• Maxillofacial Plastic and Reconstructive Surgery
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• v.28 no.5
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• pp.445-453
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• 2006

Cyclosporine A (CsA) is a powerful immunosuppresive agent used to prevent graft rejection of organ and treat autoimmune disease. One of the major side effects associated with CsA treatment is the development of gingival overgrowth. The purpose of this study was to investigate the mRNA expression and association of the several growth factors in gingival overgrowth induced by CsA, respectively. Gingival fibroblasts were obtained from gingival tissues of healthy donor and the patients treated with CsA. The cultured gingival fibroblasts were incubated with increasing concentrations of CsA for 24 hours, and the expression of MMP-1, TIMP-1, $TGF-{\beta}_1$, p21 were determined by reverse transcription-polymerase chain reaction (RT-PCR). The expressions of MMP-1 was slightly increased according to the concentration of treated CsA, but there was no statistical significance. TIMP-1 showed the increased expression at the CsA concentration of 250 and 500 ng/ml and significantly decreased at the CsA concentration of 750ng/ml. $TGF-{\beta}_1$ showed the increased expression at the CsA concentration of 500 and 750 ng/ml. The expression of p21 was not changed significantly. We concluded that the gingival hyperplasia induced by CsA was more related with $TGF-{\beta}_1$ than MMP-1 or TIMP-1 on gingival collagen metabolism in patients treated with CsA.

• Journal of Periodontal and Implant Science
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• v.23 no.1
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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.

• Restorative Dentistry and Endodontics
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• v.17 no.2
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• pp.263-286
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• 1992

This study was performed to evaluate and compare the cytotoxic effects of five root canal sealers to several different cell lines. Five root canal sealers were AH-26, N2, Sealapex, Tubliseal, and Vitapex. Each sealers were mixed according to the manufacturer's instructions, and culture media were added to each sealers immediately after mixing (the immediate group) and after three days (the third day group) and seven days (the seventh day group) respectively. And every sealer solutions were diluted to 1:1, 1:2, 1:3 and 1:4. Three different permanent cell lines (HEp-2, McCoy, MRC-S) and human gingival fibroblasts and mononuclear cells were challenged by each sealer solution and the cytopathic effects were evaluated using MTT-ELISA, MTT-microscopy, and lactate dehydrogenase (LD) activity. The results were as follows: 1. In HEp-2 and MRC-5 cells, Vitapex was the least cytotoxic sealers. 2. AH-26 showed mild cytotoxic effects to HEp-2, gingival fibroblast and mononuclear cells. 3. N2 was the most toxic sealer to gingival fibroblast and it showed relatively strong cytotoxicity to HEp-2, McCoy and MRC-S cells. 4. Tubliseal showed strong cytotoxic effects to HEp-2, McCoy, MRC-S, and mononuclear cells. 5. Sealapex showed strong cytotoxic effect to HEp-2, McCoy, and gingival fibroblasts.

• Journal of Periodontal and Implant Science
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• v.26 no.3
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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 Korean Dental Science
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• v.5 no.1
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• pp.1-6
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• 2012

Patients with gingival overgrowth are easily seen in dental clinics. Cyclosporin-A (CsA), a widely prescribed immunosuppressant induces gingival overgrowth in up to 35% of patients with medical history of organ transplantation. The immunosuppressant CsA can transform genetic expression of gingival fibroblasts, resulting in gingival overgrowth. Meticulous plaque control is recommended for treatment of gingival overgrowth. Substitution of the drug or surgical procedures such as gingivectomy and periodontal flaps should be considered after re-evaluation. Azithromycin is often recommended as a supplementary drug to reduce this side effect. Recent studies show that tacrolimus can be a more economic, efficient and safe substitute for CsA.

• Journal of Periodontal and Implant Science
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• v.38 no.sup2
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• pp.395-404
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• 2008

Purpose: Osteoprotegerin (OPG) is a secreted glycoprotein and a member of the tumor necrosis factor (TNF) receptor family that inhibits bone resorption by suppressing osteoclastogenesis. Gingival fibroblasts (GF) play a role in periodontal disease progression, and the purpose of this experiment was to evaluate influence of osteotropic factors on the expression of osteoprotegerin mRNA in these cells. Materials and Methods: In this experiment, the influence of osteoclastogenic factors, interleukin-1 beta (IL-$1{\beta}$), TNF-$\alpha$, prostanglandin E2 ($PEG_2$). parathyroid hormone (PTH) and 1$\alpha$, 25-dihydroxyvitamin $D_3$ on the expression of osteoprotegerin mRNA in GF was studied by Northern blot hybridization. Results: As expected, $PEG_2$ tended to inhibit OPG levels and this was most prominent at 24 hours of culture with $10^{-7}M$ of $PEG_2$. TNF-$\alpha$ at 10ng/ml and also at 25ng/ml decreased OPG levels to almost 30% of the control at 24 hours. This contrasts with reports of increased OPG levels from osteoblast/stromal cells and gingival fibroblasts stimulated by TNF-$\alpha$. Decrease of OPG levels with $PEG_2$ and TNF-$\alpha$ suggests a pathway whereby these mediators exert their resorptive effects. However, OPG levels were increased almost 3-fold at 24 hours with IL-1$\beta$(1 to 15ng/ml) and increased 1.4 fold with 24-hour treatment of $10^{-7}M$ PTH. Conclusion: Increase of OPG levels suggests that these 'osteoclastogenic' factors act in more complex ways and may act to inhibit bone resorption in inflammatory periodontitis. This result supports the role of OPG as a negative feedback mechanism in osteoclastic activity.

• Journal of Periodontal and Implant Science
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• v.30 no.4
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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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• v.31 no.1
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• pp.109-122
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• 2001

Gingival fibroblasts(GF) and periodontal ligament fibroblasts(PDLF) are the major cellular components of periodontal soft connective tissues, but the precise molecular biological differences between these cells are not yet known. In the present study, we investigated the expression of S100A4, S100A2 calcium-binding protein and osteoblast-specific factor 2(OSF-2, Periostin) mRNA in GF and PDLF in vitro through the process of reverse transcription-polymerase chain reaction(RT-PCR) and Northern blot analysis in each. Human GF and PDLF were isolated from the gingival connective tissue and the middle third of freshly extracted healthy third molars. They were cultured in Dulbecco's Modified Eagle Medium(DMEM) containing 10% fetal bovine serum and cells in the third passage were used in the experiments. After extracting total RNA from cultured cells, RT-PCR and Northern analysis were performed using S100A4-, S100A2- and Periostin-specific oligonucleotide primers and subcloned cDNA probes in each. In PT-PCR and Northern analysis, the expression of S100A4 and Periostin mRNA in GF was slightly detectable. Interestingly, the expression of S100A4 and periostin mRNA in PDLF was much higher than that in GF. On the other hand, S100A2 mPNA was highly expressed in both GF and PDLF. Since there was a marked difference of S100A4 and Periostin expression between GF and PDLF in vitro, these data suggest that S100A4 and periostin could be used as a useful marker for distinguishing cultured gingival fibroblasts and periodontal ligament cells.

• Journal of Periodontal and Implant Science
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• v.30 no.4
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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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• v.30 no.1
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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.