• Journal of Periodontal and Implant Science
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• v.26 no.1
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• pp.176-187
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• 1996

The ability of fibroblasts attached to teeth is paramount important in reestablishing the lost connective tissue attachment after periodontal therapy. The migration and proliferation of periodontal ligament cells are desired goal of periodontal regeneration therapy. PDGF is well known to regulate the cell activity of mesenchymal origin cell. Tobacco contains a complex mixture of substance including nicotine, various nitrosamines, trace elements, and variety of poorly characterized substances. Human gingival fibroblasts and periodontal ligament cells were cultured from extracted tooth for non-periodontal reason. Cultured human gingival fibroblasts and periodontal ligament cells in vitro were treated with PDGF, nicotine in time dependent manner. Cellular activities were determined by MTT assay. The purpose of this study was to determine the effects of Nicotine and PDGF, respectively and the effect of PDGF presence of nicotine on human gingival fibroblasts and periodontal ligament cells. The results were as follows : 1. In the cell activities of human gingival fibroblasts and periodontal ligament cells were similar or decreased to control value at 1st day. At 2nd day, cellular activities of both group were increased to control value. At 3rd day, cellular activities of both group were returned to the control value. 2. In the cell activities of PDGF on human gingival fibroblasts and periodontal ligament cells, cell activities significantly increase from control group on periodontal ligament cells compared to gingival fibroblast group at 3rd day. 3. In the cell activities of PDGF and nicotine combined application on human gingival fibroblasts and periodontal ligament cells, it seems likely that the nicotinic effect of gingival fibroblasts were higher than periodontal ligament cells and the PDGF effect of periodontal ligament cells were higher than gingival fibroblasts. This results suggested that PDGF might stimulate the selective growth on periodontal ligament cells.

• Journal of Periodontal and Implant Science
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• v.25 no.1
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• pp.133-145
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• 1995

The migration and proliferation of periodontal ligament cells are desired goal of periodontal regeneration therapy. PDGF and $TGF-{\beta}1$ are well known to regulate the cell activity of mesenchymal origin cell. The purpose of this study was to determine the effects of these growth factors on human gingival fibroblast and periodontal ligament cell actvity, and to identify the regulatory effect of $TGF-{\beta}1$ on the response to PDGF by MIT assay. Human gingival fibroblast and periodontal ligament cells were cultured from extracted teeth for non-periodontal reason. Cultured human gingival fibroblast and periodontal ligament cells in vitro were treated with polyperpetide growth factor PDGF and $TGF-{\beta}1$ in both a dose and time - dependent manner. Cell morphology were determined by inverted microscope and cell acitivity were determined by MIT assay. The result of this study demonstrated that PDGF and $TGF-{\beta}1$ were not changed the morphology of these cell compared with control group. PDGF or $TGF-{\beta}1$ increased cell activity of periodontal ligament cell in dose and time dependent manner but gingival fibroblast were decreased to the level of control group at third day. Additionally, incubation with $TGF-{\beta}1$ addition to PDGF resulted in a enhanced cell activity of PDGF. Therefore, cell acitivty of gingival fibroblast were not changed compared with control group. This stiudy demonstrates that PDGF and $TGF-{\beta}1$ are major mitogens for human periodontal ligament cell in vitro, and $TGF-{\beta}1$ is a regulator of cell activity to PDGF in human gingival fibroblast and periodontal ligament cell.

• Journal of Periodontal and Implant Science
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• v.31 no.1
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• pp.163-180
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• 2001

The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of periodontal ligament cells. Primary human periodontal ligament cells were cultured in dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells of 4th to 7th passage were inoculated in the multiwell plates coated with chitosan in concentration of 0.22, 0.2, and $2mg/m{\ell}$. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized modules was evaluated after 21 days of culture. The results were as follows : 1. The morphology of periodontal ligament cells on the chitosan coating was round or spheric. Round cells were aggregated after 6 hours of culture. Aggregated cells on the chitosan coated surface showed nodule-like appearance after 24 hours of culture and not achieved confluency at 7 days. 2. During early period of culture, the attachment of periodontal ligament cells were inhibited by chitosan coating. Inhibition of cell attachment tended to increase with the concentration of chitosan. 3. At the chitosan concentration of 0.02 and $0.2mg/m{\ell}$, periodontal ligament cells were more rapidly proliferated at 7 days, compared to the control group. At the concentration of $2mg/m{\ell}$, the proliferation of periodontal ligament cells was inhibitied(p<0.01). 4. Alkaline phosphatase activity of periodontal ligament cells was increased in chitosan coated group, especially at the concentration of $0.02mg/m{\ell}$after 4 days of culture.5. Periodontal ligament cells produced mineralized nodules on chitosan coated wells without the addition of mineralized nodule forming materials (ascorbic acid, ${\beta}-glycerophosphat$, dexamethasone). With the addition of mineralized nodule forming materials, periodontal ligament cells produced more mineralized nodules at the concentration of $0.02mg/m{\ell}$, compared to the control. In summary, the attachment, proliferation, cell activity, and alkaline phosphatase activity of periodontal ligament cells depended on the concentration of coated chitosan. Chitosan stimulated mineralized nodule formation by periodontal ligament cells. At the appropriate concentration($0.02mg/m{\ell}$), chitosan could increase alkaline phosphatase activity and stimulate the formation of mineralized nodule by periodontal ligament cells. These results suggest that chitosan can be used as an adjunct for bone graft material, and the matrix of tissue engineering for periodontal regeneration, especially bone regeneration.

