PDGF와 IGF-I 병용 사용시 치주인대세포의 증식과 세포활성에 미치는 영향에 관한 연구

서조영,신홍인,경희문,Suh, Jo-Young,Shin, Hong-In,Kyung, Hee-Moon 대한치주과학회 1996 Journal of Periodontal & Implant Science Vol.26 No.2

Current acceptable methods for promoting periodontal regeneration are based on removal of diseased soft tissue. root treatment, guided tissue regeneration, introduction of new graft materials and biological mediators. Insulin-like growth factor-I(IGF-I) and Platelet-derived growth factor-BB(PDGF-BB), the members of the polypeptuyde growth factor family have been reported as the biological mediators which regulate a variety cellular matrix biologic activities of wound healing process including the cell proliferation, migration and extracellular matrix synthesis.The purposes of this study is to evaluate the combination effects of IGF-I and PDGF-BB on the cellular activity of the periodontal ligament cells to act as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM containing 10% FBS at the $37^{\circ}C$, 5% CO2 incubator. Author measured the DNA synthetic activity, and total protein, collagen and noncollagenous protein synthetic activities according to the concentration of 10,100ng/ml IGF-I and1,10 ng/ml PDGF-BB in combination. The results were as follows: Significantly increased in the 1 ng/ml PDGF-BB alone compared to the 10 ng/ml PDGF-BB alone(P<0.01) and in the 1 ng/ml PDGF-BB and 10, 100ng/ml IGF-I in combination compared to the 1 ng/ml PDGF-BB alone(P<0.05, P<0.0l). The synthetic activity of the total protein and collagen is significantly increased like to the synthetic activity of the DNA(P<0.05). The synthetic activity of the noncollagenous protein is increased according to the concentration of IGF_I, but not statistically statistically significant(P>0.05). The percent of the collagen is significantly in the 1ng/ml PDGF-BB and 10ng/ml IGF-I in combination compared to the 1ng/ml PDGF-BB alone(P<0.05) and in the 10ng/ml IGF-I in combination compared to the 10ng/ml PDGF-BB alone(P<0.05). The synthetic activity of the DNA is In conclusions, the percent study shows that PDGF-BB and IGF-I in combination have a potentiality to enhance the DNA synthesis and the total protein and collagen synthesis of The periodontal ligament cells, especially it is more significant in the low concentration of PDGF-BB compared to the high one. Thus, the PDGF-BB and IGF-I in combination may have important roles in promotion of periodontal litgment healing, and consequently, may useful for clinical application in periodontal regenerative procedures.

치주인대세포의 부착과 전개에 관한 형태학적 관찰

이진미,서조영,박준봉,Lee, Jin-Mi,Suh, Jo-Young,Park, Joon-Bong 대한치주과학회 1993 Journal of Periodontal & Implant Science Vol.23 No.1

