• Journal of Periodontal and Implant Science
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• 제25권1호
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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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• 제26권2호
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• pp.414-428
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• 1996

The use of basic fibroblast growth factor which function as potent biologic mediators regulating numerous activities of wound healing has been suggested for the promotion of periodontal regeneration. The mitogenic effects of basic fibroblast growth factor on human periodontal ligament cells and human gingival fibroblasts were evaluated by determining the incorporation of 5-Bromo-2'deoxy-uridine into DNA of the cells in a dose -dependent manner. The cells which were prepared were the primary cultured gingival fibroblasts and periodontal ligament cells from human the fourth or sixth subpassages were used in the experiments. The cells which were seeded DMEM contain 10% FBS. The added concentrations of basic fibroblast growth factor were 0.1, 1, 10, 50, $l00{\eta}g/ml$ and basic fibroblast growth factor were added to the quiescent cells for 24 hours, 48 hours and 72 hours. They were labeled with $10{\mu}l/200{\mu}l$ 5Bromo-2'-deoxy-uridine for the last 6 hours of each culture. The results of the five determinants were presented as mean and S.D.. The results were as follows. : The DNA synthetic activity of human gingival fibroblasts was increased dose dependently by basic fibroblast growth factor at 24 hours, 48 hours and 72 hours. The similar mitogenic effects were at the 24 and 48 hours of basic fibroblast growth factor, but the DNA synthetic activity of human gingival fibroblasts generally decreased at 72 hours. The DNA synthetic activity of human periodontal ligament cells was increased dose dependently to $50{\eta}g/ml$ by basic fibroblast growth factor at 24, 48 and 72 hours, but the DNA synthetic activity decreased at $l00{\eta}g/ml$ of each hour. Generally the maximum mitogenic effects were at the 48 hours application of basic fibroblast growth factor. The DNA synthetic activity of human periodontal ligament cells generally decreased lower at 72 hours than at 24, 48 hours the application of basic fibroblast growth factor. In the comparison of DNA synthetic activity between human gingival fibroblasts and human periodontal ligament cells, human periodontal ligament cells had slightly higher proliferation activity than human gingival fibroblasts for a longer time at the high dosage of the basic fibroblast growth factor.In conclusion, basic fibroblast growth factor have important roles in the stimulation of DNA synthesis in human periodontal ligament cells and human gingival fibroblasts, and thus may be useful for clinical applications in periodontal regenerative procedures.

• Journal of Periodontal and Implant Science
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• 제25권2호
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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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• 제29권1호
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• pp.31-40
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• 1999

Many factors may affect periodontal changes during the physiologic conditions of woman(e.g. puberty, menstrual cycle, pregnancy, menopause). Recently many research has focused on the immunological changes of host, but the exact mechanism is not clear. Collagen is a major constituent of periodontium, and collagenase specifically digests the collagen and plays a role in destruction of periodontal tissue. So, I suppose that it participates with the cytokines in the inflammation of gingiva and vascular response during the changes of female sex hormones. Because there are some evidences of the existence of the receptors of estrogen and progesterone in the gingiva, it may be a target tissue of female sex hormones. In this experiment, gingival fibroblast and periodontal ligament cell were cultured in the presence of various concentrations of estrogen or progesterone corresponding to the menstrual cycle and pregnancy. Collagenase activity of the supernatant of culture media was determined by Spectrophotometric collagenase assay. The enzyme activity was calculated by the % decrease of the coated collagen. 1. The estrogen at both concentrations had no effect on the activity of collagenase of the gingival fibroblast. 2. The progesterone had some effect on the collagenase activity of the gingival fibroblast at low and high concentration of menstrual cycle, and elevated the enzyme activity at all range of pregnancy concentrations. 3. In periodontal ligament cells, estrogen elevated the enzyme activity at the early pregnancy concentration and progesterone elevated at the concentration just before menstruation. In this experiment, pregesterone elevated the collagenase activity of gingival fibroblast and periodontal ligament cells. But the mechanism of the up-regulation of the enzyme activity was not confirmed. The more experiments of direct effect of progesterone on gingival at the molecular level(e.g. northern blot analysis) can reveal the exact mechanism.

