• 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.

• International Journal of Oral Biology
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• v.30 no.3
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• pp.105-110
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• 2005

Periodontitis is a chronic infectious disease that leads to the destruction, one of the major cause of tooth loss in human. Osteoclast Differentiation Factor(ODF), also called as Receptor activator of NF-${\kappa}B$ ligand(RANKL), a surface-associated ligand on bone marrow stromal cells and osteoblasts, activates its cognate receptor RANK on osteoclast progenitor cells, which leads to differentiation of these mononucleated precursor cells. Osteoprotegerin(OPG), a decoy receptor, is released from stromal cells and osteoblasts to inhibit the interaction between RANKL and RANK. The experiment for the effect of pregnancy on gingival health showed greater gingival inflammation and edema during pregnancy, despite similar plaque index. There should be many factors affecting the periodontal health in pregnancy. In this experiment, we examined the direct effects of sex hormones(estrogen and progesterone) on the ODF/OPG expression in human gingival fibroblasts and periodontal ligament cells at the serum concentration of pregnancy. The ratio was high in the 1st trimester of pregnancy by estrogen and in the late 2nd trimester by progesterone. Therefore, the local periodontal destruction might be accelerated by these hormonal effect on the periodontal cells.

• 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.40 no.6
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• pp.265-270
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• 2010

Purpose: The aim of this study was to investigate the immunomodulatory effects of canine periodontal ligament stem cells on allogenic and xenogenic immune cells in vitro. Methods: Mixed cell cultures consisting of canine stem cells (periodontal ligament stem cells and bone marrow stem cells) and allogenic canine/xenogenic human peripheral blood mononuclear cells (PBMCs) were established following the addition of phytohemagglutinin. The proliferation of PBMCs was evaluated using the MTS assay. The cell division of PBMCs was analyzed using the CFSE assay. The apoptosis of PBMCs was assessed using the trypan blue uptake method. Results: Periodontal ligament stem cells and bone marrow stem cells inhibited the proliferation of allogenic and xenogenic PBMCs. Both periodontal ligament stem cells and bone marrow stem cells suppressed the cell division of PBMCs despite the existence of a mitogen. No significant differences in the percentages of apoptotic PBMCs were found among the groups. Conclusions: Canine periodontal ligament stem cells have an immunomodulatory effect on allogenic and xenogenic PBMCs. This effect is not a product of apoptosis of PBMCs but is caused by the inhibition of cell division of PBMCs.

• The korean journal of orthodontics
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• v.42 no.5
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• pp.249-254
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• 2012

Objective: To investigate the stem cell-like characteristics of human periodontal ligament (PDL) stromal cells outgrown from orthodontically extracted premolars and to evaluate the potential for myogenic differentiation. Methods: PDL stromal cells were obtained from extracted premolars by using the outgrowth method. Cell morphological features, self-replication capability, and the presence of cell-surface markers, along with osteogenic, adipogenic, and chondrogenic differentiation, were confirmed. In addition, myogenic differentiation was induced by the use of 5-aza-2'-deoxycytidine (5-Aza) for DNA demethylation. Results: PDL stromal cells showed growth patterns and morphological features similar to those of fibroblasts. In contrast, the proliferation rates of premolar PDL stromal cells were similar to those of bone marrow and adipogenic stem cells. PDL stromal cells expressed surface markers of human mesenchymal stem cells (i.e., CD90 and CD105), but not those of hematopoietic stem cells (i.e., CD31 and CD34). PDL stromal cells were differentiated into osteogenic, adipogenic, and chondrogenic lineages. Myotube structures were induced in PDL stromal cells after 5-Aza pretreatment, but not in the absence of 5-Aza pretreatment. Conclusions: PDL stromal cells isolated from extracted premolars can potentially be a good source of postnatal stem cells for oromaxillofacial regeneration in bone and muscle.

