• Journal of Periodontal and Implant Science
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• 제29권4호
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• pp.979-995
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• 1999

본 연구에서는 치주질환과 관련이 깊은 것으로 알려진 구강내 spirochetes 균중 Treponema denticola 분쇄액(TDC)과 가장 최근에 분리 배양된 Treponema lecithinolyticum 분쇄액(TLC)이 치은섬유아세포의 cytokine 분비 및 matrix metalloproteinase(MMP) 활성에 미치는 영향을 알아 보기 위하여 균의 분쇄액을 치은섬유아세포에 처리한 후 Interleukin-6(IL-6)와 $Interleukin-1{\beta}(IL-1{\beta})$의 분비 증가 여부를 ELISA를 통하여 측정하였으며, 또한 gelatinase zymography와 gelatin 분해능 측정을 통하여 교원질 분해 효소의 하나인 pro-MMP-2(progelatinase A)의 활성화 여부를 측정한 결과 다음과 같은 결론을 얻었다. 1. TDC와 TLC가 치은섬유아세포의 IL-6 분비에 미치는 영향을 살펴 본 결과, TDC 와 TLC 처치군에서 세균 분쇄액이 없는 비처치군에 비해 IL-6 분비량이 증가하였으며 유의성 있는 차이가 있었다(p<0.05). 2. TDC 와 TLC로 처리한 치은섬유아세포의 $IL-1{\beta}$ 분비는 측정 가능치(1pg/ml)이하의 분비량이 관찰되었다. 그러므로 $IL-1{\beta}$의 분비에는 영향이 없는 것으로 보인다. 3. 치은섬유아세포에서 분비되는 분자량 72 kDa의 pro-MMP-2가 TDC와 TLC에 의해 활성형으로 발현되어 zymography상에서 62kDa의 위치에 clear band로 나타났다. 4. 치은섬유아세포가 분비하는 MMP-2의 gelatin 분해능이, TDC와 TLC 처치군에서 비처치군보다 높게 나타났으며 유의 성 있는 차이가 있었다(p<0.05). 5. TDC 처치군에서는 gelatin 분해능에 있어서 세균 자체의 serin protease의 영향이 있었으나 TLC 처치군에서는 치은섬 유아세포의 MMP에 의해서만 gelatin이 분해되었다. 이상의 결과를 보아 TDC와 TLC는 치은섬유 아세포를 자극하여 IL-6 의 분비는 증가시킬 수 있으나 $IL-1{\beta}$의 분비에는 영향을 미칠 수 없으며, 치은섬유아세포에서 분비되는 pro-MMP-2를 활성형으로 발현시켜 결합조직의 파괴를 야기함으로서 치주 질환의 병인론에 기여할 수 있음을 확인하였다.

• Journal of Oral Medicine and Pain
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• 제12권1호
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• pp.17-26
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• 1987

In order to investigate the biostimulatory effect of low power density laser radiation in vitro, human gingival fibroblasts were cultured in MEM in which experiment groups respectively were made to 30 sec, 60 sec and 90 sec group. The experiments were performed by cell count, DNA and protein content measurements after experimental groups were irradiated with GaAlAs laser every day by forth day and then control group and experimental groups were compared. The results were as follows: 1. Cell counts of experimental groups were increased with exposure time, but showed no significance (P>0.05). 2. When the protein contents were compared, there was a very significant increase in 90 sec. experimental group (P<0.01). 3. When the DNA contents were compared, there was a significant difference only between control and 70 sec. group (P<0.05).

• Restorative Dentistry and Endodontics
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• 제17권2호
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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.

• 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.

• 대한의생명과학회지
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• 제10권3호
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• pp.269-273
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• 2004

This study was designed to investigate the effect of ferulic acid on cell viability and cell adhesion activity in normal human gingival fibroblasts. The cell viability and cell adhesion activity of ferulic acid was measured by MTT assay or XTT assay, respectively, after normal human gingival fibroblasts were treated with or without ferulic acid for 48 hours. The cell viability of ferolic acid on normal human gingival fibroblasts did not show any decreasement by MTT assay and also, cell adhesion activity did not decreased by XTT assay, respectively, compared with control after cells were treated with various concentrations of ferolic acid for 48 hours. MTT/sub 50/ and XTT/sub 50/ were 2,130.0 μM and 1,773.7 μM ferolic acid, respectively. These results suggest that ferolic acid is non-toxic to normal human gingival fibroblasts by showing no significant differences in the cell viability and the adhesion activity compared with control by colorimetric assay.

