• Journal of Periodontal and Implant Science
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• v.26 no.4
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• pp.907-931
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• 1996

The purpose of this study was to study of the effects of the bioglass and the natural coral on healing process of the alveolar bone defects. Three adult dogs aged 1 to 2 years were used in this study. Experimental alveolar bone defects were created surgically with surgical bur and bone chisel at the furcation area of the buccal surface of the right and left mandibular 3rd, 4th premolars. Twelve experimental alveolar bone defects were devided into four groups according to the type of graft materials. The groups were as follows : 1. flap operation with root planing & curettage(Negative control group) 2. flap operation with autogenous bone(Positive control group) 3. flap operation with bioglass(BG group) 4. flap operation with natural coral(NC group) At 2, 4, and 8 weeks, the dogs were serially sacrificed and specimens were prepared with Hematoxylin-Eosin stain for light microscopic evaluation. The results of this study were as follows : 1. The defect areas were filled with granulation tissue at two weeks in negative control group. But in other groups, the appearance of connective tissues around graft materials were formed more densely and the response of inflammation by graft materials itself was not found. 2. In every control and experimental groups at two weeks, there was seen the accumulation of the formation of new bone trabeculae at the bottom of defects and gradually expanded toward the graft materials and in autogenous group there was slightly seen the formation of new cementum. 3. There was seen the erosion of central portion of bioglass particles at two weeks in BG group, and the erosion of the central portion was developed more progressively and was filled with bone-like tissues at eight weeks. 4. The natural coral particles were encapsulated by densely connective tissues and seen the formation of new bone tissues at four weeks and developed more new bone and cementum formation at eight weeks. From the results of this study, the bioglass and the natural coral may be biocompatible and have a weak adverse reaction to the periodontal tissues.

• Journal of Periodontal and Implant Science
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• v.25 no.3
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• pp.539-556
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• 1995

Periodontal therapeutic modalities should be re-establishing and regenerating the periodontal tissue previously lost to the disease. To achieve periodontal regeneration, periodontal ligament cells must selective migrate to the deneded root surface, attached and proliferated it. Local pH concentration is one of the most factors that periodontal regeneration. The aims of this study were to examine on biological effects of pH to the human periodontal ligament cells in vitro, especially on the cell morphology, attachment, activity, vitality and viability. Human periodontal ligament cells were cultured from extracted tooth for non-periodontal reason. Immediately after extraction, any soft tissue adhering to the cervical parts of the roots was carefully removed with a sterile curette. To produce different pH levels in the media, Eagle's MEM was adjusted from pH 6.6 to 8.2 in 0.2 intervals with 1 M NaOH and 1 N HCl. After cultivation, Then, Periodontal ligament cells were cultured at pH ranging from 6.6-8.2. attachment assay was done at 1, 2 day incubation and activity assay was done at 1, 2, 3 day incubation. The experiments were evaluated by scaning electron microscopic techniques (HITACHIX-650 Scaning Electron Microanalyzer, Tokyo, Japan), MTT assay, and the cultured periodontal ligament cells were fixed in neutral formalin for 24 hours and immunohistochemically processed by PCNA for proliferating ability. The surviving cells in the medium showed slightly increased volume and widening intercellular distances at low concentration of pH than control group (pH 7.4), and apparently shrinkage at high concentration of pH than control group (pH 7.4). The results of the statistical analysis from the experiment on attachment, vitality and viability were as follows. Attachment of periodontal ligament cells at 1st and 2nd day, similar attachment rate of low concentration pH compared with control value (pH 7.4). But above pH 8.0, attachment rate were statistically significant decrease from control value(P<0.05). Periodontal ligament cell's activities were maximum at pH 7.6 by MTT assay. Similar with control value at low concentration of pH. But, the activities were statistically significant decrease at high concentraration of pH(P<0.05). Cellular proliferating rate (PCNA index) were statistically significant decrease from control value at low and high concentration of pH(p<0.05). This results suggested that hjgh concentration pH, in other words, alkali pH was cytotoxic effects on human periodontal ligament cells in vitro.