치주조직의 초기치유과정에서 가장 중요한 역할을 하는 치주인대세포의 부착과 전개에 대한 형태학적 관찰을 위하여 교정치료 목적으로 발거된 치아로부터 치주인대조직을 채취하여 초기배양하고 동일한 양($5{\times}10^3ml$)의 치주인대 세포를, 2장의 유리가 포함된 35mm 배양접시에 접종한 후, $CO_2$ 배양기에서 배양하여 세포배양개시후 10분, 30분, 90분, 6시간, 12시간, 24시간의 시간간격에 따라서 광학위상차현미경과 주사전자현미경을 이용하여 세포부착과 전개양상을 관찰한 결과 다음과 같은 결과률 얻었다. 10분간 배양한 세포는 구형을 나타내었으며, 기질측의 세포질이 박판엽상으로 확장되어 기질에 부착개시한 양상이 보였다. 배양 30분 후 판모양의 세포돌기인 박판상돌기에서 세사상돌기롤 내어 부착하는 양상이 관찰되었고, 세포표면은 소기포와 미세융모로 덮여 있었다. 90분간 배양한 세포에서 핵은 세포의 중심부에 위치하였고 세포질이 방사선상으로 확장되어 부착된 양상을 보였으며, 미전개된 세포중심부에는 미세융모와 소기포로 덮여져 있었다. 배양 6시간 후 세포는 신장된 양상을 보였고 타원형의 핵이 관찰되었다. 세포의 양극에서 박판상돌기가 관찰되었고 다시 세사상돌기가 분지되는 양상을 보였는데 이 돌기의 말단부는 팽대되어 기질에 부착하였다. 12시간 배양한 후의 치주인대 세포는 뚜렷한극성화를 보이면서 길게 신장된 양상을 나타내었다. 배양 24시간 경과후 신장된 양상의 핵이 세포의 한쪽극에 치우쳐 있었으며, 세포외형은 길게 신장된 방추상의 형태가 관찰되었다. One of the important initial events required for periodontal regeneration is the attachment and subsequent spreading of periodontal ligament cells on the root surface. The purposes of this study is to investigate the attachment and spreading pattern of human periodontal ligament cell on the surface of glass slides. After establishment of a cell line of the primary cell culture from the periodontal ligament of 1st premolar teeth which were extracted for the purpose of orthodontic treatment, author dispersed the cells at $5{\times}10^3\;cells/ml$ into the each 35mm culture petri-dish containing 2 glass slides. To observe the morphological changes of the cells which attached to the surfaces of glasses at every designed time schedule, author used the inverted phase contrast microscope and scanning electron microscope. During the whole experiment culture condition was at $37^{\circ}C$, 100% Humidity, 5% $CO_2$ gas incubator. The following results were obtained. Periodontal ligament cells showed spherical outline and started to attach to glass surface by basal sytoplasmic extension after 10min in culture. After 30min in culture, periodontal ligament cells were attached to glass surface by well - developed filopodia which protruded from the lamellipodia. The cell surface is covered with bubble-like structures and occasional microvillus can be seen with diffculty among these structures. After 1.5hr in culture, peridontal ligament cells shhowed radially well-spread cytoplasm and the nucleus was centered on its cytoplasm. Unspread central region of the cell was covered with numerous microvilli. The change of cell attachment and spreading pattern was manifest at 6hr in culture. At this time, periodontal ligament cell showed elongated outline and an oval-shaped nucleus. After 12hr in culture, periodontal ligament cells showed more stretched fibroblast-like appearance with polarity. Two long lamellipodia can be seen around the both terminal ends of cells. After 24hr in culture, periodontal ligament cells showed spindle shapes and an oval-shaped nucleus was slanted toward one side of the cell.

치근면 구연산 도포가 치주인대세포의 부착과 전개에 미치는 영향

이상구,서조영,박준봉,Lee, Sang-Gu,Suh, Jo-Young,Park, Joon-Bong 대한치주과학회 1993 Journal of Periodontal & Implant Science Vol.23 No.1

