녹용 추출물에 의한 MC3T3세포의 분화 촉진

유윤정,이현정,임소형,강정화,이은희,옥승호,최봉규,전길자,Yoo, Yun-Jung,Lee, Hyun-Jung,Lim, So-hyung,Kang, Jung-Hwa,Lee, Eun-Hui,Ohk, Seung-Ho,Choi, Bong-Kyu,Jhon, Gil-Ja 대한치주과학회 2000 Journal of Periodontal & Implant Science Vol.30 No.4

Deer antler has been widely prescribed in Chinese and Korean pharmacology. Although there have been several reports concerning the effects of deer antler, such as anti-aging action, anti-inflammatory activity, antifungal action and regulatory activity of the level of glucose, the effect on bone has not determined yet. The purpose of this study was to examine the effect of deer antler on osteoblast differentiation. Hexane extract(CN-H) and chloroform extract(CN-C) were acquired from deer antler(Cervus nippon) and MC3T3-E1 pre-osteoblasts were cultured in the presence or absence of each extract. Osteoblast differentiation was estimated with the formation of mineralized nodules and the mRNA expression of alkaline phosphatase(ALP), osteocalcin(OC) and bone sialoprotein(BSP) which are markers of osteoblast differentiation. Non-treated group did not show mineralized nodule. CN-C or CN-H-treated group showed minerlaized nodules in 16 days. In northern blot analysis, CN-C or CN-H-treated group showed the elevated expression of ALP, BSP and OC in 16 days. These results suggest the possibility to develop deer antler as a bone regenerative agent in periodontal therapy by showing the stimulating activity of deer antler on differentiation of osteoblast.

자몽종자추출물과 자일리톨이 배합된 껌의 치은염 예방 및 항균효과

진미성,유윤정,최봉규,이희영,김미정,노회진,박종섭,조규성,김종관,최성호,Jin, Mi-Sung,Yoo, Yun-Jung,Choi, Bong-Kyu,Lee, Hee-Young,Kim, Mi-Jung,Roh, Hoe-Jin,Park, Jong-Sub,Cho, Kyoo-Sung,Kim, Chong-Kwan,Choi, Seong-Ho 대한치주과학회 2003 Journal of Periodontal & Implant Science Vol.33 No.3

Grapefruit seed extract has been reported to have antimicrobial effect. The purpose of this study was to evaluate the antimicrobial and anti-gingivitis effect of chewing gum containing grapefruit seed extract and xylitol. 40 healthy subjects with gingivitis or early periodontitis were divided into two groups. Subjects in the experimental group chewed gum containing grapefruit seed extract and xylitol while subjects in the control group chewed gum containing only xylitol. All subjects received scaling and tooth brushing instruction. 1 week after scaling was set as baseline. Gingival index and plaque index were scored at baseline, 1 week, 2 week, 3 week and 4 week. Bleeding index, probing pocket depth and clinical attachment level were scored at baseline, 2 week and 4 week. The number of total bacteria and Streptococcus mutans in unstimulated saliva of experimental group were counted at 1 week, 2 week, 3 week and 4 week. Gingival indices of experimental group and control group at baseline, 1 week, 2 week, 3 week and 4 week were 0.850${\pm}$0.298, 0.575${\pm}$0.345, 0.533${\pm}$0.332, 0.459${\pm}$0.311, 0.408${\pm}$0.224 and 0.758${\pm}$0.379, 0.592${\pm}$0.276, 0.563${\pm}$0.281, 0.454${\pm}$0.194, 0.413${\pm}$0.199 (mean${\pm}$SD), respectively. Plaque indices of experimental group and control group at baseline, 1 week. 2 week, 3 week and 4 week were 0.497${\pm}$0.500, 0.375${\pm}$0.484, 0.332${\pm}$0.471, 0,286${\pm}$0.452, 0.210${\pm}$0.407 and 0.411${\pm}$0.492, 0.375${\pm}$0.484, 0.354${\pm}$0.479, 0.313${\pm}$0.463, 0.193${\pm}$0.395, respectively. Bleeding indices of experimental group and control group at baseline, 2 week and 4 week were 0.377${\pm}$0.177, 0.298${\pm}$0.152, 0.192${\pm}$0.108 and 0.383${\pm}$0.124, 0.318${\pm}$0.153, 0.225${\pm}$0.126, respectively. Probing pocket depth of experimental group and control group at baseline, 2 week and 4 week were 2.56${\pm}$1.00, 2.40${\pm}$0.65, 2.23${\pm}$0.64 and 2.45${\pm}$0.682.37${\pm}$0.57, 2.19${\pm}$0.57, respectively. Clinical attachment level of experimental group and control group at baseline, 2 week and 4 week were 2.58${\pm}$1.01, 2.43${\pm}$0.67, 2.26${\pm}$0.65 and 2.49${\pm}$0.70, 2.40${\pm}$0.59, 2.22${\pm}$0.62, respectively. The % of reduction of total bacteria in saliva of experimental group at 2 week, 3 week and 4 week were 46 ${\pm}$ 53%, 53 ${\pm}$ 5% and 69 ${\pm}$ 33%. The % of reduction of Streptococcus mutans count in saliva of experimental group at 2 week, 3 week and 4 week were 52${\pm}$69%, 88${\pm}$30% and 89${\pm}$17%. From these findings, it can be concluded that regular use of grapefruit seed extract /xylitol chewing gum may be effective to control and prevent gingivitis and may have caries-preventive effect.