• Journal of Periodontal and Implant Science
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• v.34 no.2
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• pp.367-375
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• 2004

As the periodontal ligament cells show similar phenotype with osteoblasts, periodontal ligament cells are thought to play an important role in alveolar bone remodeling. According to recent studies, receptor activation of nuclear factor $^{\kappa}B$ ligand (RANKL) and osteoprotegerin (OPG) are expressed in periodontal ligament cells during tooth movement. Also periodontal ligament cells is known to play an important role in the progression of periodontal disease. This study was designed how the expression of RANKL and OPG in periodontal ligament cells was regulated by IL-1 ${\beta}in$ the concentration of $0.01{\sim}10$ ng/ml. The results are as follows; 1. Periodontal ligament cells which stimulated by 1L-1 ${\beta}$ increased soluble RANKL synthesis by dose-dependent pattern in the concentration of $0.01{\sim}10$ ng/ml. 2. 1L-1 ${\beta}$ induced mRANKL expression in dose-dependent manner in the concentration of $0.01{\sim}5$ ng/ml. 3. mOPG expression was not to be influenced by 1L-1 ${\beta}$. These results suggested that rat periodontal ligament cells could regulate osteoclastogenesis by stimulation of production of RANKL.

• Journal of The Korean Society of Integrative Medicine
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• v.8 no.3
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• pp.53-62
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• 2020

Purpose :Periodontal ligament stem cells maintain tissue homeostasis in periodontal ligament. The purpose of this study was to determine the characteristics of periodontal ligament stem cells isolated from premolar teeth and observe protective effects against oxidative damage caused by Triethylene glycol dimethacrylate (TEGDMA) following treatment with N-acetylsysteine amide (NACA) drug known as enzymatic antioxidants. Methods : Primary periodontal ligament stem cell (PDSC) culture was performed from simply extracted human premolar of orthodontic patients. The characteristics of the primary cultured PDSCs was analyzed using the FACS system. PDSCs was incubated with TEGDMA and NACA. The cell proliferation and survival was determined using WST-1 assay. Collected data were analyzed using SPSS Window 20. Results : Primary cultured PDSCs grow on the floor and develop rapidly in a cluster form from up to 14 days. The morphology of PDSCs showed the spindle-shaped cells and grew directionally. FACS analysis, In addition, positive expression of visible cells were observed in mesenchymal stem cell biomarkers. PDLSCs cell viability was significantly decreased at high concentration in both 3 and 6 hours after TEGDMA treatment. We observed a decrease in the number of cells as well as a morphological change of PDLSCs. Antioxidative effect was notable since the death of PDLSC death was significantly inhibited compared to the control group at 24 and 48 hours after NACA treatment. Conclusion : Therefore, based on the results of this study, further research should be encouraged considering the development of clinical treatment methods using various antioxidants as well as regenerative engineering techniques utilizing periodontal ligament stem cells.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.33 no.5
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• pp.455-463
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• 2007

The present study was designed to evaluate effects of the commonly used NSAIDs(acetaminophen, aspirin, and ibuprofen) on human periodontal ligament cells. Human periodontal ligament cells were grown from a cell line provided by Kyungpook National University. Effects of NSAIDs on the proliferation of human periodontal ligament cells were assessed using MTT assays. And then $PGE_2$ concentrations were determined by ELISA and the changes of cellular configuration were found by electron micrograph. The results were as follows; 1. The MTT assay demonstrated that the commonly used NSAIDs(acetaminophen, aspirin, and ibuprofen) had not significant cytotoxic effect on human periodontal ligament cells. 2. NSAIDs inhibited the $PGE_2$ synthesis of human periodontal ligament cells compared with the control group. These inhibitory effects had no significant differences with NSAID type and concentration. 3. Electron micrographs of human periodontal ligament cells treated with NSAIDs showed more narrow and irregular shape.

