• Journal of Periodontal and Implant Science
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• 제29권4호
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• pp.821-838
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• 1999

On the basis of the evidences that electrical stimulation could enhance proliferation and differentiation of bone cells and promote healing and regeneration of bone, this study was performed to investigate the effects of electrical stimulation on human periodontal ligament cells and gingival fibroblasts in vitro, which also have important roles in regeneration of periodontium, and to evaluate the potential of clinical application of electrical stimulation. Human periodontal ligament cells and gingival fibroblasts were primarily cultured from the root surface of extracted premolar and the adjacent gingiva without periodontal diseases. In control group, the cells ($5{\times}10^4$ cells/ml)were incubated only in Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum. In test groups, electrical stimulation was given at the current intensity of $0.25{\mu]A$(test group 1), $1.0{\mu}A$(test group 2), and $2.5{\mu}A$(test group 3) for 12 hours to the same culture media with the control group. After 12 hour exposure of electrical stimulation, the cells were incubated for 2 and a half days(60 hours), and then each group of cells was analyzed for cell proliferation, protein level, and activity of alkaline phosphatase. The results were as follows ; 1. The Rate of cell proliferation of every test group increased significantly in both periodontal ligament cells and gingival fibroblasts, and in periodontal ligament cells, test group 3 showed significantly increased proliferation compared to the other test groups(p<0.05). 2. In the protein levels, neither periodontal ligament cell nor gingival fibroblast showed statistically significant differences between control and test groups. 3. The activity of alkaline phosphatase in periodontal ligament cells increased significantly in all test groups(p<0.05), but there were no significant differences between 3 test groups. In gingival fibroblasts, the activity of alkaline phosphatase increased significantly only in test group 3(p<0.05). From the above results, it is concluded that electrical stimulation may have beneficial effects on the regeneration of destructed periodontal tissue in regard of the stimulation of periodontal ligament cells and gingival fibroblasts as well as electrically stimulated bone formation that has been known, and that electrical stimulation may have the potential of clinical application.

• 대한치주과학회:학술대회논문집
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• 대한치주과학회 2000년도 제40회 종합학술대회 연제초록
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• pp.66-66
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• 2000

• Journal of Oral Medicine and Pain
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• 제19권2호
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• pp.57-71
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• 1994

Gingival fibroblasts were cultured and subjected to the test of Northern blot analysis for the demonstration of various mRNA expression in response to the low level laser treatment. For duplication of in vivo. Wound healing process, fibroblasts were pretreated with proinflammatory cytokine interleukin-1$\beta$(IL-1$\beta$) or mitogenic substance phorbol 12-myristate 13-acetate(PMA) prior to laser irradiation. The results were as follows : 1. By the laser irradiation, the gene expression of collagen type I was markedly increased I n gingival fibroblasts, especially in the case of PMA pretreatment. The gene expression of collagen type IV, however, was not only affected by laser irradiation but also by chemical cell stimulation. 2. Oncogene v-myc expression was affected by both laser irradiation and IL-1$\beta$ or PMA stimulation, But v-fos gene expression was not detected in any case of this experimental system. 3. Heat shock gene(Hsp 70)was expressed constiutively, but slightly increased by laser irradiation. 4. mRNA of fibroblast growth factor(FGF) was induced by both laser irradiation and IL-1$\beta$ or PMA treatment.

• Journal of Periodontal and Implant Science
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• 제27권3호
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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.

• Journal of Periodontal and Implant Science
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• 제27권4호
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• pp.909-922
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• 1997

