• Journal of Periodontal and Implant Science
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• 제30권1호
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• pp.77-91
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• 2000

The purpose of this study was performed to evaluate the effect of Armeniacae Semen extracts on human gingival fibroblasts and periodontal ligament cells in vitro. A experiment was done to evaluate the effect of Armeniacae Semen extracts in high glucose media. $400mg/d{\ell}$ glucose was added to the culture media of all groups. In control group, the cells($4.5{\times}10^4cells/ml$) were cultured with Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum. In experimental groups, Armeniacae Semen extracts was added to the above culture media at the final concentrations of $1{\mu}g/m{\ell}$(Test group 1) and $l0{\mu}g/m{\ell}$(Test group 2). Then each group was tested for the rate of cell proliferation at 1, 2, 5 days, protein levels at 2, 5 days, and alkaline phosphatase activity at 2, 5 days. The results were as follows ; 1. Under the high glucose condition 1)As centration of Armeniacae Semen extracts increased, the rate of cell proliferation decreased significantly in test group 2 at 5 days in human gingival fibroblasts, but increased significantly in test group 2 at 5 days in human periodontal ligament cells(P<0.05). 2)In human gingival fibroblasts, test group 2 showed significantly decreased protein levels as compared to control group at 5 days. In periodontal ligament cells, test group 1 and 2 showed not 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 concentration of Armeniacae Semen extracts increased. The test group 1and 2 showed significant increase as compared to control group at 5 days(P<0.05). From the above results, Armeniacae Semen extracts appeared to enhance cellular activities including the rate of cell proliferation, protein levels and alkaline phosphatase activity of selectively human periodontal ligament cells in high glucose media. This study suggests that Armeniacae Semen extracts seem to be able to subside the inflammation of periodontal tissue and regenerate the destructed periodontal tissue.

• Journal of Periodontal and Implant Science
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• 제30권4호
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• pp.869-885
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• 2000

Fibroblasts are major cellular components of gingiva and periodontal ligament. They regulate the healing process after surgery or injury. Recently, many natural medicines, whose advantages are less side effects and possibility of long-term use, have been studied for their capacity, their anti-bacterial and anti-inflammatory effects and regenerative potential of periodontal tissues. Sophorae radix have been traditionally used as an anti-bacterial and antiinflammatory drug in oriental medicine. The purpose of present study was to investigate the effects of Sophorae radix extract on cell cycle progression and its molecular mechanism in human gingival fibroblasts. Sophorae radix extracts($100{\mu}g/ml$) notably increased cell proliferation and cell activity in the human gingival fibroblasts as compared to non-supplemented controls. There was an increase in the S phase and a decrease in the G1 phase in $100{\mu}g/ml$ of Sophorae radix extracts group as compared to non-supplemented controls. The level of cyclin E and cdk 2 protein in test group was higher than that of control groups. But that of cyclin D, cdk 4, and cdk 6 was not distinguished from controls. The level of p53 protein in test group was lower than that of controls, whereas that of p21 was not different. The level of pRB protein in test group was higher than that of controls, whereas that of p16 was lower. These results indicate that the increase of cell proliferation by Sophorae radix extracts may be due to the increased expression of cyclin E and cdk 2, and the decreased expression of p53 and p16 in human gingival fibroblasts.

• Journal of Periodontal and Implant Science
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• 제30권1호
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• pp.145-157
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• 2000