Cetylpyridinium Chloride(CPC), NaF 및 Ursodeoxycholic acid(UDCA) 혼합물의 주요 치주병원균에 대한 in Vitro 항균효과

김종관,최봉규,유윤정,김상년,석재균,김문무,Kim, Chong-Kwan,Choi, Bong-Kyu,Yoo, Yun-Jung,Kim, Sang-Nyun,Seok, Jae-Kyun,Kim, Moon-Moo 대한치주과학회 1999 Journal of Periodontal & Implant Science Vol.29 No.2

The antibacterial efficacy of a mouthrinse(Denta Gargle) containing CPC(cetylpyridinium chloride), NaF and UDCA(ursodeoxycholic acid), on major periodontopathogens, was in vitro examined and compared with that of Listerine by a broth dilution method. The bacteria tested were Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Fusobacterium nucleatum subsp. vincentii, Prevotella intermedia, Porphyromonas gingivalis and Treponema denticola. The growth of all the bacteria were completely inhibited by a 1-min exposure to the both mouthrinses. When diluted at 1:5 or more, all bacteria analyzed but P. intermedia were not inhibited by Listerine. In contrast, Denta Gargle showed highly increased maximum inhibitory dilutions(MID) against all periodontopathogens included in this study, with MIDs ranging from 5-fold(F. nucleatum) to 160-fold dilutions(P. intermedia). The MIDs against A. actinomycetemcomitans, B. forsythus, P. gingivalis and T. denticola. were 1:40, 1:80, 1:80 and 1:80, respectively.

홍화씨 분획 추출물이 치주인대 섬유아세포와 MC3T3-E1 세포에 미치는 영향

허지선,강정화,유윤정,김창성,조규성,최성호,Huh, Ji-Sun,Kang, Jung-Hwa,Yoo, Yun-Jung,Kim, Chang-Sung,Cho, Kyoo-Sung,Choi, Seong-Ho 대한치주과학회 2001 Journal of Periodontal & Implant Science Vol.31 No.4