• Journal of Oral Medicine and Pain
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• 제18권2호
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• pp.97-106
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• 1993

This study was performed to investigate the effects of infrared and visible light laser irradiation on cell viability of human gingival fibroblast. For the present study, the author used cultured fibroblast originated from sound gingiva which were fifth of sixth passage. Laser machine utilized here were stomalaser which irradiate infrared (GaAs diode) and red (HeNe) laser in turn with pulse wave pattern or continuous wave pattern, and the machine had several frequency mode presented by regeneration, relaxation and analgesic modalities. Cultured fibroblast samples were divided by this modalities of cell counts and laser exposure time which were 7-seconds of 150 seconds, respectively. 1 day after laser irradiation, each cell-well was treated with MTT and measured optical density with ELISA. The obtained results were as follows : 1. There was a tendency of increasing optical density in proportion to irradiation time in groups of $1\times10^4$ cell per well but in groups of $5\times10^3$ cell per well, reverse phenomena were observed. 2. The difference of optical density according to frequency modalities were not showed significantly except several cases in groups of $5\times10^3$ cell per well. 3. In general, cell viability of cultured human gingival fibroblast wer not showed consistent feature by low level laser irradiation.

• 대한통합의학회지
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• 제7권4호
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• pp.61-69
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• 2019

Purpose : Human gingival fibroblast cell is one of the the main cell types in periodontal tissue, which they can show anti-inflammatory activity through the production of numerous lines of inflammatory mediators such as inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and interleukins. Porphyromonas gingivalis, one of the oral pathogens, has reported to play a critical role in the development of periodontal diseases. This study aimed to investigate anti-inflammatory and antioxidative activities of Gracilaria textorii ethanol extract (GTEE) in P. gingivalis derived lipopolysaccharide (LPS-PG) stimulated human gingival fibroblast (HGF)-1 cell line. Methods : In order to analyze anti-inflammatory and antioxidative activities of GTEE in HGF-1 cell line, NOS enzyme activity, expression levels of iNOS, COX-2, NAD(P)H quinone dehydrogenase (NQO)1 and their transcription factors were estimated by Griess reaction and western hybridization. Results : LPS-PG induced overexpression of iNOS and COX-2, which was significantly attenuated by GTEE treatment in a dose-dependent manner without any cytotoxicity. In addition, intracellular NOS activity was in accordance with the result of iNOS expression. Due to important role in the regulation of inflammatory responses, phosphorylated status of p65 and c-jun, each subunit of nuclear factor (NF)-κB and activator protein (AP)-1, was also dose-dependently ameliorated by GTEE treatment. One of phase II enzymes, NQO1, and its transcription factor, nuclear factor erythroid 2-related factor 2 (Nrf2), were analyzed since elevated phase II enzyme expression inhibited inflammatory response, which was significantly elevated by GTEE treatment in HGF-1 cell line. Conclusion : In conclusion, GTEE mitigated LPS-PG-stimulated inflammatory responses by attenuating NF-κB and AP-1 activation as well as accelerating NQO1 and Nrf2 expression in HGF-1 cell line. These results indicate that GTEE might be utilized a promising strategy for potential anti-inflammatory agent in periodontal diseases.