• Journal of Periodontal and Implant Science
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• v.25 no.3
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• pp.703-719
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• 1995

The purpose of this study was to observe the effects of the periodontal ligament on the healing and the formation of alveolar bone in the extraction socket, when this ligament had artificially remained in the socket during the tooth removal. Twenty rats aged 4 weeks were used and devided into the control groups (10) and the experimental groups (10) in this study. The maxillary right and left first molars were extracted in both groups. In the experimental groups the periodontal ligament was remained in the extraction sockets using 0.4% ${\beta}-aminopropionitrile$, and in the control the periodontal ligament was completely removed by curettage. At 1, 3, 5, 7 and 14 days after the tooth extraction, rats in both groups were serially sacrificed. And the specimens were prepared with Hematoxylin-Eosin stain for the light microscopic evaluation. The results of this study were as follows ; 1. On 1 day, the periodontal ligament was only found in the extraction socket walls of the experimental groups, and there was not the distinguishable difference between the control and the experimental groups. 2. On 3 days, there were more collagen fibers and the appearance of higher cellular density in the experimental groups than in the control. And the cells and collagen of the periodontal ligament were so actively proliferated and synthesized that invaded into the connective tissue of the extraction sockets in the experimental groups. 3. In the experimental groups, the trabecular bone was formed on the basal and lateral bone surface on 5 days. However, there was not the new bone forming appearance in the control groups at this time. 4. On 7 days, the trabecular bone was formed in the control groups. 5. On 14 days, the extraction sockets were almost entirely filled with the bony trabeculae in both groups. But, compared to the control group, the experimental groups showed the prominent differences in the amount & the density of the new bone formed. In conclusion, it was suggested that the residual periodontal ligament tissue in the extraction socket will play a major role as the important cell source in the healing and the new bone formation of the extraction socket.

• Journal of Periodontal and Implant Science
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• v.27 no.2
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• pp.317-328
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• 1997

Since laser therapy has been applied to dentistry, many dental practitioners are very interested in laser therapy on various intraoral soft tissue lesions including gingival hyperplasia and aphthous ulcer. The purpose of the present study was to determine the therapeutic effect and the harmful effect of a pulsed-Nd:YAG laser irradiation on human gingival tissue. In twenty periodontal patients with gingival enlargement, the facial gingival surface of maxillary anterior teeth was randomly irradiated at various power of 1.0W(100mJ, 10Hz), 3.0W(100mJ, 30Hz) and 6.0W(l50mJ, 40Hz) for 60 seconds by contact delivery of a pulsed-Nd:YAG laser(EN.EL.EN060, Italy). Immediately after laser irradiation, the gingival tissues were surgically excised and prepared in size of 1mm3. Subsequently the specimens were processed for prefixation and postfixation, embedded with epon mixture, sectioned in $1{\mu}$ thickness, stained with uranyl acetate and lead citrate, and observed under transmission electron microscope(JEM 100 CXII). Following findings were observed; l. In the gingival specimens irradiated with l.OW power, widening of intercelluar space and minute vesicle formation along the widened intercellular space were noted at the epithelial cells adjacent to irradiated area. 2. In the gingival specimens irradiated with 3.0W power, the disruption of cellular membrane, aggregation of cytoplasm, and loss of intercellular space were observed at the epithelial cells adjacent to irradiated area. 3. In the gingival specimens irradiated with 6.0W power, the disruption of nuclear and cellular membrane was observed at the epithelial cells adjacent to irradiated area. The ultrastructural findings of this study suggest that surgical application of a pulsed-Nd:YAG laser on human gingival tissue may lead somewhat delayed wound healing due to damage of epithelial cells adjacent to irradiated area.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.30 no.2
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• pp.142-149
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• 2008

