황련과 Centella asiatica 추출물이 치은 섬유모세포에 미치는 영향

유형근,You, Hyung-Keun 대한치주과학회 1996 Journal of Periodontal & Implant Science Vol.26 No.3

Periodontal regeneration requires the migration and proliferation of gingival fibroblasts and periodontal ligament cells. These cellular events are influenced and regulated by growth factors and some drugs. The purpose of this study is to examine the effect of Rhizoma coptidis and Centella asiatica extracts on human gingival fibroblasts. Gingival fibroblasts were primarily cultured from extracted premolar with non-periodontal diseases. Cells were cultured with ${\alpha}-MEM$ at $37^{\circ}C$, 5% $CO_2$, 100% humidity incubator for 2 or 3 days, as a measure of cell proliferation potential, it was examined that the DNA synthesis using $[^3H]-thyrnidine$ incorporation, the cell numbers (with or without dye), and cell viabilities. Rhizoma coptidis is increased the proliferation of gingival fibroblasts at concentration of $10^{-9}g/ml$, but Centella asiatica is decreased the proliferation at all concentrations. This study demonstrated that Rhizoma coptidis is a potential mitogen for human gingival fibroblasts in vitro, and we can expect the usefulness of this drug in periodontal regeneration.

Effect of Mechanical Stress on the Proliferation and Expression of Cell Cycle Regulators in Human Periodontal Ligament Cells

유형근,신형식,이진,민병무,You, Hyung-Keun,Shin, Hyung-Shik,Lee, Gene,Min, Byung-Moo The Korean Academy of Periodontoloy 1999 Journal of Periodontal & Implant Science Vol.29 No.3

치주인대세포는 치주인대의 유지와 개조에 있어서 중요한 역할을 담당하는 섬유아세포성 세포로서, 세포에 가해진 여러가지 조건에 따라 다양한 표현형의 변화를 나타내는 것으로 알려져 있다. 기계적 응력은 치주인대세포의 세포증 식과 밀접히 연관되어 있는 것으로 알려져 있으며, 이는 세포주기 조절인자들의 발현을 증가 시킴으로써 이루어질 것으로 생각되나 그 자세 한 작용기전은 알려져 있지 않다. 그러므로 이 연구의 목적은 기계적 응력이 사람 치주인대세 포의 세포증식과 세포주기 조절인자의 발현에 미치는 영향을 연구하기 위하여 사람 치주인대 세포에 기계적 응력을 가한 후 세포증식을 관찰하고 , 세포주기조절인자들인 p 53 , $p21^{WAF1/CIP1}$ cyclin-dependent kinases(cdks), cyclins 및 proliferating cell nuclear antigen(PCNA)의 단백질 발현 변화를 연구하였다. 본 연구에 사용한 사람 치주인 대세포는 교정치료를 목적으로 발거한 건전한 사람 소구치의 치주인대로부터 explantation culture하여 얻은 후 계대배양을 시행하여 제6 계대의 세포를 사용하였다. 배양한 사람 치주인 대세포를 55-mm Petriperm dish당 $1{\times}10^4$ 개를 분주하고, dish당 1kg의 기계적 응력을 가하면서 12일동안 세포배양을 시행하였다. 사람 치주인대세포의 세포증식은 기계적 응력을 가한 후 8-12일 사이에 현저히 증가하였으며, PCNA 단백질의 발현은 기계적 응력을 가한 후 6-10일 사이에 현저히 증가하였다. 또한 기계적 응력은 사람 치주 대세포의 cdk4, cdk6, cdk2 및 cyclin D1 단백질의 발현을 다소 증가 시켰으나, p53 및 $p21^{WAF1/CIP1}$ 단백질의 발현은 큰 변화가 없었다. 이상의 결과 서 기계적 응력은 사람 치주인대세포 의 p53 및 $p21^{WAF1/CIP1}$ 단백질 발현의 변화 없이 cdks 단백질 발현을 증가시킴으로써 세포증식을 증가시키는 것으로 생각된다.