The purpose of this study was to evaluate the effect of high glucose on prostaglandin E2 production in human gingival fibroblasts and periodontal ligament cells in vitro. In control group, the cells($5{\times}10^4\;cells/ml$) were cultured with Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum, 45mg/dl glucose. In experimental groups, glucose was added to the above culture condition at the final glucose concentrations of 100mg/dl(Test group 1), 200mg/dl (Test group 2) and 400mg/dl (Test group 3). Then each group was tested for the cell proliferation rate, protein levels, and prostaglandin E2 production at $\frac{1}{2}$, 1, 2, 5 days. The results were as follows : 1. As glucose concentration increased, cell proliferation rate decreased significantly at 1, 2, 5 days in human gingival fibroblasts and periodontal ligament cells(P<0.01). 2. In human gingival fibroblasts, test group 2 and 3 showed significantly decreased protein levels as compared to control group at 5 days (P<0.01). 3. In human periodontal ligament cells, as glucose concentration increased, protein levels decreased significantly at 2 days and 5 days(P<0.01). 4. Prostaglandin $E_2$ production in human gingival fibroblasts and human periodontal ligament cells significantly increased as glucose concentration increased(P<0.01). results at 5 days showed obvious difference as compared to those at 2 days. From the above results, high glucose appeared to affect cellular activities including cell proliferation rate, protein levels and enhance prostaglandin $E_2$ production. It was assumed that prostaglandin E2 production by high glucose enhances inflammatory reaction and has a toxic effect on human gingival fibroblasts and human periodontal ligament cells. This study suggests that periodontal disease in diabetic patient is related to prostaglandin $E_2$ production.

• 대한치과보철학회지
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• 제45권3호
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• pp.382-388
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• 2007

Statement of problem. The role of calcium sulfate in stimulating the growth of gingival soft tissue has been reported in few studies. Such a unique property of calcium sulfate could serve as a trouble-solving broker in compensating for the lack of soft tissues in various oral surgeries. Purpose. The purpose of this study was to compare the proliferating activities of human gingival fibroblasts seeded on various bone graft barrier materials of calcium sulfate, collagen, and polytetrafluorethylene (PTFE). Material and methods. Two calcium sulfates ($CAPSET^{(R)}$. and $CalForma^{(R)}$, Lifecore Biomedical Inc., St. Paul, Minnesota, USA), a resorbable natural collagen ($Bio-Gide^{(R)}$, Geistlich Pharma Ag., Wolhusen, Switzerland), and a non-resorbable PTFE ($TefGen-FD^{(R)}$, Lifecore Biomedical Inc., St. Paul, Minnesota, USA) served as the human gingival fibroblasts' substrates and comprised the four experimental groups, whereas the untreated floors of culture plastics were used in the control group, in this study. Cells were trypsinized, seeded, and incubated for 48 h. The proliferating activities of fibroblasts were determined by XTT and SRB assay and absorbance (optical density, OD) was measured. One-way ANOVA was used to analyze the differences in the mean OD values between the groups of CAPSET, CalForma, Bio-Gide, TefGen, and the control (p<0.05). Results. From the XTT assay, the mean OD value of the control group, the highest, was significantly greater than that of any of the four experimental groups followed by CalForma, CAPSET, TefGen, and Bio-Gide. Further, the mean OD value of CalForma, was significantly greater compared to that of Bio-Gide. From the SRB assay, Calforma showed the highest mean OD value, which was significantly greater than that of any other groups, followed by the control, CAPSET, Bio-Gide, and TefGen. The mean OD values of both the control and CAPSET were significantly greater compared to that of TefGen (p<0.05). Conclusion. Assessment of the viability and proliferation of cultured fibroblasts seeded and incubated for 48 h on various barrier-material substrates using XTT and SRB assay showed that calcium sulfate $CalForma^{(R)}$ promotes the proliferating activity of human gingival fibroblasts.

• Journal of Periodontal and Implant Science
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• 제24권2호
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• pp.357-375
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• 1994