정상치아와 이환치아 및 구연산에 의하여 탈회한 치근면에대한 치주인대세포의 부착상태를 비교관찰 하기 위하여 정상치아와 이환치아를 취하여 정상군, 이환치근의 치근면활택술군 및 구연산처리군으로 분류하고 시험관적 실험을 통하여 관찰하였다. 세포부착실험전 각 군의 치근면의 형태를 주사전자현미경을 이용하여 관찰하였으며, 사람의 정상소구치에서부터 얻어진 치주인대세포를 배야하여 각 군의 절편을 함유한 조직배양기에 ml당 $4.5{\times}\;10^4$개의 세포를 가진 배양액 1ml씩을 넣고 동일조건하에서 30분, 1시간, 2시간, 6시간, 12시간 및 24시간 동안 배양한 후 세포의 부착이 일어난 후 상태를 주사전자현미경을 이용하여 관찰하였고, 세포증식정도를 알아보기 위하여 각 절편에 $5{\times}10^4$개의 세포가 함유된 배양액 1ml씩을 넣고 6시간동안 배양 후, 새 배양기에 옮기고 24, 48, 72시간동안 배양하여 trypsin 처리로 세포를 분리시킨 후 광학위상차 현미경을 이용, 치근단위면적당 증식된 세포수를 측정하여 다음과 같은 결과를 얻었다. 세포부착실험전 각 군의 치근면 형태를 관찰하였을 때 구연산처리군에서는 교원섬유의 노출부위라고 여겨지는 미세한 돌출구조들을 관찰할 수 있었으며 상아세관의 노출부위라고 여겨지는 함몰양상이 치근활택술군에 비하여 현저하였고 정상치아군에서는 이러한 양상을 관찰할 수 없었다. 실험 각 군 및 대조군 사이에서 세포들의 형태적인 차이점은 분명하지 않았으나 구연산처리군에서 다른 군들보다 초기의 세포부착양상이 다소 빠르게 진행 됨이 관찰되었다. 세포배양 개시 후 6시간이 경과 한 후에는 모든 군에서 공히 세포의 형태가 초기의 구형의 형태에서 편평한 세포의 형태로 변화되기 시작함이 관찰 되었고 24시간 이후에는 모든 군에서 세포들이 치근면에 납작하게 부착되고 일반적인 섬 유아세포의 형태로 변화되어 셰포의 전개가 완성된 양상이 보였다. 세포증식율의 실험에서는 24시간 후 증식된 세포 수는 치근면활택술군에서 가장높게 나타났으며 정상군에서 가장 낮은 수치를 보였으며 48시간 및 72시간후 측정에서는 구연산처리군이 다른 군들에 비해 더 많은 수의 세포증식을 관찰할 수 있었으며 각 군간의 차이는 통계학적으로 유의하였다. This in vitro study was undertaken to observe whether citric acid application aids the attachment and proliferation of human periodontal ligament cells to the root surfaces of periodontally diseased teeth. The roots were prepared so that the comparison could be made among the control healthy root surface, citric acid demineralized and non-demineralized root planted surfaces. Prior to the cell attachment experiment, each groups were prepared for scanning electron microscopic (SEM) examinations of root surface morphology, All specimens were fixed with phosphate buffered glutaraldehydes, postfixed with phosphate buffered osmium tetraoxide and stained with phosphate buffered tannic acid. dehydrated in ethanol, critical point dried, sputter coated with gold and examined under the SEM. In the cell attachement experiment, human cultured periodontal ligament cells at concentration to $4.5{\times}\;10^4\;cells/ml$ were seeded in each culture well which contained prepared roots and incubated for 30min 1, 2, 6, 12 and 24 hours at 37, 5% $CO_2$air incubator. Than the specimens were prepared for SEM examination using, the same methods as described above. In the cell proliferation experiment, $5{\times}\;10^4\;cells/ml$ cells were seeded incubated with the specimens for 6 hours. Then, all of the specimens were moved into fresh culture well and incubated for 24, 48, and 72 hours. The cell counting was done after trypsinization, under light microscope. The results were as follows. When viewed the surface morphology prior to the cell attachment, the non acid treated root planed surface displayed scaling striation and occasional bacteria and calculus. The citric acid treated specimens displayed little debris on the surface and funnel shaped orifices of dentinal tubules. There were no apparent differences in the morphology of cells attached to the control and experiment groups. However, in initial attachement, there was a slight more enhanced appearance in attachment in citric acid treated groups than other root surfaces. After 6 hours of incubation, most of the cells initiated the alteration of cell morphology from ovoid to spindle shapes. After 24 hours of incubation, most of the cells displayed proliferated appearance and connected with each other via numerous processes. In the cell proliferation experiments, there were statistically significant increased number of cells in citic acid treated groups than other groups.

BMP-교원질 섬유막 복합체에 의한 이소성 골형성

신홍인,서조영,Shin, Hong-In,Suh, Jo-Young 대한치주과학회 1996 Journal of Periodontal & Implant Science Vol.26 No.1

To investigate the efficiency of a fibrous collagen membrane(FCM) composed of bovine skin type I atelocollagen as a carrier for BMP, partially purified bovine BMP/FCM($0.3mg/10{\times}5{\times}1mm$) composites were implanted into the dorsal subcutaneous tissue of rats. FCM alone was also implanted as a control. The implants were harvested at 1, 2, 3, and 10 weeks after implantation, then prepared for routine light microscopic observation. The FCM alone did not induce osteogenesis and revealed no specific foreign body reaction nor was there any definite resorptive evidence for 10 weeks after implantation, while the BMP/FCM composites induced favorable bone formation in a process that resembled an endochondral and direct ossification mode. At 10 weeks, the well formed bone confined to embedded collagen fibers revealed hematopoietic marrow between bony trabeculae without evidence of resorptive or degenerative changes . These findings support the suggestion that BMP may induce undifferentiated mesenchymal cells into either chondroblasts or osteoblasts or both independantly according to the chemico- physical characteristics of the carrier, which develops the endochondral and/or direct bone formation process, and suggest that the FCM may be a favorable carrier for BMP.