Cytotoxic substances in dental calculus and root cementum of periodontally diseased teeth inhibit new attachment and regeneration. The purpose of scaling and root planing is to remove pathologic structures harboring these cytotoxic substances in order to create a biologically acceptable root surface. However, these procedures inevitably leave a non-biocompatible smear layer. Conventionally, the smear layer has been removed with low pH etching agents such as citric acid, phosphoric acid and tetracycline hydrochloride(TC). Lately, a supersaturated neutral pH etching solution of ethylene diamine tetraacetic acid(EDTA) has been found to be as effective as low pH etchants with respect to smear removal and to be superior in exposing root surfaceassociated collagen. The aim of the present study was to determine the effect of root surface treatment using EDTA on the initial attachment of human gingival fibroblasts. 27 human teeth, extracted due to severe periodontitis, were cut into dentin slices after root planing. The specimens were divided into TC group(treated with $50㎎/m{\ell}$ tetracycline-HCl, pH 1.52), EDTA group(treated with 17% EDTA, pH 7.4), and non-treated control group. After sterilization, 5th subcultured human gingival fibroblasts were seeded in each culture well containing a prepared root slice and incubated for 15 min., 60 min., and 4 hours in 5% $CO_2$ incubator at $37^{\circ}C$. At each incubation time, the number of attached fibroblasts were counted on the microphotographs taken at a magnification of x100. The difference of the number of attached cells between groups was statistically analyzed by the ANOVA followed by Duncan test in SPSS/PC＋programs. The results were as follows : 1. After incubation for 15 min, the attached cells were significantly more in EDTA group and TC group than non-treated control group(p<0.05), but there was no significance in the difference between EDTA group and TC group(p>0.1). 2. After incubation for 60 min and 4 hours, there was no significant difference in the number of attached cells between all groups(p>0.1). 3. In both EDTA group and TC group, there was no significant difference in the number of attached cells between different incubation(p>0.1). But in control group, the number of attached cells was significantly increased after incubation for 60 min, compared with incubation for 15 min(p<0.05). The above results suggest that root surface treatment using EDTA could enhance the initial attachment of gingival fibroblasts to root surface as effective as tetracycline-HCl.

• Journal of Periodontal and Implant Science
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• 제29권1호
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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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• 제30권1호
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• pp.31-37
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• 2005

Treponema maltophilum, a Group IV oral spirochete, is associated with periodontitis and endodontic infections. In this study we analyzed the functional role of the major surface protein of this organism (MspA) in human gingival fibroblasts (HGFs). The full-length gene encoding MspA was cloned and expressed in Escherichia coli by using the expression vector pQE-30. The recombinant protein (rMspA) was purified by affinity chromatography with nickel-nitrilotriacetic acid agarose and possible contamination of E. coli endotoxin in rMspA was removed by using polymyxin B-agarose. rMspA significantly induced the expression of pro inflammatory cytokines like IL-6 and IL-8 and intercellular adhesion molecule (ICAM)-1 in HGFs, when analyzed by reverse transcription-PCR, flow cytometry, and enzyme-linked immunosorbent assay. Our results indicate that MspA of T. maltophilum may play an important role in amplifying the local immune response by upregulating the expression of proinflammatory cytokines and ICAM-1.

• 한국진공학회지
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• 제12권S1호
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• pp.10-14
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• 2003

Coatings of hydroxyapatite (HA) and tricalcium phosphate/HA (TCP/HA) on titanium were fabricated by ion beam assisted deposition (IBAD). Significant effect of the Ca-P coatings on human Gingival Fibroblasts (HGFs) attachment and formation of type I collagen were found by using immunofluorescence microscope. TCP/HA and HA coatings exerted more HGFs attachment and collagen I formation. Comparing with HA coating, TCP/HA coating exhibited better responses during the late period of the tests. This investigation indicated that this surface modification method may enhance the biological seal at the cervical level of the titanium dental implants.

• Journal of Periodontal and Implant Science
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• 제26권3호
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• pp.669-679
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• 1996

