• 대한치과교정학회지
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• 제31권4호
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• pp.447-458
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• 2001

치아이동시 골대사에 있어 glycosaminoglycan의 역할에 대해 알아보고자 glycosaminoglycan의 주요 구성 성분인 chondroitin 4-sulfate(CH-4S)의 치주조직 내에서의 면역반응 정도 및 분포 양상을 백서 치아의 실험적 이동 과정에서 면역조직화학적으로 관찰하였다. 또한 사람 치주인대세포를 배양하여 여러 종류의 cytokine을 투여한 후 CH-4S의 발현 양상의 변화를 Western blot analysis를 통해 확인하여 다음과 같은 결과를 얻었다. 1. 치아이동에 반응하는 치수, 치주인대, 골모세포, 파골세포, 골세포 부위에서의 CH-4S 발현이 대조군보다 많았으나 상아질, 백악질에서의 CH-4S의 발현은 견인력 적용 기간에 관계없이 대조군과 큰 차이가 없었다. 2. 치수에서 CH-4S의 발현은 교정력을 가한 1일째에 크게 증가하였다가 7일째부터 감소되었으며 14일째에는 대조군과 차이가 없었다. 3. 치주인대에서 CH-4S의 발현은 주로 치조골 면을 따라 견인측에서 나타났는데 교정력을 가한 1일째에 가장 많은 발현을 보인 후 4일째부터 감소하기 시작하였다. 4. 골모세포와 파골세포 및 골세포에서 CH-4S의 발현은 4일째에 가장 많은 발현을 보였고 7일째 이후에는 크게 감소하였다. 3. 치주인대세포에 PDGF-BB를 투여한 경우 3일째에 가장 많은 CH-4S의 발현을 보였다. 6. 치주인대세포에 $TNF-\alpha$ 처리 시 배양 1일째에 CH-4S의 발현 감소를 보였다. 7. 치주인대세포에 PDGF-BB와 $TGF-\beta$를 혼합 투여 한 경우가 PDGF-BB 및 $TGF-\beta$를 단독 투여한 경우 보다 배양3일째에 CH-4S의 발현이 많았고 LPS나 $TNF-\alpha$ 투여군은 유사한 발현 감소를 보였다. 이상과 같이 교정적 치아이동시 시기 및 부위에 따라 glycosaminoglycan의 발현이 차이를 보이며, 치주인대세포에서도 cytokine의 자극에 따라 glycosaminoglycan의 발현이 변화하는 것으로 보아, glycosaminoglycan이 골대사에 있어 중요한 조절 인자로서의 역할을 하는 것으로 여겨진다.

• Journal of Periodontal and Implant Science
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• 제23권1호
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• pp.37-47
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• 1993

The native connective tissue attachment of the periodontium is known to be a complex consisting of gingival fibroblasts, periodontal ligament cells, gingival epithelial cells, cementum, alveolar bone and extensive extracellular matrix (collagen, glycoprotein and proteoglycans). The purpose of this study was to evaluate the effects of natural extracts on DNA, collagen and protein synthesis and inhibition of cytokine production in the gingival and periodontal ligament fibroblasts and gingival epithelial cells. Healthy gingival tissue was obtained from orthodontic treatment patients, and gingival epithelial cells, gingival fibroblasts and periodontal ligament cells were isolated and cultured from the samples. After treated with Ginseng protein, Pluronic F-68, Scutellariae Radix, centella asiatica, PDGF, IGF, DNA synthesis, total protein and collagen synthesis, and cytokine production of gingival epithelial cell, gingival fibroblast and periodontal ligamentcells were measured. MTT method for DNA synthesis, Peterkofsky and Dingerman method for total protein and collagen synthesis, and IL-1 ELISA kit for cytokine production were used. The proliferation of epithelial cells was enhanced in Centella asiatica, Ginseng protein, Pluronic F-68 and Scutellariae Radix. The activities of PDL cells were increased in PDGF, IGF, and Pluronic F-68. Higher collagen synthesis was observed in Scutellariae Radix and total protein synthesis was increased in Scutellariae Radix and PDGF. The inhibitory effects on IL-1, IL-6, $TNF-{\alpha}$ were observed in all exrracts.