• Journal of Periodontal and Implant Science
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• 제37권1호
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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.

• Journal of Periodontal and Implant Science
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• 제26권1호
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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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• 제27권2호
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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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• 제28권1호
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• pp.17-35
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• 1998

Chitosan, with a chemical structure similar to hyaluronic acid, has been implicated as a wound healing agent. The purpose of this research was to evaluate the effects of chitosan on the characteristics of periodontal ligament cells, calvaria cells and gingival fibroblasts and to define the effects of chitosan on bone formation in vitro. In control group, the cells were cultured alone with Dulbecco's Modified Eagle's Medium contained with 10% Fetal bovine serum, 100unit/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. In experimental group, chitosan($40{\mu}g/ml$) is added into the above culture condition. And then each group was characterized by examining the cell proliferation at 1,3,5,7,9,12,15 day, the amount of total protein synthesis, alkaline phosphatase activity at 3, 7 day and the ability to produce mineralized nodules of rat calvaria cell at 11 day. The results were as follows : 1. At early time both periodontal ligament cells and calvaria cells in chitosan-treated group proliferated more rapidly than in non-treated control group, but chitosan-treated group of periodontal ligament cells at 9 days and calvaria cells at 12days showed lower growth rate than control group. Gingival fibroblast in chitosan-treated group had lower growth rate than in control group but the difference was not statistically significant (P< 0.01).2. Both periodontal ligament cells and calvaria cells in chitosan-treated group showed much protein synthesis than in control group at 3 days, but showed fewer than in control group at 7 days. Amount of total protein synthesis of gingival fibroblast didn't have statistically significant difference among the two groups(P< 0.01). 3. At 3 and 7 days, alkaline phosphatase activity of periodontal ligament cells and calvaria cells was increased in chitosan-treated group, but at 7 days there was not statistically significant difference among the two groups of calvaria cells (P< 0.01). Alkaline phosphatase activity of gingival fibroblast didn't have statistically significant difference among the two groups(P<0.01). 4. Mineralized nodules in chitosan-treated group of rat calvaria cells were more than in control group. In summery, chitosan had an effect on the proliferation, protein systhesis, alkaline phosphatase activity of periodontal ligament cells and calvaria cells, and facilitated the formation of bone. It is thought that these effects can be used clinically in periodontal regeneration therapy.

• International Journal of Oral Biology
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• 제30권3호
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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.

• International Journal of Oral Biology
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• 제32권2호
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• pp.67-73
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• 2007

Periodontitis is a chronic infectious disease that leads to periodontal destruction, and is one of the major causes of tooth loss in humans. The osteoclast differentiation factor (ODF), which is also known as the receptor activator of the NF-kB ligand (RANKL), is a surface-associated ligand on bone marrow stromal cells and osteoblasts. RANKL activates its cognate receptor, RANK, on osteoclast progenitor cells, which leads to the differentiation of mononucleated precursor cells. Osteoprotegerin (OPG) is a decoy receptor that is released from stromal cells and osteoblasts to inhibit the interaction between RANKL and RANK. Although the precise mechanism of bone loss in periodontitis is unknown, the differentiation and activation of osteoclasts by OPG-ODF-RANK signaling might play the role in periodontal bone destruction. The relationship between the concentration of sex hormones and the expression of ODF and OPG was examined by treating human gingival fibroblasts and periodontal ligament cells with the normal serum concentration of estrogen or progesterone during menstruation or at menopause. The ODF/OPG relative ratio was elevated at the concentration observed during ovulation in human gingival fibroblasts and at the concentration observed between ovulation and menstruation in periodontal ligament cells treated with estrogen. However, the ratio was <1 at all concentrations in both cells treated with progesterone. In the case of menopause simulated by estrogen depletion, the ratio was <1 in human gingival fibroblasts but >1 in periodontal ligament cells.