The purpose of this study was to evaluate histomorphometrically a toothash - plaster of Paris mixture associated with collagen membrane ($Bio-Gide^{(R)}$), regarding new bone formation in the peri-implantitis defects in dogs. Three mandibular molars were removed from 1-year-old mongrel dogs. After 2 months of healing, 2 titanium implants with sandblasted with large grit and acid etched (SLA) surface were installed in each side of the mandible. Experimental peri-implantitis was induced with ligatures after successful osseointegration. Ligatures were removed after identification of bone defect beneath the level of 5th thread of fixture on radiographic image. The mucoperiosteal flaps were elevated and the contaminated fixtures were treated with chlorhexidine and saline. The bone defects were assigned to one of the following treatments: no guided bone regeneration (GBR) procedure (group 1), GBR with Bio-$Oss^{(R)}$ and Bio-$Gide^{(R)}$ (group 2), or GBR with toothash - plaster of Paris mixture (TPM) and Bio-$Gide^{(R)}$ (group 3). The dogs were sacrificed after 8 or 16 months. The mean percentages of new bone formation within the limits of the 5 most coronal threads were $17.83{\pm}10.69$ (8 weeks) and $20.13{\pm}13.65$ (16 weeks) in group 1, $34.25{\pm}13.32$ (8 weeks) and $36.33{\pm}14.21$ (16 weeks) in group 2, and $46.33{\pm}18.39$ (8 weeks) and $48.00{\pm}17.78$ (16 weeks) in group 3, respectively. The present study confirmed statistically considerable new bone formation within the threads in group 3 compared with group 1 at 8 and 16 weeks (P<0.05). Although, data analysis did not reveal significant differences between group 2 and 3, the latter showed better results during the period of 8 or 16 weeks. Our findings support the effectiveness of TPM as a GBR material in the treatment of peri-implantitis bone defect.

• Journal of Periodontal and Implant Science
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• v.29 no.4
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• pp.821-838
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• 1999

On the basis of the evidences that electrical stimulation could enhance proliferation and differentiation of bone cells and promote healing and regeneration of bone, this study was performed to investigate the effects of electrical stimulation on human periodontal ligament cells and gingival fibroblasts in vitro, which also have important roles in regeneration of periodontium, and to evaluate the potential of clinical application of electrical stimulation. Human periodontal ligament cells and gingival fibroblasts were primarily cultured from the root surface of extracted premolar and the adjacent gingiva without periodontal diseases. In control group, the cells ($5{\times}10^4$ cells/ml)were incubated only in Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum. In test groups, electrical stimulation was given at the current intensity of $0.25{\mu]A$(test group 1), $1.0{\mu}A$(test group 2), and $2.5{\mu}A$(test group 3) for 12 hours to the same culture media with the control group. After 12 hour exposure of electrical stimulation, the cells were incubated for 2 and a half days(60 hours), and then each group of cells was analyzed for cell proliferation, protein level, and activity of alkaline phosphatase. The results were as follows ; 1. The Rate of cell proliferation of every test group increased significantly in both periodontal ligament cells and gingival fibroblasts, and in periodontal ligament cells, test group 3 showed significantly increased proliferation compared to the other test groups(p<0.05). 2. In the protein levels, neither periodontal ligament cell nor gingival fibroblast showed statistically significant differences between control and test groups. 3. The activity of alkaline phosphatase in periodontal ligament cells increased significantly in all test groups(p<0.05), but there were no significant differences between 3 test groups. In gingival fibroblasts, the activity of alkaline phosphatase increased significantly only in test group 3(p<0.05). From the above results, it is concluded that electrical stimulation may have beneficial effects on the regeneration of destructed periodontal tissue in regard of the stimulation of periodontal ligament cells and gingival fibroblasts as well as electrically stimulated bone formation that has been known, and that electrical stimulation may have the potential of clinical application.

• Journal of Periodontal and Implant Science
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• v.32 no.3
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• pp.489-500
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• 2002

The purpose of this study was to evaluate newly fabricated tricalcium phosphate(TCP)/chitosan microgranuls as bone substitutes. TCP/chitosan microgranules were fabricated by dropping TCP-chitosan suspension into the NaOH/ethanol solution. The size of microgranules could be controllable via airflow rate. PDGF-BB was loaded into the fabricated granules via freeze-drying methods(300 ng/20 mg). To evaluate cell proliferation, cultured osteoblasts cell lines(MC3T3-El) was dropped on the BioOss(R), chitosan microgranules, TCP/chitosan microgranules and cultured for 1, 7 , 14, and 28 days. Scanning electron microscopic observation was done after 7 days of culture and light microscopic examination was done after 28 days of culture. PDGF-BB release from the microgranules was tested. Rabbit calvarial defects(8 mm in diameter) were formed and chitosan, TCP/chitosan, PDGF-TCP/chitosan microgranules, and BioGran(R) were grafted to test the ability of new bone formation. At SEM view, the size of prepared microgranules was 250-1000 um and TCP powders were observed at the surface of TCP/chitosan microgranules. TCP powders gave roughness to the granules and this might help the attachment of osteoblasts. The pores formed between microgranules might be able to allow new bone ingrowth and vascularization. There were no significant differences in cell number among BioOss(R) and two microgranules at 28 day. Light and scanning electron microscopic examination showed that seeded osteoblastic cells were well attached to TCP/chitosan microgranules and proliferated in a multi-layer. PDGF-BB released from TCP/chitosan microgranules was at therapeutic concentration for at least 1 week. In rabbit calvarial defect models, PDGF-TCP/chitosan microgranules grafted sites showed thicker bone trabeculae pattern and faster bone maturation than others. These results suggested that the TCP/chitosan microgranules showed the potential as bone substitutes.