• BMB Reports
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• 제42권7호
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• pp.427-432
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• 2009

Orthodontic tooth movement results from the combinational process of both bone resorption and formation in the compressive and tension sides, respectively. However, the genes responsible for new bone formation in tension sides have not been determined. In this study, we used DNA microarray and real-time RT-PCR to identify genes in human periodontal ligament (PDL) cells that undergo significant changes in expression in response to static tensional forces (2 or 12 hours). The genes found were alkaline phospatase (ALP), matrix metalloproteinases (MMPs), vascular endothelial growth factor (VEGF), and several collagen genes. Furthermore, an ELISA evaluating the expression of VEGF, type IV collagen and MMP-2 found levels significantly increased after 24 and 72 hours (P < 0.05). ALP activity was also increased after 24 hours (P < 0.05). Collectively, we found the genes up-regulated in our study by the static tensional force are related to osteogenic processes such as matrix synthesis and angiogenesis.

• Restorative Dentistry and Endodontics
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• 제43권3호
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• pp.24.1-24.12
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• 2018

Objectives: The aim of this in vitro study was to evaluate the biocompatibility of newly proposed root-end filling materials, Biodentine, Micro-Mega mineral trioxide aggregate (MM-MTA), polymethylmethacrylate (PMMA) bone cement, and Smart Dentin Replacement (SDR), in comparison with contemporary root-end filling materials, intermediate restorative material (IRM), Dyract compomer, ProRoot MTA (PMTA), and Vitrebond, using human periodontal ligament (hPDL) fibroblasts. Materials and Methods: Ten discs from each material were fabricated in sterile Teflon molds and 24-hour eluates were obtained from each root-end filling material in cell culture media after 1- or 3-day setting. hPDL fibroblasts were plated at a density of $5{\times}10^3/well$, and were incubated for 24 hours with 1:1, 1:2, 1:4, and 1:8 dilutions of eluates. Cell viability was evaluated by XTT assay. Data was statistically analysed. Apoptotic/necrotic activity of PDL cells exposed to material eluates was established by flow cytometry. Results: The Vitrebond and IRM were significantly more cytotoxic than the other root-end filling materials (p < 0.05). Those cells exposed to the Biodentine and Dyract compomer eluates showed the highest survival rates (p < 0.05), while the PMTA, MM-MTA, SDR, and PMMA groups exhibited similar cell viabilities. Three-day samples were more cytotoxic than 1-day samples (p < 0.05). Eluates from the cements at 1:1 dilution were significantly more cytotoxic (p < 0.05). Vitrebond induced cell necrosis as indicated by flow cytometry. Conclusions: This in vitro study demonstrated that Biodentine and Compomer were more biocompatible than the other root-end filling materials. Vitrebond eluate caused necrotic cell death.

• 대한치과교정학회지
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• 제30권4호
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• pp.441-452
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• 2000

