• Journal of Periodontal and Implant Science
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• 제25권3호
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• pp.497-517
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• 1995

The cells associated with normal defense mechanism in inflammation release free oxygen radicals, hydroxy radicals, and various protease, all of which can damage the surrounding cells(fibroblasts) and matrix molecules(collagen). The objective of this study was to evaluate the effects of "scavenger" enzyme, superoxide dismutase(SOD). to periodontal ligament (PDL) cells. Human PDL cells were cultured from the teeth extracted for non-periodontal reason. Cultured PDL cells in vitro were treated with SOD and LPS according to dosage and culture times. Cellular activity was exaimed by Microtitration(MTT) assay. The quantitative expression of cellular proliferation by proliferating cell nuclear antigen(PCNA), collagen type I and fibronectin by indirect immunocytochemically stain in PDL cells were done. The results were as follows: 1. As only SOD treated group at 2 and 3 days, PDL cell activity was significantly increased at more than 150U(P<0.05). 2. When LPS(0.5, $5{\mu}g/m{\ell}$) and SOD(more than 150U) were added together, it was significantly increased than LPS only treated and control groups at 2 days(P<0.05). 3. When LPS($5{\mu}g/m{\ell}$) and SOD(150, 300U) were added together, PCNA index was significantly increased than LPS only treated and control groups at 2 and 3 days(P<0.05). 4. When LPS($5{\mu}g/m{\ell}$) and SOD(150U) were added together, collagen type I was significantly increased than LPS only treated and control groups at 3 days(P<0.05). 5.When LPS($5{\mu}g/m{\ell}$) and SOD(300U) were added together, fibronectin was significantly increased than LPS only treated and control groups at 3 days(P<0.05). On the above the results, the SOD in association with collagen type I, fibonectin, and PCNA may afford biological protection to oxy-radicals that were typically liberated during normal inflammatory response. Thus, the exogenous application of SOD may be effective in sthe treatment of the localized breakdown associated with chronic periodontal disease.

• Journal of Periodontal and Implant Science
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• 제32권3호
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• pp.445-456
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• 2002

치주조직 재생을 위해서는 새로운 백악질과 치조골 그리고, 치주인대의 재생이 필요하며, 이러한 재생을 담당할 세포의 분화가 필수적이다. 이러한 분화를 담당하는 것은 치주인대세포이며, 이 중 골아세포의 분화가 중요하다. 본 실험의 목적은 치주조직 재생에 있어서 중요한 요소인 치주인대세포의 골아세포성 세포로의 분화를 관찰하며, Dex가 광물화에 미치는 영향과 농도에 따른 차이를 알아보고자 시행하였다. 또한, 광물화시 발현되는 여러 골기질 단백질 중 Matrix GlaProtein의 발현양상도 관찰하였다. 교정치료를 목적으로 내원한 환자의 제1소구치 부위의 정상치은을 절제하고, 건강한 제1소구치를 발거하여 치은섬유아세포와 치주인대세포를 분리, 배양하여, ascorbic acid와 ${\beta}$-glycerophosphate 투여군을 실험1군, ascorbic acid, ${\beta}$-glycerophosphate, Dex 100nM 투여군을 실험 2군, ascorbic acid, ${\beta}$-glycerophosphate, Dex $5{\mu}M$ 투여군을 실험3군, 그리고, 단순 배양만 시킨군을 대조군으로 하여 비교하였다. 시간경과에 따른 치주인대세포 형태의 변화 양상은 초기에 방추형 혹은 다각형의 단일층 형태에서 7일경에는 세포 크기와 수가 증가하여 복합층 형태로 변화했으며, 배양 14일 이후에는 세포들의 방향성이 없어지고, 더욱 치밀해 졌다. 골 결절형성은 치주인 대세포의 Dex 투여군에서만 21일째에 나타났으며, $5{\mu}M$ 투여군에서 100nM 투여군보다 더 많이 나타났다. ALP 활성도를 비교해보면 치주인대세포에서 0, 7일 경에는 활성도를 보이지 않았으며, 14일경에 높은 활성도롤 나타냈으며, 21일에도 비슷한 활성도를 유지하였다. MGP 유전자 발현 양상은 대조군과 실험군 모두에서 Matrix Gla Protein에 대한 유전자의 발현이 나타났으며，그 발현양상은 모든 시기에서 일정하였다. 이상의 결과로 보아 치주인대세포는 골아세포로의 분화가 가능하며, Dex는 농도의존적으로 광물화에 영향을 미치는 것으로 사료된다. 그리고, MGP는 치주인대세포에서 발현이 감지되었으며, 광물화에는 영향을 미치지 않는 것으로 사료된다.