엽산의 구강세정이 치은염증에 대해 미치는 효과에 관한 연구

유형근,김창원,신형식,You, Hyung-Keun,Kim, Chang-Won,Shin, Hyung-Shik 대한치주과학회 1990 Journal of Periodontal & Implant Science Vol.20 No.2

한약재 물 추출물의 Monoamine Oxidase B의 활성 억제 및 항산화 활성 검색

이형철(Hyung Chul Lee),황상구(Sang-Gu Hwang),김대근(Dae Gun Kim),주성민(Sung Min Ju),유형근(Hyung-Keun You),김원신(Won Shin Kim),안원근(Won Gun An),전병훈(Byung Hun Jeon) 한의병리학회 2002 동의생리병리학회지 Vol.16 No.3

Minocycline 및 $TGF-{\beta}1$이 배양 인체 치은섬유모세포와 치주인대세포에 미치는 영향

윤동환,유형근,신형식,Yoon, Dong-Hwan,You, Hyung-Keun,Shin, Hyung-Shik 대한치주과학회 1996 Journal of Periodontal & Implant Science Vol.26 No.1

One of the initial events required for periodontal regeneration is the attachment, spreading and proliferation of fibroblasts at the healing sites. These have been reported that minocycline stimulates the attachment of gingival fibroblasts and periodontal ligament cells and $TGF-{\beta}1$ enhances the proliferation of periodontal ligament cells. The purpose of this study was to evaluate and confirm the effect of minocycline and $TGF-{\beta}1$ on human gingival fibroblasts and periodontal ligament cells. That gingival fibroblasts and periodontal ligament cells used in this study were obtained from the explants of healthy periodontal ligaments and gingival tissues of extracted 3rd molars or premolar teeth extracted from the patients with orthodontic treatment. The cells were cultured in ${\alpha}-MEM$(minimal essential medium) supplemented with antibiotics and FBS(fetal bovine serum) at $37^{\circ}C$ in a humidified atmosphere of 5% carbon dioxide-95% air. Cells were used between the 5th to 8th passage in this study. The attachment and activity of both cells were evaluated by MTT assay. The results were as follows: 1. Maximum gingival fibroblast attachment was seen at a $50{\mu}g/ml$ dose of minocycline, while maximum periodontal ligament cell attachment was seen at a $100{\mu}g/ml$, and exposure of both cells to minocycline above maximal attachment dose results in a decline from maximum attachment. 2. The activity values of both cells tested minocycline were below to the control activity values at all concentrations. 3. The attachment values of both cells tested $TGF-{\beta}1$ were below or similar to control attachment values. On the above the findings, minocycline stimulated the cell attachment of gingival fibroblasts and periodontal ligament cells and $TGF-{\beta}1$ enhances the cell activity of periodontal ligament cells.

($TGF-{\beta}$)이 Minocycline을 전처리한 사람 치주인대세포의 활성에 미치는 영향

양승오,유형근,신형식,Yang, Seung-Oh,You, Hyung-Keun,Shin, Hyung-Shik 대한치주과학회 1996 Journal of Periodontal & Implant Science Vol.26 No.2

The initial events required for periodontal regeneration is the attachment, spreading, and proliferation of appropriated cells at the healing sites. These have been reported that minocycline stimulates the attachment of periodontal ligament cells, and also $TGF-{\beta}1$ enhances the proliferation of periodontal ligament cells. The purpose of the present study was to evaluate the effects of $TGF-{\beta}1$ on the cellular activity of minocycline treated human periodontal ligament cells. Periodontal ligament cells were obtained from the explants of healthy periodontal ligaments of extracted 3rd molars or premolar teeth extracted from the patients for orthodontic treatment. The cells were cultured in minimal essential medium(${\alpha}-MEM$) supplemented with 10.000units/ml penicillin, $10,000{\mu}g/ml$ streptomycin and 10% FBS(fetal bovine serum) at $37^{\circ}C$ in a humidified atmosphere of 5% carbon dioxide and the 5th to the 8th passages of the cells were used. To evaluate the effect of minocycline on cell attachment, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After trypsinization, the cells were counted with hemocytometer and were taken photographs for observation of cellular morphology. To evaluate the effect of $TGF-{\beta}1$ on minocycline-pretreated periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4$ cells/ well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After incubation, 1 and 10ng/ml of $rh-TGF-{\beta}1$ were also added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay, DNA synthesis($^3H-thymidine\;assay$), and protein and collagen assay(3H-proline assay) were carried out. In the MIT assay, after 200ul MTT solutionlconeentration of 5mg/ml) were added to the each well of the 24-well plates and incubated for 3 hours, and 200 ul DMSO were added so as to dissolve insoluble blue formazan crystals which was formed in incubated period. Then it read plates on a ELISA reader. For mitogenic assay, 1 uCi/ml $^3H-thymidine$ was added to each well for the final 2 hours of the incubation periods. After labeling, the wells were washed 3 times with ice cold PBS and 4 times with 5% TCA to remove unincorporated label and precipitate the cellular DNA. DNA, with the incorporated $^3H-thymidine$, was solubilized with 500 ul of 0.1% NaOH/0.1% SDS. A 250 ul aliquot was removed from each well and placed in a scintillation vial with 4ml of scintillation cocktail. Using an liguid scintillation counter, counts per minute(CPM) were determined for each samples. 3 uCi/ml $^3H-proline$ was added to each well for the final 4 hours of the incubation periods and total protein and percent collagen synthesis were carried out. The results indicate that minocycline treated group with $100{\mu}g/ml$ concentration for 1.5 hours significantly increased than that of control in cell attachment, and cell process is also evident compared with that of control in cell morphology, and the cellular activity and DNA synthesis rate of cells treated minocycline and $TGF-{\beta}1$ significantly increased than that of control values, but were below to values of the $TGF-{\beta}1$ only treated group in MIT assay and $^3H-thymidine\;assay$, and the total protein synthesis of minocycline and $TGF-{\beta}1$ treated group also significantly increased than that of control values, but the percent collagen synthesis of tested group significantly decreased to compared with control. On the above the findings, the tested group of minocycline and $TGF-{\beta}1$ did not increase the eff