• Journal of Periodontal and Implant Science
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• v.29 no.1
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• pp.15-30
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• 1999

The purpose of this study was to evaluate the effect of mixed extracts of aralia cortex and phellodendron cortex (P55A) on activities of human gingival fibroblasts and periodontal ligament cells in vitro. First experiment was done to evaluate the effect of P55A in normal condition. In control group, the cells($4.5{\times}10^4$ cells/ml) were cultured with Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum. In experimental groups, P55A was added to the above culture condition at the final concentrations of 0.1 ${\mu}g/ml$(Test group 1), 1 ${\mu}g/ml$(Test group 2) and 10 ${\mu}g/ml$(Test group 3). Then each group was tested for the cell proliferation rate at $\frac{1}{2}$, 2, 5 days, protein levels at 2, 5 days, and alkaline phosphatase activity at 2, 5 days. Second experiment was done to evaluate the effect of P55A in high glucose condition. 200 mg/dl glucose was added to the same culture condition of all groups in first experiment. Then each group was tested for the cell proliferation rate at $\frac{1}{2}$ , 2, 5 days, protein levels at 2, 5 days, and alkaline phoaphatase activity at 2, 5 days. The results were as follows ; 1. First experiment 1) As P55A concentration increased, cell proliferation rate increased significantly in test group 2 at 2 days, and test group 2 and 3 at 5 days in human gingival fibroblasts and periodontal ligament cells(P<0.05). 2) In human gingival fibroblasts, all test groups showed significantly increased protein levels as compared to control group at 5 days. In periodontal ligament cells, test group 2 and 3 showed significantly increased protein levels as compared to control group at 2, 5 days(P<0.05). 3) Alkaline phosphatase activity of human periodontal ligament cells increased as P55A concentration increased. The test group 2 and 3 showed significant increase as compared to control group at 5 days(P<0.05). 2. Second experiment 1) As P55A concentration increased, cell proliferation rate increased significantly in test group 2 at 2 days, and test group 2 and 3 at 5 days in human gingival fibroblasts and periodontal ligament cells(P<0.05). 2) In human gingival fibroblasts, test group 3 showed significantly increased protein levels as compared to control group at 2 days, and all test groups at 5 days. In periodontal ligament cells, test group 2 and 3 showed significantly increased protein levels as compared to control group at 2, 5 days(P<0.05). 3) Alkaline phosphatase activity of human periodontal ligament cells increased as P55A concentration increased. The test group 2 and 3 showed significant increase as compared to control group at 2 days, and all test groups at 5 days(P<0.05). From the above results, mixed extracts of aralia cortex and phellodendron cortex appeared to enhance cellular activities including cell proliferation rate, protein levels and alkaline phosphatase activity of human gingival fibroblasts and periodontal ligament cells in normal and high glucose condition. This study suggests that mixed extracts of aralia cortex and phellodendron cortex seem to be able to subside the inflammation of periodontal tissue and regenerate the destructed periodontal tissue.

• International Journal of Oral Biology
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• v.33 no.4
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• pp.131-135
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• 2008

Substance P (SP) is known to be expressed in the nerve fibers of dental pulp and periodontal tissues. It was recently reported that SP expression increased in response to orthodontic force. In the present study, we investigated the effect of SP on expression of mineralization markers and heme oxygenase-1 (HO-1) in human immortalized periodontal ligament (IPDL) cells. Cell viability was measured using a 3,4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT) assay. The expression of mineralization markers, including alkaline phosphatase (ALP), osteonectin (ON) and bone sialoprotein (BSP), and heme oxygenase-1 (HO-1) was assessed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis. SP did not significantly change human IPDL cell viability, with the exception of the 24 hour treatment group. Treatment of human IPDL cells with $10^{-10}$ to $10^{-4}M$ SP upregulated mineralization marker and HO-1 expression in a time- and concentration-dependent manner. Our results suggest that SP may modulate osteoblastic cell differentiation of human IPDL cells through a mechanism involving HO-1 expression.