Histological Observations on Bone Healing with Bioactive Glass in Horizontal Ridge Augmentation;A Report of Four Cases

박진우,서조영,Park, Jin-Woo,Suh, Jo-Young The Korean Academy of Periodontoloy 2006 Journal of Periodontal & Implant Science Vol.36 No.3

임프란트 식립을 필요로 하는 환자의 수평적 치조제 결손의 증대를 위해 골유도재생술과 병용한 bioactive glass (BG) $(Biogran^{(R)})$ 이식의 골재생 양상을 각기 다른 치유기간을 부여한 4명의 환자에서 평가하였다. 6, 8, 10, 18개월의 치유기간 후 임프란트 식립부위에서 조직절편을 채득하여 골재생을 조직계측학적으로 평가하였다. 임프란트 식립을 위한 surgical reentry시 모든 이식부위는 임상적으로 명확한 수평적 치조제 폭경의 증가를 관찰할 수 있었다. 하지만 조직학적 분석결과 BG는 불량한 골전도성을 나타내었다. 6, 8개월의 치유기간후, 이식부위에서 신생골이 거의 관찰되지 않았으며(2.5%이하), 이식부와 기존 골의 경계부위에서 BG particle에 대한 신생골 성장과 결합양상 또는 관찰할 수 없었다. 10개월의 치유기간후 기존 골조직으로부터 성장한 신생골의 BG particle과의 직접적인 접촉양상을 일부 관찰할 수 있었다. 이식부는 13.2%의 광물화된 신생골조직을 보였고, 대부분의 BG particle은 결체조직으로 둘러싸여 있었다. 18개월의 치유기간이 부여된 환자의 조직절편에서 신생골은 이식부의 10.7%를 차지하여 비교적 낮은 신생골 형성양을 나타내었고, 이식부에 존재하는 잔존BG particle은 대부분은 결체조직으로, 일부분에서 광물화된 골조직으로 둘러싸여 있었다. 6, 8, 10, 18개월에서 잔존 BG particle양은 전체 이식부 면적에 대해서 각기 22.3%, 26.5%, 30.7%, 18.7%로 나타났다. 본 증례보고는 비록 한정적인 4명의 환자에서의 조직계측학적 평가결과이지만, 수평적 치조제 결손의 증대를 위해 골유도재생술과 병용한 bioactive glass이식은 불량한 골전도성으로 인해 효과적인 골재생을 위한 이식재로서는 적절하지 않을 수 있음을 나타낸다.

만성 치주염 환자에서 2형 당뇨병이 $PGE_2$, MMP-14 및 TIMP의 발현양상에 미치는 영향

김선우,박진우,서조영,이재목,Kim, Seon-Woo,Park, Jin-Woo,Suh, Jo-Young,Lee, Jae-Mok 대한치주과학회 2007 Journal of Periodontal & Implant Science Vol.37 No.4

Purpose: The purposes of this study were to compare and quantify the expression of $PGE_2$, MMP-14 and TIMP-1 in the gingival tissues of patients with type 2 diabetes mellitus and healthy adults of chronic periodontitis with alveolar bone resorption. Material and methods: Gingival tissue samples were obtained during periodontal surgery or tooth extraction. According to the patient's systemic condition & clinical criteria of gingiva, each gingival sample was devided into three groups. Group 1 (n=8) is clinically healthy gingiva without bleeding and no evidence of bone resorption or periodontal pockets, obtained from systemically healthy 8 patients. Group 2 (n=8) is inflammed gingiva from patients of chronic periodontitis with alveolar bone resorption. Group 3(n=8) is inflammed gingiva from patients of chronic periodontitis with alveolar bone resorption associated with type 2 diabetes. Tissue samples were prepared and analyzed by Western blotting. The quantification of $PGE_2$ MMP-14 and TIMP-1 were performed using a densitometer and statistically analyzed by one-way ANOVA followed by Tukey test. Results: The expressions of MMP-14 and TIMP-1 were showed increasing tendency in group 2 & 3 compared to group 1. The expressions of $PGE_2$, MMP-14 were showed increasing tendency in group 3 compared to group 1 and group 2. According to MMP-14 levels were increasing, $PGE_2$ showed increasing tendency in group 3, and although $PGE_2$, MMP-14 levels were increasing, TIMP-1 levels were similar expressed comparing to group 2. Conclusion: In conclusion, this study demonstrated that the expression levels of MMP-14 and TIMP-1 had increasing tendency in inflammed tissue. It can be assumed that $PGE_2$ and MMP-14 may be partly involved in alveolar bone resorptive process and the progression of periodontal inflammation associated to type 2 DM.