Recently, use of natural medicine is getting more attention, and some of them are believed to be effective in the treatment of periodontitis. Among them, the seeds of safflower(Carthamus tinctrorius L.) have been proven to be effective through its use in bone diseases such as fracture and osteoporosis. During the last few years, studies using the seeds of safflower gown in Korea have been active, and it has been reported that safflower seed extract increase the proliferation and the alkaline phosphatase(ALP) activity of human periodontal ligament fibroblast(hPDLF), osteoblast, and that they promote the mineralization process. In animal studies, when safflower seed extract were administered orally new bone formation was promoted. Recently, in an effort to find out the most effective osteogenic components, among many components of the safflower seed, various safflower seed fraction extracts were obtained by multistep extraction of the safflower components using various solvents. Among these, saf-M-W fraction extracted by methanol and water was most effective in increasing osteogenic potential of osteoblasts. In this study, the effect of safflower seed fraction extract, saf-M-W, on the growth and differentiation of hPDLF and MC3T3-E1 cell was investigated. The toxicity of saf-M-W on both cells was measured using M'IT(3-(4,5dimethylthiazol-2-y1)-2,5-diphenyl tetrazolium bromide) test, and ALP activity was measured using the colorimetric assay of hPDLF. In addition, in MC3T3-El cells, the expression of ALP, bone sialoprotein(BSP) mRNA was observed using Northern blot, and the mineralized nodule formation Was observed using von Kossa stain and phase-contrast microscope. 1. In concentrations below $10{\mu}g/ml$, saf-M-W didn't show any toxicity on hPDLF and MC3T3-El cell. 2. The change in saf-M-W concentration had no effect on the ALP activity of hPDLF. 3. In MC3T-E1 cells, mRNA expressions of ALP and BSP were greater in the experimental group treated with $10{\mu}g/ml$ concentration of saf-M-W compared with the control group. 4. In MC3T3-El cells, abundance of mineralized nodules were formed in the experimental group treated with $10{\mu}g/ml$ Concentration of saf-M-W, while no mineralized nodule was formed in the control group. These results suggest that safflower seed fraction extract, saf-M-W. didn't show any toxicity on hPDLF and MC3T3-E1 cell at concentrations below $10{\mu}g/ml$ and effectively enhanced the differentiation and osteogenic potential of MC3T3-El cell.

키토산이 치주인대 섬유아세포에 미치는 영향

백정원,이현정,유윤정,조규성,김종관,최성호,Paik, Jeong-Won,Lee, Hyun-jung,Yoo, Yun-Jung,Cho, Kyoo-Sung,Kim, Chong-Kwan,Choi, Seong-Ho 대한치주과학회 2001 Journal of Periodontal & Implant Science Vol.31 No.4

Periodontal therapy has dealt primarily with attempts at arresting progression of disease, however, more recent techniques have focused on regenerating the periodontal ligament having the capacity to regenerate the periodontium. The effect of chitosan(poly-N-acetyl glucosaminoglycan), a carbohydrate biopolymer extracted from chitin, on periodontal ligament regeneration is of particular interest. The purpose of this study was to evaluate the effect of chitosan on the human periodontal ligament fibroblasts(hPDLFs) in vitro, with special focus on their proliferative properties by M'IT assay, the synthesis of type I collagen by reverse transcription-polymerase chain reaction(RT-PCR) and the activity of alkaline phosphatase(ALP). Fibroblast populations were obtained from individuals with a healthy periodontium and cultured with ${\alpha}MEM$ as the control group. The experimental groups were cultured with chitosan in concentration of 0.01,0.1, 1,2mg/ml. The results are as follows; 1. Chitosan-induced proliferative responses of hPDLFs reached a plateau at the concentration of O.lmg/ml(p<0.05). 2. When hPDLFs were stimulated with 0.lmg/ml chitosan, mRNA expression of type I collagen was up-regulated. 3. When hPDLFs were stimulated with 0.lmg/ml chitosan, ALP activity was significantly up-regulated(p<0.05). In summary, chitosan(0.lmg/ml) enhanced the type I collagen synthesis in the early stage, and afterwards, facilitated differentiation into osteogenic cells. The results of this in vitro experiment suggest that chitosan potentiates the differentiation of osteoprogenitor cells and may facilitate the formation of bone.

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.

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.