본 실험은 사람의 치주인대세포에서 에스트로겐과 growth hormone(GH)이 상호 어떠한 작용을 하는지를 규명하는 것이 목적이다. 교정치료를 받고자 내원한 환자중에서 건강한 20대 여자환자들의 상하악 제1소구치를 발거하여 치근의 중간1/3부위에서 긁어모은 치주인대 조직을 배양하여 치주인대세포를 얻었다. 사람의 치주인대세포의 분열증식에 대한 $17{\beta}$-estradiol과 hGH의 효과를 평가하고, 치주인대세포에 $17{\beta}$-estradiol을 전처리한 후 치주인대세포의 분열증식에 미치는 hGH의 효과 변화를 평가하였으며, 치주인대세포에 $17{\beta}$-estradiol을 전처리 하였을 경우 치주인대세포에 서 hGH receptor치 발현 양상의 변화를 평가하여 다음과 같은 결론을 얻었다. 1. $17{\beta}$-estradiol이나 humangrowth hormone의 단독처리는 사람의 치주인대세포의 분열종식에 큰 영향을 주지 않는다. 2. $17{\beta}$-estradiol $10^{-12}M$로 전처리한 후 hGH를 투여한 경우 hGH의 농도에 관계없이 사람의 치주인대세포의 분열증식을 증가시킨다. 3. 사람의 치주인대세포에는 hGH receptor가 없으나, $17{\beta}$-estradiol $10^{-12}M$로 5시간 이상 처리하면 hGH receptor가 발현된다. 4. hGH이 사람의 치주인대세포의 분열증식에 미치는 효과는 hGH receptor의 발현과 관련이 있으며 $17{\beta}$-estradiol의 전처리가 치주인대세포에서의 hGH receptor의 발현에 기여함으로써 hGH이 치주인대세포에 작용할 수 있도륵 해준다.비, 상악의 대구치간 너비, 구개정 높이가 상관관계가 있었다.에서 치 아이동은 정상상태보다 촉진될 가능성이 있음을 시사한다.따라 평균마찰력은 유의하게 증가하였다. 6. 브라켓간 거리의 변화에 대한 마찰력의 차이는 교정선의 재질에 따라 차이가 나며, 스테인레스 스틸 교정선은 브라켓간 거리가 감소할 때 마찰력이 유의하게 변화하지 않았으나, NiTi 교정선의 경우는 유의하게 증가하였다. 7. 브라켓내의 교정선의 이동속도에 따라 마찰력은 유의하게 변화하지 않았다. 이상의 결과로 볼 때 교정 치료동안 적정 교정력을 유지하기 위해 자가결찰브라켓, 스테인레스 스틸 교정선과 탄성모듈결찰법을 사용하는 것이 유리하며, 치료시기에 따라서 요구하는 마찰력이 다르므로 상황에 따라 재료를 선택하는 것이 중요할 것으로 사료된다.$, B군에서 $4.3\%$ 감소하였다. 7. 수술로 인한 얼굴폭의 변화는 무시할만한 것으로 판단되나 수술후 얼굴의 길이가 짧아져 상대적으로 얼굴의 폭이 넓어보일 수 있으므로 수술시 이에 대한 충분한 고려 및 환자에 대한 설명이 있어야 한다.로는 4-6세군 ($27.5\%$), 6-8세군 ($19.6\%$), 2-4세군 ($13.7\%$)이 $60\%$이상을 차지하여 초등학교 취학 전에 구순의 반흔을 제거하려 함을 알 수 있었다. 8. 비변형 교정수술시기로는 0-2세군 ($7.1\%$), 2-4세군 ($14.3\%$), 4-6세군 ($21.4\%$), 6-8세군 ($14.3\%$)으로 초등학교 취학이전이 $57.1\%$로서 최근의 조기 치료경향을 반영하는 것으로 보인다. 9. 인두피판술은 평균 6세에 시행되었으며, 수술 시행 시기별의 차이를 보이지 않고 고른 분포를

• 대한치과교정학회지
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• 제24권4호
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• pp.871-883
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• 1994

This experiment was designed to study possible roles of $interleukin-1\beta$, interleukin-6 and tumor necrosis $factor-\alpha$ in bone remodeling by measuring their effects on $PGE_2,\; LTB_4$ and collagenase production when they were administered to human periodontal ligament fibroblasts. Human periodontal ligament fibroblasts were collected from first premolars extracted for orthodontic treatment. They were incubated in the environment of $37^{\circ}C,\;5\%\;Co^2,\;and\;100\%$ humidity. They were treated with $0.25\%$ trypsin-EDTA solution and centrifuged. PDL cells in the fifth to seventh passage were used for the experiment. Cells were seeded onto the culture dishes and when they were successfully attached, human recombinant $interleukin-1\beta$, interleukin-6, and tumor necrosis $factor-\alpha$ were administered, alone or in combination. They were incubated for 4, 8 and 24 hours and the levels of $PGE_2,\;LTB_4$ and collagenase released into the culture media were assessed by enzymeimmunoassay and collagenase activity assay. The conclusions are as follows: 1. $IL-1\beta\;and\;TNF-\alpha$ were very active in stimulating the production of $PGE_2$ and collagenase by human periodontal ligament fibroblasts, while IL-6 increased $LTB_4$ production. 2. $IL-1\beta$ significantly increased $PGE_2$, but $LTB_4$ Production was not increased. $IL-1\beta$ is thought to act mainly via the cyclooxygenase pathway of arachidonic acid metabolism. 3. IL-6 tended to inhibit $IL-1\beta$ in the production of $PGE_2$ and collagense whereas IL-6 and $TNF-\alpha$ showed auditive effect in the level of $PGE_2$. The above cytokines increased the release of at least one of $PGE_2,\;LTB_4$ and collagenase. It suggests that cytokines are involved in bone remodeling process by stimulating PDL fibroblasts to produce various bone-resorptive agents. The roles of cytokines in bone remodeling as a whole would need further study.