• 대한치과교정학회지
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• 제36권4호
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• pp.242-250
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• 2006

기계적 응력은 정상적인 발달과정 동안 조직의 항상성에 있어 중요한 역할을 한다. 기계적 응력은 치아 이동과 저작과 같은 상황을 포함한다. 치아 이동과 저작 중에 치주인대 섬유모세포는 기계적 자극을 감지하고 주위의 세포밖 물질과 생체분자 대사의 변동에 의해 반응을 보인다. 그러나 아직까지 기계적 응력 하에서 치주인대 세포에서 발현된 유전자에 관한 연구는 미비한 실정이다. 최근 기계적 응력이 초파리에 존재하는 UNC-50 유전자에 영향을 줄 수 있다고 보고되었다. 또한 UNC-50은 치은 섬유모세포와 비교해 치주인대 섬유모세포에서만 발현된다고 보고되었다. 본 연구에서는 간헐적 교정력 적용 시에 백서의 치아에서 일어나는 치근 및 치주 조직의 조직학적 변화와, UNC-50의 발현 양상을 면역조직학적 염색으로 조사하여 치주인대에서 기계적 응력과 UNC-50의 관련성을 알아보고자 하였다. Sprague-Dawely계 수컷 백서 12마리를 4마리씩 세 군으로 나누어 상악 우측 구치부에 NiTi closed coil spring을 사용하여 40 g 정도의 견인력이 발생하도록 하여 하루에 1시간씩 간헐적인 교정력을 적용한 후 1, 3, 5일 후 치주인대의 조직학적 변화를 관찰하여 다음과 같은 결과를 얻었다. 조직학적 소견에서 상악 제1대구치 근심구개치근의 치근부 1/3에서 압박측은 안장측보다 더 좁은 치주인대공간을 보였고 교정력을 적용시킨 후 3일 후부터 인장측에서 백악모세포 활성으로 인한 백악질 침착이 관찰되었다. UNC-50은 인장측의 분화 중인 백악모세포에서 강한 발현을 보였다. Osteocalcin은 인장측에서 압박측에 비해 신생 백악질에 존재하는 분화 중인 백악모세포를 따라 강한 발현을 보였다. 이상의 연구결과는 UNC-50이 간헐적 교정력 즉 기계적 응력의 변화에 따른 백악모세포의 분화과정에 중요한 역할을 함을 나타낸다. 그러나 이를 명확히 하기 위해서는 교정력 적용 후 UNC-50의 세포내 신호전달과정에 대한 보완연구가 필요할 것이다.

• 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.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제34권5호
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• pp.532-536
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• 2008

Purpose: This study was designed to evaluate effect of EMD on proliferation of HPDLCs and MC3T3-E1 cells in high glucose condition in vitro. Material and method: The Human PDL fibroblasts(HPDLCs) were obtained through typical way and the cells used in this experiment were divided in 4 groups. $1{\times}10^4/ml$ HPDLCs suspension was cultured in typical DMEM and assigned to group 1. The cells cultured in DMEM which included 400mg/dl glucose are allocated to group 3. Group 2 and 4 are established by adding EMD to group 1 and 3 respectively. These control and experimental groups had been cultured for 24 and 48 hours, and MTT assay was conducted. The differences of each group in cellular proliferation was evaluated. The same experiment was conducted for preosteoblast (MC3T3-E1) with adding $25\;{\mu}g/ml$ EMD. Results: EMD had the same effect on both PDL cells and MCT3T3-E1 cells. The experimental group had more meaningful differences and active cellular proliferation than the control group did. The EMD accelerated cellular proliferation not only in normal glucose condition but also in high glucose condition. The same results were observed via MTT assay; EMD-added experimental group had more meaningful differences and showed higher cellular activity than control group did. Each experimental and control group was inspected for statistical significance through Kruskal-Wallis Test. Statistical significances were observed among these groups. (SPSS 12.0 Chicago, IL, USA, p=0.008, p=0.011) Conclusion: EMD is considered to accelerate proliferation of PDL cells and MC3T3-E1 cells in high glucose condition as well as normal glucose condition.

• Journal of Periodontal and Implant Science
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• 제25권2호
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• pp.279-289
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• 1995

The most important object of periodontal treatment is the perfect regeneration of destructed periodontal tissue. The healing of periodontal lesion is affected by several cells & factors, which result in formation of long juntional epithelium, root resorption, bony ankylosis or connective tissue attachment. And ideal healing is enhanced by epithilial exclusion or periodontal ligament cell activation. In this investigation, I studied the effect of Zizyphus Fructus extract which enhances biologic activity& collagen synthesis, on the chemotaxis & cell nature. The cells were obtained from interdental area & middle third area of the freshly extracted teeth for the orthodontic purpose. And they were fully incubated in${\alpha}-MEM$ solution containing $100{\mu]g/ml$ penicillin & $100{\mu]g/ml$ streptomycin followed by 6 generation incubation. The test cells were collected by trypsin-EDTA & centrifuge in the fully incubated cells, counted by Hernacyotmeter, incbated $5{\times}10^5/ml$ cells for 24 hours, re-incubated 24 hours in media containing natural extract and photographed. The cells were incubated for 4 hours in 48 well microchemotaxis chamber bisecting upper & lower chamber by 8ug/m pore polycarbonate membrane coating 5mg/ml gelatin solution. The migrated cells in microscope were counted, which meaned cell chemotaxis activity. The study had shown that the morphology of cell was spindle-shaped as the control group, and the subextract test groups were not significantly different. In gingival fibroblasts, the chemotaxis effect of PDGF was statistically significant compared to control group. The Zizyphus Fructus extract was more or less enhanced chemotaxis effect and in $1{\mu}g/ml$ concentration the chemotaxis effect was slightly elevated compared with $10{\mu}g/ml$ concentration. But, among the subextracts, it was not significantly defferent. In PDL cells, the chemotaxis effect of PDGF in statistically significant, and the zizyphus Fructus extract had shown the enhanced effect. The effect was slightly higher in $1{\mu}g/ml$ concentration than 10g/ml concentration,and no significance among the subextracts.