($17{\beta}$-Estradiol 및 1,25-Dihydroxyvitamin $D_3$가 치주인대 세포의 Interleukin-6의 생성에 미치는 영향

곽월아,최봉규,이현정,유윤정,Kwak, Wall-Ah,Choi, Bong-Kyu,Lee, Hyun-Jung,Yoo, Yun-Jung 대한치주과학회 1999 Journal of Periodontal & Implant Science Vol.29 No.3

Interleukin-6(IL-6) stimulate osteoclast differentiation. $17{\beta}$-estradiol, 1,25-dihydroxyvitamin $D_3$(1,25-$(OH)_2D_3$) and interleukin-$1{\beta}$ inhibit or stimulate osteoclast differentiation by decreasing or increasing the synthesis of interleukin-6(IL-6) from stromal/osteoblastic cells, respectively. Periodontal ligament(PDL) cells reside between the alveolar bone and the cementum and have osteoblastic characteristics. To estimate the effect of $17{\beta}$-estradiol and 1,25$(OH)_2D_3$ on IL-6 production of PDL cells, PDL cells were treated with $17{\beta}$-estradiol or 1,25-$(OH)_2D_3$ in the absence or the presence of IL-$1{\beta}$. The concentration of IL-6 produced form PDL cells was determined by enzym linked immunosorbent assay(ELISA). In unstimulated PDL cells, we detected constitutive production of IL-6 at 1st and 2nd day. IL-$1{\beta}$ increased IL-6 synthesis at 1st day and 2nd day. $17{\beta}$-estradiol had no significant effect on the secretion of this cytokine, either constitutively or after stimulation with IL- $1{\beta}$(0.05 ng/ml). 1,25-$(OH)_2D_3$($10^{-8}M$) decreased not only constitutive IL-6 production but also IL-$1{\beta}$-induced IL-6 production at 2nd day. These results suggest that 1,25-$(OH)_2D_3$ may control IL-$1{\beta}$-induced osteoclast differentiation by decreasing IL-$1{\beta}$-induced IL-6 secretion of PDL cells.

속단의 dichloromethane 분획물이 마우스 두개골 세포의 분화에 미치는 영향

김동진,윤정호,정의원,유윤정,김윤철,유형근,김종관,최성호,Kim, Dong-Jin,Yun, Jeong-Ho,Jung, Ui-Won,Yoo, Yun-Jung,Kim, Yun-Chul,You, Hyung-Keun,Kim, Chong-Kwan,Choi, Sung-Ho 대한치주과학회 2004 Journal of Periodontal & Implant Science Vol.34 No.4

The purpose of this study was to evaluate the effects of DFPR on differentiation of mouse calvarial cell in vitro, to examine the possibility for periodontal regeneration. $10{\mu}g/ml$ of DFPR was used as experimental concentration. osteogenic medium only was assigned as control, Experimental 1 was supplemented with 10nM dexamethasone, Experimental 2 with $10{\mu}g/ml$ DFPR and Experimental 3 with l0nM dexamethasone + $10{\mu}g/ml$ DFPR. cellular activity was evaluated by MTT method at 8, 12, 16 days, expression of mRNA of ALP, osteopontin, osteocalcin, collagen type-l was detected by RT-PCR method at 4, 8, 12, 16 days of culture. extent of mineralization was observed by Von Kossa staining at 16 day of culture. The results are as follows 1)Any acceleration of differentiation was not observed at expression of differentiation marker, 2) Decrease in expression of extracelluar matrix and in bone nodule formation was observed The results suggested that DFPR have negative effect on the rate of differentiation on rat calvarial cell, decrease extracelluar matrix formation ,decrease bone nodule formation. Ongoing studies are necessary in order to determine effect of DFPR on periodontal regeneration.

Treponema lecithinolyticum 주외막단백질 (MspTL)의 역할 규명

김종관(Chong-Kwan Kim),유윤정(Yun-Jung Yoo),김영호(Young-Ho Kim),최봉규(Bong-Kyu Choi) 대한미생물학회 2002 Journal of Bacteriology and Virology Vol.32 No.1