Gingival overgrowth is a well known side effect of several drugs, including nifedipine, phenytoin, cyclosporin, dilitiazem, verapamil. A number of studies have been performed to investigate the mechanism by which nifedipine(a calcium channel blocking agent) affects the gingival tissue. The aim of the present work was to investigate the effect of nifedipine on healthy gingival fibroblasts with special emphasis on determining the changes in cellular proliferation and protein and collagen synthesis. Gingival fibroblasts were obtained from the explants of healthy gingiva of extracted 3rd molars or premolar teeth extracted from the patients for orthodontic treatment. To evaluate the effect of nifedipine on cell proliferation, the cells were seeded at a cell density of $1{\times}10^4$cells/well in 24-well culture plates and treated with 100 and 200ng/ml of nifedipine for 10days. After trypsinization, the cells were counted with a haemocytometer on 1st, 3rd, 5th, 7th and 10th days. Then, MTT assay was carried out. For total protein and percent collagen synthesis, $3{\mu}Ci/ml$ $^3H-proline$ was added to each well for the final 4 hours of the incubation period. The results indicate that nifedipine does not influence cell proliferation in healthy gingival fibroblast in vitro and has a specific effect in reducing total protein and percent collagen synthesis. On the above the findings, exogenous nifedipine does not influence on healthy human gingival fibroblast proliferation and protein and collagen synthesis.

• Journal of Periodontal and Implant Science
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• 제30권4호
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• pp.895-913
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• 2000

It is becoming increasingly clear that human gingival fibroblasts(HGF) may play a role in regulating immune responsiveness in inflammatory periodontal lesions. Stimulation of HGF with locally-secreted T cell cytokine $IFN_{\gamma}$ induces human leukocyte antigen class II(HLA II) expression on HGF, which is one of the characteristic feature of professional antigen presenting cells(pAPC). However, $IFN_{\gamma}$-treated HGF and other nonprofessional antigen presenting cells(npAPC) are known to be ineffective or less effective antigen presenter to resting T cells. This study, therefore, was undertaken in an effort to elucidate the differences in expression of cell surface molecules between npAPC in periodontal tissues, such as HGF and periodontal ligament fibroblasts(PDLF), and pAPC such as monocytes/macrophages. Using flow cytometry, the levels of cell surface expression of HLA-D, ICAM-1, LFA-3, and B7-1, which are involved in antigen presentation, were determined in HGF, PDLF and human myelomonocytic cell line THP-1. $IFN_{\gamma}$ clearly induced HLA-D expression on both of fibroblasts and monocytes dose dependently. However, expression level on monocytes were 4 to 5 times higher than that on fibroblasts, and induction rate was faster in monocytes than in fibroblasts. The levels of ICAM-1 expression on fibroblasts and monocytes were enhanced by $IFN_{\gamma}$ in a dose dependent manner. On the other hand, the expression of LFA-3 molecule, which could be detected in fibroblasts and monocytes without cytokine stimulation, was no more enhanced by addition of $IFN_{\gamma}$. B7-1, important costimulatory molecule in T cell activation and proliferation, was not detected on both of fibroblasts and monocytes even when stimulated with $IFN_{\gamma}$, except on monocytes fully differentiated by pretreatment of PMA and treated by $IFN_{\gamma}$. These results suggest that delayed expression of HLA-D and absence of B7-1 on $IFN_{\gamma}$ - treated fibroblasts may at least in part be involved in the ineffectiveness of fibroblasts as primary APC. And it is postulated that although periodontal fibroblasts may not serve as primary APC in normal periodontium, sustained expression of HLA II on ubiquitous fibroblasts in inflammatory lesions may perpetuate immune responses and produce chronic inflammation and tissue injury.

• Journal of Periodontal and Implant Science
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• 제21권1호
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• pp.179.2-179.2
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• 1991

• Journal of Periodontal and Implant Science
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• 제23권1호
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• pp.135-143
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• 1993

The investigation was undertaken to determin the altered function and structure of granulation tissue fibroblasts. Human granulation-tissue fibroblasts were cultured from periodontal chronic inflammatory lesions (SBI index : above 3) and compared with healthy gingival connective tissues fibroblasts a control(SBI index : below 1). Granulation tissue fibroblasts proliferated with a slower growth rate and exhibited larger cell size than control cells. Total protein profile of granulation tissue fibroblasts was almost identical to that of control cells with some exception. These results support tha theory that granulation tissue fibroblasts represent a distinct phenotype of fibrotic cells.