• Journal of Periodontal and Implant Science
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• v.29 no.4
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• pp.803-823
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• 1999

In order to reveal immunopathogenesis of periodontal tissue destruction, it is important to clarify the molecular mechanism of trafficking and retention of activated leukocytes, including monocytes/macrophages. Gingival fibroblasts may be involved in the regulation of inflammatory cell accumulation in the extravascular periodontal connective tissues via cytokine production and surface expression of adhesion molecules. In this study, it was investigated the molecular basis for the adhesive interactions between monocytes and fibroblasts such as peri-odontal ligament fibroblast(PDLF), human gingival fibroblast(HGF), and human dermal fibroblast(HDF). First, it was examined the evidence whether monocyte-fibroblast cell contact may cause signal transduction in fibroblasts. Being directly in contact with fixed human monocyte cell line THP-1, or U937, upregulation of IL-6 production, $TNF-{\alpha}$ mRNA expression and increased cell proliferation could be seen for fibroblasts. IL-6 production induced by monocyte- fibroblast coculture were further increased when fibroblasts had been pretreated with $IFN-{\gamma}$ or $IL-1{\beta}$ , and monocytes with LPS. Next, it was examined the expression of ICAM-1 which has been known to be involved in accumulation and activation of leukocytes in inflammatory diseases such as periodontitis. ICAM-1 was upregulated up to 10-fold on PDLF, HGF, and HDF by exposure to $IFN-{\gamma}$ or $IL-1{\beta}$. Furthermore, anti-ICAM-1 monoclonal antibody clearly blocked cocultureinduced IL-6 production by fibroblasts, suggesting that $ICAM-1/{\beta}_2$integrin pathway is involved in periodontal fibroblastmonocyte interaction. Overall, these findings provide evidence that periodontal fibroblasts could be involved in the accumulation and retention of monocytes/macrophages in periodontal inflammatory lesion at least in part by ICAM-1 expression. In addition, periodontal fibroblast-monocyte interaction could cause activation signals in fibroblasts intracellularly which result in cytokine production and cell proliferation. Thus, periodontal fibroblasts are speculated to play an important role in immunoregulation and tissue destruction in chronic periodontal diseases by interaction with monocytes/macrophages.

• Journal of Periodontal and Implant Science
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• v.35 no.1
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• pp.163-176
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• 2005

3개월 동안 사용한 마모된 칫솔의 마모 정도와 양상을 관찰하고, 새 칫솔과 마모된 칫솔의 잇솔질 전 ${\cdot}$ 후 치태제거효율을 single-use design으로 비교 ${\cdot}$ 평가하여 3개월 주기의 칫솔 교체 주기의 근거를 임상적으로 확인 해보고자 하였다. 치주적으로 건강한 치과 대학생 42명을 대상으로 설문지를 통해 잇솔질 습관을 조사하고, 3개월간 동일한 칫솔과 치약을 사용하게 하였다. 3개월 후 피시험자를 무작위로 두 군(I, II)으로 나누고, 치석제거술을 시행한 뒤 2주후에 내원하도록 하였으며 내원 전 48시간동안은 잇솔질을 하지 않도록 지시하였다. 2주후 I군은 새 칫솔을, II군은 마모된 칫솔을 사용하도록 하였으며 잇솔질 전 ${\cdot}$ 후에 각각 구강 내를 erythrosin으로 염색한 후 6개의 Ramfjord 치아의 plaque score를 Patient Hygiene Performance (PHP) index로 측정하였다. 2주간의 washout period 후에 다시 치석제거술을 시행한 뒤, I군이 마모된 칫솔을, II군은 새 칫솔을 사용하게 하여 동일한 방법으로 PHP index를 각각 측정하였다. 마모된 칫솔은 수거하여 brushing surface area의 면적으로 마모도를 평가하였다. 결과는 paired t-test와 Pearson's correlation analysis로 통계처리 하였다. 2명이 탈락하였고 잇솔질 전 ${\cdot}$ 후에 대한 전체 부위, 치간 부위, 변연치은 부위의 plaque score는 두 칫솔 모두 통계학적으로 유의성 있게 감소하였으며 (p<0.0001), 두 칫솔을 비교한 경우에는 새 칫솔이 마모된 칫솔보다 치태 감소량이 통계학적으로 유의성 있게 많았다 (p<0.0001). 칫솔의 마모도는 평균 50.6% 증가하였으며, 마모도 증가에 따른 치태 감소량에는 직선적인 상관관계가 있었으나 통계학적인 유의성은 없었다. (전체 부위 r=-0.58, p=0.72 / 변연치은 부위 r=-0.50, p=0.76). Single-use design에서 3개월 동안 마모된 칫솔은 치태제거 능력에 있어서 새 칫솔보다 덜 효율적이였다. 칫솔의 마모도는 구강 위생 관리에 영향을 미치는 중요한 요인이며, 마모된 칫솔은 정기적인 교체가 요구된다. 또한, 치간 부위를 포함한 변연치은 부위의 치태를 정확하게 평가할 수 있는 치태지수에 대한 연구가 필요하겠다.