• Journal of Periodontal and Implant Science
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• v.25 no.2
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• pp.239-251
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• 1995

The selective migration, attachment and proliferation of periodontal ligament cells are the desired goal of periodontal regeneration therapy. Fibronectin is well known for an attachment protein for dentin surface. Also, Fibroblast growth factor (FGF) is well known to enhance the periodontal regeneration. The purpose of this study was to evaluation the effect of fibronection and FGF on the attachment rate and the cellular activity. Human gingival fibroblast and periodontal ligament cells were cultured from the teeth extracted for non-periodontal reson. Cultured human gingival fibroblast and periodontal ligament cells in vitro were treated with fibronectin and FGF a various dosage and culture times. Cellular activity was examined by MTT assay. The results of this study was demonstrated that cell attachment rate of experimental group was under the control value at 1st, 2nd, 3rd incubation day. But, at 3rd incubation day, attchment value tended to return to the control value. In case of fibronectin alone application, cellular activity was decreased than that of control at 1st, 2nd incubation day. But 3rd day, cellular activity was returned to the control value. The activity of gingival fibroblast in FGF alone application was decreased thatn that of control at each incubation day. But activity of periodontal cell group was increased cell activities at 2nd, 3rd day. Additionally cellular activity of fibronectin & FGF combined application on gingival fibroblast group was similar to control value at incubation day. But activity of periodontal ligament cell group was increased at 2nd, 3rd day compared with control group.This study demonstrated that combined application of fibronectin & FGF induced the selective chemotaxis for periodontal ligament cell in vitro.

• Journal of Periodontal and Implant Science
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• v.26 no.1
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• pp.188-201
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• 1996

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.

• Journal of Periodontal and Implant Science
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• v.27 no.2
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• pp.291-303
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• 1997

Periodontal ligament cells may have a role in the regulation of hard and soft periodontal tissues, but their specific function has not yet to be determined. To evaluate further their role in periodontal regeneration, they were examined for osteoblast-like behavior. Periodontal ligament cells and gingival fibroblasts were primarily cultured from extracted premolar with non-periodontal diseases. Cells were cultured with DMEM at $37^{\circ}C$, 5% $CO_2$, 100% humidity incubator, and as a measure of cell characterization, it was examined that the morphology, alkaline phosphatase activity, collagen synthesis, and immunocytochemistry for osteonectin, osteocalcin, and collagen type I. Healthy periodontal ligament cells has more osteoblastic-like cell property in alkaline phosphatase activity. and collagen synthesis than gingival fibroblast. Immunocytochemistry localization explained that calcitonin were expressed in periodontal ligament cells only, and osteonectin and type I collagen were produced in both cells simultaneously. This results indicate that the growth characteristics of periodontal ligament cells and gingival fibroblasts exhibit some differences in proliferative rates and biochemical synthesis. The differences may help to calrify the role such cells play in the regenearation of periodontal tissues.

• Journal of Periodontal and Implant Science
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• v.37 no.1
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• pp.53-64
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• 2007

The aim of this study was to examine the possibility of periodontal ligament regeneration when autotransplantation was used by the periodontal ligament fibroblasts cultured on the acellular dermal matrix in teeth without a periodontal ligament. One minipig was used in this study. The mandibular and maxillary permanent incisors were ex-tracted for the culture of the periodontal ligament cells. The roots of the unextracted teeth were classified into a positive control group, in which the normal periodontal ligament was preserved. The roots of the extracted teeth were divided into the following two groups: The negative control group, in which the periodontal ligament had been removed and the acellular dermal matrix was not applied; and an experimental group, in which the periodontal ligament had been removed and periodontal ligament fibroblast cultured on an acellular dermal matrix was applied. The prepared teeth were transplanted, and completely submerged using physical barrier membranes. The animal was sacrificed 4 weeks after the autotransplant. The transplanted teeth were examined histologically. In this study, the periodontal ligament was normal in the positive control group, and ankylosis was discovered on the denuded root surface in the negative control group. Periodontal ligament-like connective tissue was found adjacent to the denuded root and the new cementum-like layer of hard tissue was formed in the experimental group. These results suggest that the periodontal ligament fibroblasts cultured on the acellular dermal matrix may play a role in regenerating the periodontal ligament-like tissue with new cememtum-like tissue formation.