The purpose of this study was to investigate the differences of histochemical characteristics in inflammatory fibrous gingival hyperplasia (FGH), phenytoin-induced gingival hyperplasia(PIGH), idiopathic gingival hyperplasia(IDGH) and control groups (healthy and inflammatory gingiva) by immunohistochemical method with various antibodies and histomorphological analysis. In immunohistochemical finding, antibodies to inflammatory cells (T/B lymphocytes, macrophages, other monocytes), proliferating cell nuclear antigen(PCNA), epidermal growth factor(EGF), factor VIII, and type I collagen were used. 1. The inflammatory infiltrates in FGH were less than those in inflammatory gingiva. The composition of inflammatory cells of PIGH was similar with that of FGH. IDGH showed a similar histologic findings with healthy gingival tissue. 2. In FGH, the number of fibroblasts and newly-formed collagen fibers was increased. No significant increase of fibroblasts and the dense accumulation of thick collagen fibers were seen in PIGH. The increase of fibroblasts and the dense accumulation of thick collagen were seen in IDGH. 3. PCNA-positive cells were localized mainly in the area accumulated with inflammatory cells and blood vessels, significantly increased in all hyperplastic tissue groups, and distributed evenly in IDGH. 4. The distribution of EGF were not observed in healthy gingiva but detected locally in area with confluent blood vessels,without significant difference between the other tissue groups. This results suggest that inflammation plays a significant role in inducing hyperplastic change of gingival tissue. While in DIGH, drug itself as well as inflammation seems to attribute to hyperplastic change.

• Journal of Periodontal and Implant Science
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• 제24권1호
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• pp.98-108
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• 1994

Gingival hyperplasia is frequently associated with the long-term use of phenytoin for control of convulsive disorder. The purpose of this study was to investigate on the effects of lipopolysaccharides (LPS), ursolic acid and oleanolic acid to phenytoin-induced cell activity in human gingival fibroblast. Human gingival fibroblasts were cultured form the healthy gingiva of orthodontic patients. Gingival fibroblasts were trypsinized and transferred to the weels of microtest plates. Fibroblast were cultured in growth medium added $5{\mu}g/ml$ of phenytoin, $5{\mu}g/ml$ of LPS, $10^{-7}M$ of ursolic acid and oleanolic acid. The passage number of cultured fibroblasts were fifth and eight. Cell morphology was examined by inverted microscope and the cell activity was measured by proliferation assay. Ursolic acid significantly modulated cell morphology into globular shape at the concentrantion of $10^{-7}M$ in the presence of phenytoin and LPS, and the cell activity was significantl decreased by ursolic acid or oleanolic acid regardless of the presence of phenytoin and LPS. These results suggested that the increased phenytoin-induced cell activity might be modulated by ursolic acid regardless of the presence of phenytoin and LPS. These results suggested that the increased phenytoin-induced cell activity might be modulated by ursolic acid or oleanolic acid. Further study is needed to clarify their toxicological effects on cellular modulation and mRNA expression change.

• Journal of Periodontal and Implant Science
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• 제23권3호
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• pp.622-634
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• 1993

Gingiva is remarkly sensitive to certain drugs. Especially, long term use of phentoin, dihydropyrydine (including nifedipine), cyclosporin and other drugs can be lead to pathologic changes in gingival tissue, especially in terms of proliferation of epithelium and connective tissue. Recent study in terms of proliferation of epithelium and connective tissue. Recent study is focused on the inhibition of drug-induced gingival hyperplasia by using medicaments. The purpose of this study was to investigate on the pharmacological effects of nifedipine, retinoic acid and glycyrrhetini acid to the activity in human gingival fibroblast. Human gingival fibroblasts were cultured from the healthy gingiva of orthodontic patients. Gingival fibroblasts were trypsinized and cultured in growth medium added $5{\mu}g/ml$ of nifedipine, $10^{+7}M$ of retinoic acid and glycyrrhetinic acid. The passage number of cultured fibroblasts were between fifth and eighth. The cell morphology was examined by inverted microscope and the cell acitivity was measured by the MTT assay. Nifedipine at the concentration of $5{\mu}g/ml$ was revealed significantly effective to increase the cell activity and lipopolysaccharide was cofactor to increase cell activity in the presence of nifedipine. However, retinoic acid was significantly effective on the globular change of cell morphology and loss of cell process regardless of the presence of nifedipine and LPS. Cell activity was significantly decreased by the glycyrrhetinic acid at the concentration of $10^-M$ regardless of the presence of nifedipine and LPS. These results suggested that the increased cell activity by nifedipine might be modulated by retinoic acid and glycyrrhetinic acid. Further study is needed to clarify on their toxicological effects during cellular modulation and mRNA expression change.

• 대한치주과학회:학술대회논문집
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• 대한치주과학회 2003년도 제43회 종합학술대회 연제초록
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• pp.53-54
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• 2003