• Journal of Periodontal and Implant Science
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• v.27 no.3
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• pp.459-468
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• 1997

Healing of periodontal tissues require the migration and proliferation of gingival fibroblasts and periodontal ligament cells. There is many evidences that the some agents like cytokines and polypeptide growth factors are mediate these cellular events in wound healing. Recently someone is interested in herbal drugs on periodontal tissue healing processes. The purpose of this study was to examine the effects of 4 herbal drugs, Carthami Flis, Moutan Redias Cortex, Scirpi Rhisoma, Seed of Carthamus tinctorius L. on human gingival fibroblasts and periodontal ligament cells. Periodontal ligament cells and gingival fibroblasts were primarily cultured from extracted premolar with non-periodontal diseases. The powder from extracted. herbal drugs were prepared with distilled water. Cells were cultured with DMEM at $37^{\circ}C$, 5% $CO_2$, 100% humidity incubator, and treated with each herbal drugs with proper concentration for 1, 2, and 3 days. The cell activity was determined by ELISA reader using MTT assay. There was the most significant elevation in $10^{-3}g/ml$ of almost herbal drugs on cellular activities. The result of this study demonstrated that Carthami Flis, Moutan Radicis Cortex, Scirpi Rhisoma, Seed of Carthamus tinctorius L. appears to have beneficial effect on healing process after periodontal treatment.

• Molecules and Cells
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• v.40 no.8
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• pp.550-557
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• 2017

The periodontal ligament (PDL) is the connective tissue between tooth root and alveolar bone containing mesenchymal stem cells (MSC). It has been suggested that human periodontal ligament stem cells (hPDLSCs) differentiate into osteo/cementoblast and ligament progenitor cells. The periodontitis is a representative oral disease where the PDL tissue is collapsed, and regeneration of this tissue is important in periodontitis therapy. Fibroblast growth factor-2 (FGF-2) stimulates proliferation and differentiation of fibroblastic MSCs into various cell lineages. We evaluated the dose efficacy of FGF-2 for cytodifferentiation of hPDLSCs into ligament progenitor. The fibrous morphology was highly stimulated even at low FGF-2 concentrations, and the expression of teno/ligamentogenic markers, scleraxis and tenomodulin in hPDLSCs increased in a dose dependent manner of FGF-2. In contrast, expression of the osteo/cementogenic markers decreased, suggesting that FGF-2 might induce and maintain the ligamentogenic potential of hPDLSCs. Although the stimulation of tenocytic maturation by $TGF-{\beta}1$ was diminished by FGF-2, the inhibition of the expression of early ligamentogenic marker by $TGF-{\beta}1$ was redeemed by FGF-2 treatment. The stimulating effect of BMPs on osteo/cementogenesis was apparently suppressed by FGF-2. These results indicate that FGF-2 predominantly differentiates the hPDLSCs into teno/ligamentogenesis, and has an antagonistic effect on the hard tissue differentiation induced by BMP-2 and BMP-4.

• The Journal of Korean Medicine
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• v.24 no.3
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• pp.35-44
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• 2003

Objective : This study was performed to evaluate the effects of Palmijihwang-hwan (Baweidehuang-wan) and Obaeja (Galla Rhois) on the regeneration of periodontal tissue. Methods : In this study, we used MC3T3-El cells, such as osteoblast-like cell lines and human periodontal ligament fibroblasts, for experimental material. We separated each type of cells into a control group and an experimental group. In the control group, the cells were cultivated for 48 hours with distilled water and media which contained 10% fetal bovine serum (FBS) and penicillin (l00unit/ml)-streptomycin ($l00{\mu\textrm{g}}/ml$) at $37^{\circ}$ in 5% $CO_2$ gas. In the experimental group, the cells were cultivated for 48 hours with Palmijihwang-hwan extract and Obaeja extract (concentrations $1{\mu\textrm{g}}/ml,{\;}25{\mu\textrm{g}}/ml,{\;}50{\mu\textrm{g}}/ml$) under the same conditions as the control group. Investigating the regeneration of periodontal tissue was performed by evaluating proliferation, the activity of alkaline phosphatase and the synthetic ability of proteins using those cultivated cells by means of microculture tetrazolium (MTT) assay, alkaline phosphatase substrate kit and protein assay kit. Results : 1. In vitro, Palmijihwang-hwan extract increased the proliferation of MC3T3-El cells. 2. In vitro, Obaeja extract increased the activity of alkaline phosphatase and the synthetic ability of protein in MC3T3-El cells and human periodontal ligament fibroblasts depending on Obaeja extract's concentration. Conclusion : Obaeja extract can be developed as a subsidiary medicine for the regeneration of periodontal tissue. Further studies to evaluate the different concentrations the Obaeja extract and clinical trials in vivo are suggested.