• Journal of Periodontal and Implant Science
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• 제32권1호
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• pp.1-12
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• 2002

The periodontal ligament(PDL) is a unique tissue that is crucial for tooth function. However, little is known of the molecular mechanisms controlling PDL function. PDL-specific protein;PDLs22 had been previously identified as a novel protein isolated from cultured human PDL fibroblasts using subtraction hybridization between human gingival fibroblasts and PDL fibroblasts. The aim of this study was to examine the expression pattern and tissue localization of PDLs22 protein in embryonic and various postnatal stages of developing mouse using immunohistochemical staining. Embryos (E18) and postnatal (P1, P4, P5, P15, P18) were decapitated and the heads were fixed overnight in a freshly prepared solution of 4% paraformaldehyde. Some specimens were decalcified for $2{\sim}4$ weeks in a solution containing 10% of the disodium salt of ethylenediamine-tetraacetic acid (EDTA). Next, tissues were dehydrated, embedded in paraffin and sectioned serially at $6{\mu}m$ in thickness. Polyclonal antiserum raised against PDLs22 peptides, ISNKYLVKRQSRD, were made. The localization of PDLs22 in tissues was detected by polyclonal antibody against PDLs22 by means of immunohistochemical staining. The results were as follows; 1. Expression of PDLs22 protein was not detected in the tooth germ of bud and cap stage. 2. At the late bell stage and root formation stage, strong expression of PDLs22 protein was observed in developing tooth follicle, osteoblast-like cells, and subodontoblastic cells in the tooth pulp, but not in gingival fibroblasts, ameloblasts and odontoblasts of tooth germ 3. In erupted tooth, PDLs22 protein was intensely expressed in PDL and osteoblast-like cells of alveolar bone, but not in gingival fibroblasts, mature osteocytes and adjacent salivary glands. 4. In the developing alveolar bone and mid-palatal suture, expression of PDLs22 protein was seen in undifferentiated mesenchymal cells and osteoblast-like cells of developing mid-palatal suture, but not in mature osteocytes and chondrocytes. These results suggest that PDLs22 protein may play an important role in the differentiation of undifferentiated mesenchymal cells in the bone marrow and PDL cells, which can differentiate into multiple cell types including osteoblasts, cementoblasts, and PDL fibroblasts. However, more researches should be performed to gain a better understanding of the exact function of PDLs22 protein which related to the PDL cell differentiation.

• 대한치과교정학회지
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• 제28권1호
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• pp.143-153
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• 1998

PDGF와 LPS는 골 대사에 중요한 조절요소이다. 외인성 PDGF는 골 흡수를 자극하기도 하고, 또 새로운 골 형성을 자극하기도 한다. LPS는 골파괴 활성의 자극 요인으로 알려져 있다. 본 연구의 목적은 골개조에 중요한 역할을 하는 치주 인대에 PDGF와 LPS의 상호 작용을 평가하기 위해서 시행되었다. 배양된 사람의 치주 인대 세포에 PDGF와 LPS를 첨가 배양하여 각 농도와 시간에 따른 대조군에 대한 백분율로 환산된 세포 활성의 평균과 표준 편차를 구하고 ANOVA를 시행하여 다음과 같은 결과를 얻었다. 1. PDGF를 단독으로 가한 경우 0.1ng/ml에서는 세포 활성이 대조군과 차이가 없었으나 1ng/ml에서는 배양 2일째에, 10ng/ml에서는 배양 2,3일째에 유의한 세포 활성 증가를 보였다. 2. LPS단독처리시 $0.5{\mu}g/ml$ 및 $5{\mu}g/ml$농도에서 배양 1일째,2일째에는 대조군과 차이가 없었으나 배양 3일째에 유의한 활성 감소가 없었다. 3. $0.5{\mu}g/ml$ 및 $5{\mu}g/ml$농토의 LPS에 1ng/m 및 10ng/ml농도의 PDGF 첨가시, 배양 3일째에 LPS단독 투여시보다 유의한 세포 활성 증가가 있었으며 특히 10ng/ml PDGF 첨가시에는 대조군보다 활성이 컸다. 이상의 연구결과로 골개조 과정에 관여하는 것으로 여겨지는 치주 인대세포에 대한 적절한 농도의 PDGF와 LPS의 복합 투여는 PDGF 단독 투여 못지 않게 치주 인대세포의 활성을 증가시키는 것으로 사료된다.