성인성 치주염환자에서 분리한 Porphyromonas gingivalis와 Prevotella intennedia의 항생제 내성에 관한 연구

김흥식,유형근,신형식,Kim, Heung-Shik,You, Hyung-Keun,Shin, Hyung-Shik 대한치주과학회 1996 Journal of Periodontal & Implant Science Vol.26 No.3

The purpose of this study was to an in estimate the antibiotic susceptibility of P. gingivalis and P. interrnedia isolate from the subgingival plaque to adult periodontitis. Six P. gingivalis and five P. intermedia bacterial strains were tested for their susceptibility to 10 antimicrobial agents under disc diffusion method and broth dilution method. Ten patients with deep pocket(6mm) were selected for this study. They had not taken antibiotics for 6 months and no history of dental treatment for 6 months before this study. The result were as follow : 1. For antibiotic disc diffusion method, six P. gingivalis and five P. interrnedia were tested with 10 antimicrobial agents which comprised penicillin, gentamycin, clindamycin, lincomycin, ampicillin, erythromycin, tetracycline, amikacin, chloramphenicol, vancomycin. The sensitive antibiotics were tetracycline, penicillin, ampicillin, vancomycin, chloramphenicol and resisitent antibiotics were lincomycin. The other antimicrobial agents were less active. 2. From the study of determination on the minimal inhibitory concentration(MIC) by broth dilution method, the MIC of tetracycline to P. gingivalis and P. intermedia were $0.5-1.0{\mu}g/ml$, $0.5{\mu}g/ml$, that of ampicillin were $1-8{\mu}g/ml$, that of clindamycin were $1-32{\mu]g/ml$, $8-16{\mu}g/ml$, that of lincomycin were $16-32{\mu}g/ml$, $2-32{\mu}g/ml$. These data suggest that tetracycline and ampicillin may be valuable drug in the elemination of P. gingivalis and P. interrnedia from the patients with adult periodontitis.

Prostaglandin과 Dibutyryl cAMP가 조골세포의 활성과 파골세포 형성에 미치는 영향

목성규,유형근,신형식,Mok, Sung-Kyu,You, Hyung-Keun,Shin, Hyung-Shik 대한치주과학회 1996 Journal of Periodontal & Implant Science Vol.26 No.2