이개부 병변을 가진 하악 대구치의 치료에 있어 치근절제술과 터널화의 임상결과 비교

김철우,박진우,서조영,이재목,Kim, Cheol-Woo,Park, Jin-Woo,Suh, Jo-Young,Lee, Jae-Mok 대한치주과학회 2009 Journal of Periodontal & Implant Science Vol.39 No.1

Purpose: The resective treatment in mandibular Degree III furcation areas includes root resection and tunnel preparation. The purpose of this retrospective study is to evaluate which treatment procedure(root resection vs tunnel preparation) has more favorable prognosis in mandibular Degree III furcation involvement. Materials and methods: The subjects of this study were patients who were treated their mandibular posteriors with either root resection(22 teeth on 22 patients) or tunnel preparation(24 teeth on 22 patients) and have participated in periodic recall check program for at least 2 years. Failure rate, plaque index, bleeding on probing and probing pocket depth were measured and the results were compared and statistically analysed. Results: Failure rates of root resection and tunnel praparation were 13.6% and 16.7%, respectively, and showed no significant difference between two procedures. No significant differences were observed between two procedures with respect to plaque index, bleeding on probing and probing pocket depth. Conclusion: root resection and tunnel preparation are both effective procedures to treat mandibular posterior furcation involvement, if adequate treatment of choice is made and patient's cooperation is accompanied. However, tunnel preparation is more preservative procedure in mandibular posterior furcation involvement since root canal treatment and prosthetic restoration are needed additionally, in case of root resection.

치근이개부 II급 병변에서 조직유도재생술의 임상적 치유양상의 비교

문선영,이재목,서조영,Moon, Sun-Young,Lee, Jae-Mok,Suh, Jo-Young 대한치주과학회 1999 Journal of Periodontal & Implant Science Vol.29 No.3

The purpose of this study is to compare the healing aspects of the use of ePTFE membrane alone versus combination treatment of ePTFE membrane and bone grafts on class II furcation defects. Seventeen defects were applied ePTFE membrane alone on mxillary molar buccal class II furcation defects as Group I, seventeen defects were applied ePTFE membrane and bone grafts on maxillary molar buccal class II furcation defects as Group II, twenty-three defects were applied ePTFE membrane alone on mandibular molar buccal class II furcation defects as Group III, twenty defects were applied ePTFE membrane and bone grafts on mandibular molar buccal class II furcation defects as Group IV . Measurements were made to determine clinical attachment level, probing depth, gingival depth, SBI, mobility at baseline, 3, 6, 12 months postoperatively. Additional measurements were made to determine membrane exposure level at surgery, 1, 2, 6 weeks postoperatively. And then healing patterns and postoperative complications were evaluated. The result as follows : There were statistically significant differences in probing depth reduction, clinical attachment gain, mobility reduction at values of 3, 6, 12 months postoperatively compared to values of baseline(p<0.05), whereas no significant differences in SBI and gingival recession. In group II, membrane exposure level was increased at 1, 2, 6 weeks postoperatively compared to value of baseline(p<0.05). There were statistically significant differences in changes of probing depth at 3, 6, 12 months postoperatively in combination groups of ePTFE membrane and bone graft compared to groups of ePTFE membrane alone(p<0.05). The vast majority of cases fall into typical healing and delayed healing response when membranes were removed in all groups. Pain and swelling were common postoperative complications. In conclusion, this study was showed more effective healing aspects in combination treatment of ePTFE membrane and bone graft than ePTFE membrane alone and on mandibular molar class II furcation defects than maxillary molar.