• Journal of Periodontal and Implant Science
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• 제28권1호
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• pp.103-120
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• 1998

Prokaryotic and eukaryotic cells respond to heat stress and other environmental abuses by synthesizing a small set of stress proteins and by inhibiting post-transcription synthesis of normal proteins. The purpose of the present study was to document the stress response produced by inflamed gingival tissue in vivo, and cytokine inducted human periodontal ligament cells. Human PDL cells were exposed to TNF-$\alpha$(1ng/ml), INF-$\gamma$(200 U/ml), LPS(100ug/ml), combination of cytokine, and SDS-PAGE gels running and Western blotting analysis was done. In vivo studies, the healthy gingival tissusse of a control group and inflamed gingival tissue of adult periodontitis were studied by immunohistochemistry and histology. The results were as follows 1. HSP 47 was distributed on basal layer in healthy gingiva, but stronger stained in basal, suprabasal, and spinous layer of inflamed gingiva. 2. HSP 47 was rare on endothelial cells and mononuclear cells in healthy gingiva, but stronger expressed in inflamed gingiva. 3. HSP 70 expression was rare on epihelium and inflammatory cells hi both healthy & inflamed gingiva. 4. HSP 70 was actively expressed on endothelial cells and inflammatory cells of capillary lumen in moderately & mild inflamend gingiva. 5. PDL cells showed low level of HSP 47 protein expression which was significantly induced by cytokine stimulation (LSP only and combination). 6. Maximum HSP 70 protein induction was seen with stimulation by a combination of the cytokine, Combination of TNF-$\alpha$, INF-$\gamma$, LPS have been shown to synergistically effects of HSP 70 expression. On the above findings, HSP Is influenced by cytokine and chronic inflammation in vivo, and may be involved in protection of tissue during periodontal inflammatiom.

• 대한치과교정학회지
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• 제16권1호
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• pp.51-70
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• 1986

Parathyroid hormone (PTH) is known to exert its effects on bone cells through the mediation of adenosine 3', 5'-monophosphate (cAMP). Orthodontic forces have also been shown to alter the cAMP content of paradental cells, particularly the alveolar bone osteoblasts. The objective of this experiment was to determine whether a combined orthodontic treatment-PTH administration regimen would have an additive effect on cAMP content in paradental cells in sites of periodontal ligament (PDL) tension. Seven groups of 4 one year old female cats each were treated for 1,3,6,12,24 h, 7 and 14 d by tipping one maxillary canine. PTH was administered twice daily, 30u/kg. Maxillary horizontal sections were stained immunohistochemically for cAMP and the degree of cellular staining intensity was determined microphotometrically as per cent light transmittance at 600nm. Alveolar bone osteoblasts, progenitor cells, PDL fibroblasts and cementoblasts in tenion sites were measured and the data were analyzed statistically by a mixed model analysis of variance. PTH administration increased the cAMP staining of nonorthodontically treated paradental cells in comparison to cells untreated by force or hormone. Cells in PDL tension sites of PTH-treated cats demonstrated significantly darker cAMP staining than cells in non-orthodontically-treated sites. Osteoblasts demonstrated the greatest response in terms of cAMP elevation, while in PDL fibroblasts orthodontic force did not increase cAMP levels above those measured in non-stretched hormonally-treated cells. These results demonstrate that PTH increases cAMP levels in paradental cells, particullarly in osteoblasts, and that the effects of PTH and orthodontic forces on paradental target cells may approach additivity.