To maintain its functional integrity, bone is continuously remodelled by a process involving resorption by osteoeclasts and formation by osteoblasts, In order to respond to changes in the physical environment or to trauma with the relevant action, this process is strictly regulated by locally synthesized or systemic fators, Prostaglandin $E_2(PGE_2$) is perhaps one of the best studied factors, having been known to affect bone cell function for several decades.$PGE_2$ has both anabolic and catabolic activities. Excess of $PGE_2$ has been implicated in a number of pathological states associated with bone loss in a number of chronic inflammatory conditions such as periodontal disease and rheumatoid arthritis. $PGE_2$ and other arachidonic acid metabolites have been shown to be potent stimulators of osteoclastic bone resorption in organ culture. The anabolic effects of $PGE_2$ were first noticed when an increase in periosteal woven bone formation was seen after the infusion of $PGE_2$ into infants in order to prevent closure of the ductus arteriosus. The cellular basis for the catabolic actions of $PGE_2$ has been well characterized. $PGE_2$increases osteoclast recruitment in bone marrow cell cultures. Also $PGE_2$ has a direct action on osteoclast serving to inhibit activity and can also indirectly activate osteoclast via other cells in the vicinity, presumably osteoblast. The cellular mechanisms for the anabolic actions of $PGE_2$ are not nearly so well understood. The purpose of this paper was to study the effects of $PGE_2$ and dibutyl(DB)cAMP on osteoblastic clone MC3T3El cells and on the generation of osteoclasts from their precursor cells. The effect of $PGE_2$ and DBcAMP on the induction of alkaline phoaphatase(AlP) was investigated in osteoblastic clone MC3T3El cells cultured in medium containing 0.4% fetal bovine serum. $PGE_2$ and DBcAMP stimulated ALP activity and MTT assay in the cells in a dose-dependent manner at concentrations of lO-SOOng/ml. Cycloheximide, protein synthesis inhibitor, inhibited the stimulative effect of $PGE_2$ and DBcAMP on ALP activity in the cells. $PGE_2$also increased the intracellular cAMP content in a dose-dependent fashion with a maximal effect at 500ng/ml. The effect of $PGE_2$ on the generation of osteoclasts was investigated in a coculture system of mouse bone marrow cells with primary osteoblastic cells cultured in media containing 10% fetal bovine serum.After cultures, staining for tartrate-resistant acid phosphatase(TRAP)-marker enzyme of osteoclast was performed. The TRAP(+) multinucleated cells(MNCs), which have 3 or more nuclei, were counted. More TRAP(+) MNCs were formed in coculture system than in control group. $PGE_2(10^{-5}10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in culture. $PGE_2(10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in coculture of osteoblastic clone MC3T3E1 cells This results suggest that $PGE_2$ stimulates the differentiation of osteoblasts and generation of osteoclast, and are involved in bone formation, as well as in bone resorption.

성인형 치주염 환자에서 분린한 Actinobacillus actinomycetemcomitans의 항생재 내성에 관한 연구

나용철,유형근,신형식,Na, Yong-Cheol,You, Hyung-Keun,Sihn, Hyung-Sik 대한치주과학회 1996 Journal of Periodontal & Implant Science Vol.26 No.1

The purpose of this study was to an in estigate the antibiotic susceptibiliity of Actinoobacillus actinomycetemcomitans isolated from the subgingival plaque to adult periodontitis. Seven bacterial strains were tested for their susceptibility to 10 antimicrobial agents under disk diffusion method and broth dilution methold, Seven patients with deep pocket(6mm) were selected for this study. They had not taken antibiotics for 6 months and no history of dental treatment for 6 months and no history of dental treatment for 6 months before this study. The results were as follows : 1. For the antibiotic disk diffusion method, seven A. actinimycetemcomitans were tested with 10 antimicrobial agents which comprised penicillin, gentamycin, clindamycin, lincomycin, ampicillin, erythromycin, tetracycline, amikacin, chloramphenicol, and vancomycin. The sensitive antibiotics were tetracycline, vancomycin, and chloramphenicol and the resistant antibiotics were clindamycin, and lincomycin. The other antimicrobial agents were less active. 2. From the study of determination on the minimal inhibitory concentration(MIC) by broth dilution method, the MIC of tetracycline to seven strains of the A. actinomycetemcomitans was $0.5-1.0{\mu}g/ml$, that of clindamycin was $32{\mu}g/ml$. These data suggest that tetracycline may be valuable drugs in the elimination of A. actinimycetemcomitans from the patients with adult periodontitis

방사선 사진과 디지털 영상에서 실험적 치근 이개부 병소의 감지도에 관한 연구

이건일,유형근,신형식,Lee Geon-Il,You Hyung-Keun,Shin Hyung-Shik 대한영상치의학회 1995 Imaging Science in Dentistry Vol.25 No.1

The aim of this study was to evaluate clinician's detectability in the diagnosis of bone loss in the bifurcation of mandibular molars on periapical radiographs and Digital images. Periapical radiographs were obtained of the first molars in 2 dry mandibles after preparation of bony defects corresponding to degree I, degree II and degree III buccal furcation involvements. The radiographs were randomly presented to 39 clinicians(1 oral radiologist, 4 periodontist, 34 general dentists) who were asked to determine the presence or absence of bone loss. Periapical films were digitized with a TV camera. Digital images were assessed by 15 clinicians(1 oral radiologist, 4 periodontist, 10 general dentists). I. the overall diagnostic accuracy of Digital images for detection of bone loss in the bifurcation of mandibular molars was higher than that of the periapical radiographs. 2. the largest increase in diagnostic accuracy was found between lesion grade II and III on both radiographs and Digital images(P<0.05). 3. there was no significant difference between the standard state and the controlled contrast state on Digital images. 4. the overall diagnostic accuracy of I radiologist and 4 periodontists was better than that of the general dentists for detecting bifurcation involvements.