골연하 결손부에서 조직유도재생술의 장기적 방사선학적 변화 관찰

최미혜,박진우,서조영,이재목,Choi, Mi-Hye,Park, Jin-Woo,Suh, Jo-Young,Lee, Jae-Mok 대한치주과학회 2007 Journal of Periodontal & Implant Science Vol.37 No.2

Periodontal surgery as part of the treatment of periodontal disease is mainly performed 1) to gain access to diseased areas for adequate cleaning; 2) to achieve pocket reduction or elimination; and 3) to restore the periodontal tissues lost through the disease; i.e., a new attachment formation of periodontal regeneration. To accomplish the latter, often referred to as the ultimate goal of periodontal therapy, a number of surgical procedures have been advocated throughout the years. Clinical studies have demonstrated that considerable gain of clinical attachment and bone can be achieved following guided tissue regeneration (GTR) therapy of intrabony defects. The aim of this study was to analyse the radiographic bone changes 2-year after GTR using a bone graft material and nonresorbable membrane. Patients attending the department of periodontics of Kyungpook National University Hospital were studied. Patients had clinical and radiographic evidence of intrabony defect(s), 33 sites of 30 patients aged 32 to 56 (mean age 45.6) were treated by GTR with a bone graft material and nonresorbable membrane. Baseline and 2-year follow-up radiographs were collected and evaluated for this study. Radiographic assessment includes a bone fill, bone crest change, defect resolution, and % of defect resolution. Pre- and post-treatment differences between variables (maxilla and mandible, defect depth, defect angle, bone graft materials) using the paired t-test were examined. We observed $2.86{\pm}1,87mm$ of bone fill, $065{\pm}0.79mm$ of crestal resorption, $3.49{\pm}2.11mm$ of defect resolution, and $44.42{\pm}19.51%$ of percentage of defect resolution. Mandible, deeper initial defect depth, narrower initial defect angle showed greater bone fill, defect resolution, and % of defect resolution. But no difference was observed between xenograft and allograft. Outcome of GTR as a therapy of intrabony defect was better than other therapy, but herein, good oral hygiene maintenance as a anti-infective treatment and periodic recall check of patients are essential.

Establishment of B-1 cell-derived polyreactive monoclonal antibodies and expression of costimulators by B-cell to antigenic stimulation

이주연,최점일,서조영,Lee, Ju-Youn,Choi, Jeom-Il,Suh, Jo-Young The Korean Academy of Periodontoloy 2007 Journal of Periodontal & Implant Science Vol.37 No.2

연구 목적 : 면역 반응에서 B-1 세포의 정확한 역할은 아직 명확히 규명되지 않았으나 최근 B-1 세포가 면역의 내성을 야기하고 유지하는데 필요한 특성들을 가지고 있음이 밝혀지고 있다. 이에 본 연구에서는 B-1 세포 유래 단클론 항체의 특성과 항원 자극에 대한 B 세포의 동시자극자 (MHC Class, B7-2, 7-2) 발현을 평가하여 B 세포의 면역조절 기능을 알아보고자 한다. 연구 대상 및 방법 : B-1 세포 유래 단클론 항체를 형성하는 잡종세포주를 이용하여 다양한 내, 외인성 항원에 대한 단클론 항체의 반응 양상을 평가하였다. 여러 내, 외인성 항원으로 면역한 쥐의 복강과 비장 B 세포의 동시자극자의 발현을 Fluorescence Activated Cell Sorting (FACS)을 이용하여 평가하였다. 결과 : 최종적으로 단클론을 형성하는 2개의 클론을 형성하였고, 이 B-1 세포 유래 단클론 항체는 dose-saturable pattern을 띄는 다중 반응성을 나타내었다. FACS를 이용한 동시자극자의 발현 검사에서는 MHC 발현은 복강과 비장의 B 세포가 유사하였으나, B7-1과 7-2는 복강의 B 세포에서 더 뚜렷한 발현을 보여주었다. 결론 : B-1 세포 유래 단클론 항체는 다양한 내, 외인성 항원 자극에 대해 dose-saturable한 다중반응성을 나타낸다. 복강과 비장의 B세포는 내, 외인성 항원의 면역에 있어서 동시자극자 발현이 명확히 다른 양상을 나타냄을 확인할 수 있었다.