• Journal of Periodontal and Implant Science
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• v.35 no.1
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• pp.153-161
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• 2005

파이브로넥틴은 세포외기질에 존재하는 당단백질로 세포의 부착, 이동, 성장 및 분화에 관여하며, 섬유아세포 성장인자는 세포의 증식 이동 및 분화에 영향을 주는 중요한 성장인자로 알려져 있다. 최근 연구에 의하면, 파이브로넥틴은 조골세포의 타이태늄 임플란트 표면으로 이주와 증식 및 골생성을 촉진하며, 섬유아세포 성장 인자는 파이브로넥틴에 상승작용을 한다고 보고된 바 있다. 이 실험의 목적은 파이브로넥틴 및 섬유아세포 성장인자의 복합 단백질을 이용하여 타이태늄 임플란트의 골 반응을 알아보는 것이다. 체중 2.5 kg 내외의 건강한 18 마리의 웅성가토를 준비하여 무균 사육하였고, 순수 타이태늄을 절삭가공하여 직경 3.5mm, 길이 6mm 의 machined surface를 지니는 screw type 의 임플란트를 준비하였다. 사람의 유전자를 기초로, 유전자 재조합법을 통해, 적절한 primer를 이용하여 얻은 섬유아세포 성장인자를 파이브로넥틴 III 형 분절의 9-10 번 도메인에 결합시켜 얻은 복합 단백질을 준비된 임플란트에 표면처리하여 실험군으로 하였고, 표면처리하지 않은 임플란트를 대조군으로 하여, 가토의 좌우 경골에 각각 2 개씩의 임플란트를 식립하였다. 4주 후, 가토를 희생시켜 각 경골 당 한 개의 임플란트에서 뒤틀림 제거력을 측정하였고 나머지 임플란트 식립 부위 에서는 경골을 포함하는 조직표본을 제작하였다. 조직표본상에서 골접촉이 가장 좋은 3 개의 나사산의 길이를 측정하고, 나사와 접촉하는 골의 길이를 측정하여 골-임플란트 접촉도를 구하고, 같은 부위에서 나사산 사이의 면적과 골이 차지하는 면적을 비교하여 골생성률을 얻었다. 실험군과 대조군의 결과는 Student t-test 를 이용하여 신뢰도 95% 수준에서 통계학적 유의성을 검정하였다. 파이브로넥틴과 섬유아세포 성장인자의 복합 단백질로 표면처리된 임플란트와 표면처리를 하지 않은 임플란트는 뒤틀림 제거력에서는 통계적 유의성이 나타나지 않았으나, 골-임플란트 접촉도와 골생성률에서 복합 단백질로 처리된 임플란트가 통계적으로 유의하게 높은 결과를 보였다. 이상의 연구결과로, 섬유아세포 성장인자와 파이브로넥틴 복합 단백질로 처리한 타이태늄 임플란트가 주변 골 형성을 촉진시켜, 골유합을 증진시킴을 알 수 있었다. 따라서, 복합 단백질이 타이태늄 임플란트의 성공률을 높이기 위한 표면개질 물질로 이용될 가능성을 확인할 수 있었다.