3 to ddY/ICR (osteoblasts from ddY/preosteoclasts from ICR) and ICR/ddY reciprocal co-cultures also showed the similarity. In case of LPS treatment, the results of ddY/ICR were similar to ddY/ddY and the results of the other reciprocal co-culture, ICR/ddY combination, were consistent with those of ICR/ICR. It suggests that the dissimilarity between the two mouse strains may resident in osteoblasts but not in preosteoclasts. Therefore, the osteoblast is responsible for mouse strain-dependent osteoclastogenesis in response to LPS. Although mouse models will continue to provide insights into molecular mechanisms of osteoclastogenesis, caution should be exercised when using different mouse strains, especially ddY and ICR strains as models for osteoclast differentiation.

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.

The influence of diabetes mellitus on periodontal tissues: a pilot study

엄유정,정의원,김창성,박은정,차정헌,유윤정,최성호 대한치주과학회 2010 Journal of Periodontal & Implant Science Vol.40 No.2

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 fi-bers. 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.

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.

Effect of globular adiponectin on interleukin-6 and interleukin-8 expression in periodontal ligament and gingival fibroblasts

박홍규,유윤정,박은정,김지혜,최성호,차정헌,Yang-Sin Lee 대한치주과학회 2011 Journal of Periodontal & Implant Science Vol.41 No.3

Purpose: Globular adiponectin (gAd) is a type of adipocytokine, which is mainly produced by adipose tissue. It has been reported that gAd acts as a pro- as well as an anti-inflammatory factor. Interleukin (IL)-6 and IL-8 are pro-inflammatory cytokines. To investigate the role of gAd on periodontal tissues, the expression of adiponectin receptor 1 (AdipoR1) and the effect of gAd on the expression of IL-6 and IL-8 were investigated in periodontal ligament (PDL) and gingival fibroblasts. Methods: PDL and gingival fibroblasts were cultured from human periodontal tissues. gAd derived from Escherichia coli and murine myeloma cells were used. The expression of AdipoR1 was estimated by reverse transcription-polymerase chain reaction and western blot. The expression of cytokines was measured by enzyme-linked immunosorbent assay. Results: PDL and gingival fibroblasts expressed both mRNA and protein of AdipoR1. gAd derived from E. coli increased the production of IL-6 and IL-8, but polymyxin B, an inhibitor of lipopolysaccharide (LPS), inhibited IL-6 and IL-8 production induced by gAd in both types of cells. gAd derived from murine myeloma cells did not induce IL-6 and IL-8 production in those cells. gAd derived from E. coli contained higher levels of LPS than gAd derived from murine myeloma cells. LPS increased production of IL-6 and IL-8 in PDL and gingival fibroblasts, but pretreatment of cells with gAd derived from murine myeloma cells did not inhibit LPS-induced IL-6 and IL-8 expression. Conclusions: Our results suggest that PDL and gingival fibroblasts express AdipoR1 and that gAd does not act as a modulator of IL-6 and IL-8 expression in PDL and gingival fibroblasts.