• International Journal of Oral Biology
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• 제45권2호
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• pp.64-69
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• 2020

Short-chain fatty acids (SCFAs) such as acetate, propionate, and butyrate are secondary metabolites produced by anaerobic fermentation of dietary fibers in the intestine. Intestinal SCFAs exert various beneficial effects on intestinal homeostasis, including energy metabolism, autophagy, cell proliferation, immune reaction, and inflammation, whereas contradictory roles of SCFAs in the oral cavity have been reported. Herein, we found that low and high concentrations of SCFAs induce differential regulation of intracellular Ca²⁺ mobilization and expression of pro-inflammatory cytokines, such as interleukin (IL)-6 and IL-8, respectively, in gingival fibroblast cells. Additionally, cell viability was found to be differentially regulated in response to low and high concentrations of SCFAs. These findings demonstrate that the physiological functions of SCFAs in various cellular responses are more likely dependent on their local concentration.

• Journal of Periodontal and Implant Science
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• 제29권4호
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• pp.803-823
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• 1999

In order to reveal immunopathogenesis of periodontal tissue destruction, it is important to clarify the molecular mechanism of trafficking and retention of activated leukocytes, including monocytes/macrophages. Gingival fibroblasts may be involved in the regulation of inflammatory cell accumulation in the extravascular periodontal connective tissues via cytokine production and surface expression of adhesion molecules. In this study, it was investigated the molecular basis for the adhesive interactions between monocytes and fibroblasts such as peri-odontal ligament fibroblast(PDLF), human gingival fibroblast(HGF), and human dermal fibroblast(HDF). First, it was examined the evidence whether monocyte-fibroblast cell contact may cause signal transduction in fibroblasts. Being directly in contact with fixed human monocyte cell line THP-1, or U937, upregulation of IL-6 production, $TNF-{\alpha}$ mRNA expression and increased cell proliferation could be seen for fibroblasts. IL-6 production induced by monocyte- fibroblast coculture were further increased when fibroblasts had been pretreated with $IFN-{\gamma}$ or $IL-1{\beta}$ , and monocytes with LPS. Next, it was examined the expression of ICAM-1 which has been known to be involved in accumulation and activation of leukocytes in inflammatory diseases such as periodontitis. ICAM-1 was upregulated up to 10-fold on PDLF, HGF, and HDF by exposure to $IFN-{\gamma}$ or $IL-1{\beta}$. Furthermore, anti-ICAM-1 monoclonal antibody clearly blocked cocultureinduced IL-6 production by fibroblasts, suggesting that $ICAM-1/{\beta}_2$integrin pathway is involved in periodontal fibroblastmonocyte interaction. Overall, these findings provide evidence that periodontal fibroblasts could be involved in the accumulation and retention of monocytes/macrophages in periodontal inflammatory lesion at least in part by ICAM-1 expression. In addition, periodontal fibroblast-monocyte interaction could cause activation signals in fibroblasts intracellularly which result in cytokine production and cell proliferation. Thus, periodontal fibroblasts are speculated to play an important role in immunoregulation and tissue destruction in chronic periodontal diseases by interaction with monocytes/macrophages.

• Journal of Oral Medicine and Pain
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• 제17권2호
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• pp.129-149
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• 1992

In order to verify the acceleration effect of low level laser (LLL) on oral mucosal wound healing process at cell biological level, the author studied growth pattern and ultrastructure in human gingival fibroblasts flashed by GaAlAs LLL-830 nm, 15mW for 10 minutes/flash one to three times at interval of 3-4 days through the evaluation of cell growth rate, protein conent/cell, DNA content/cell and ultrastructural changes for 14 days. The results were as follows : 1. The growth rate in gingival fibloblasts treated by LLL showed 4 orderly stages-decreasing stage after LLL treatment, acute increasing stage 3 days after LLL treatment, restring stage and recovering stage. 2. The effect of multiple flashes on LLL at interval of 3 days more or less was not proportional to times of flash on acceleration of growth in gingival fibroblasts. 3. The total protein content per gingival fibroblast was not significantly changed by LLL treatment in comparison with control group. But some kinds of protein which might be cell growth promoting factors were decreased immediately after LLL treatment, thereafter were acutely increased in cellular protein profile. 4. In ultrastructural changes of gingival fibroblasts treated by LLL, more prominent rough endoplasmic reticulum, mitochondrial hyperplasia/hypertrophy and increased extracellular fibrillar matrix were observed in comparison with control group under same experimental period.