• 치위생과학회지
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• 제12권6호
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• pp.689-695
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• 2012

The aim of this study was to elucidate the effects of concentrated growth factors (CGFs) on human gingival fibroblasts in vitro. Blood was collected from three male volunteers (average age 27 years). CGFs were prepared using standard protocols. The CGF exudates were collected at the following culture time points: 1, 7, 14, and 21 days. The levels of platelet-derived growth factor BB (PDGF-BB) and transforming growth factor ${\beta}1$ (TGF-${\beta}1$) in CGFs were quantified. The CGF exudates were then used to culture human gingival fibroblasts. The biologic characteristics of these fibroblasts were analyzed in vitro for 21 days. Platelet-rich plasma released the highest amounts of TGF-${\beta}1$ and PDGF-BB on the first day. The level of TGF-${\beta}1$ had decreased slightly by day 7, although the difference compared to levels at day 1 was not statistically significant. However, by days 14 and 21, levels of TGF-${\beta}1$ had dropped significantly compared to day 1 levels. The levels of PDGF-BB at days 7, 14, and 21 did not differ significantly from that measured on day 1. CGFs maintained the release of autologous growth factors for a reasonable period of time (7 days for TGF-${\beta}1$ and 21 days for PDGF-BB). Gingival fibroblasts treated with CGF exudates collected at day 14 reached peak viability and synthesized type I collagen. Furthermore, the CGF exudates exerted positive effects on the proliferation and differentiation of these cells at days 1, 7, 14, and 21. The findings of this study suggest that treatment with CGFs represents a promising method of enhancing mucosal healing following surgical procedures.

• 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.

• 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.

• 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.

• 대한한의학회지
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• 제31권2호
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• pp.91-108
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• 2010

Background & Objective: The aim of this study was to investigate the effect of P. radix on the inflammatory related gene expression in IL-$1{\beta}$-stimulated primary human gingival fibroblast using Whole Transcript Sense Target (WT-ST). Method: Human gingival fibroblast was incubated with P. radix [100 or $200\;{\mu}g/ml$], and IL-$1{\beta}$ [$1ng/m{\ell}$] added an hour later. After 24h, total RNA was extracted using RNeasy Mini Kit and the whole gene expression patterns were performed using WT-ST Labeling $Assay^{(R)}$. Result: In the DEG results, 782 genes were up-regulated in the IL-$1{\beta}$-treated group as compared to control and among those, 43 genes were associated with inflammation. 981 genes were down-regulated after treatment with IL-$1{\beta}$ and of those 7 genes were associated with inflammation. 1439 genes were up-regulated after treatment with P. radix plus IL-$1{\beta}$-treated when compared to IL-$1{\beta}$-treated alone group and 1225 genes were down-regulated in the same condition. Among the down-regulated genes, 5 were associated with inflammation- and inhibitor genes such as GDF15 and LIF. In the analysis of the P. radix plus IL-$1{\beta}$-treated group, the most significant pathways were the cytokine-cytokine receptor interaction, toll-like receptor signaling, JAK-STAT signaling and tyrosine metabolism. The gene expression patterns in the P. radix $200{\mu}g/m{\ell}$ plus IL-$1{\beta}$-treated group appear to be more involved in the metabolism-related pathways than in the $100{\mu}g/m{\ell}$ plus IL-$1{\beta}$-treated group. Conclusion & Discussion: By microarray analysis of gene expression data, we are able to identify gene expression patterns associated with not only anti-inflammation effect but also transcription function of P. radix.

• Journal of Periodontal and Implant Science
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• 제26권4호
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• pp.841-858
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• 1996

Epidermal growth factor(EGF) is one of polypeptide growth factors. EGF has been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. The purposes of this study is to evaluate the effects of EGF on the human periodontal ligament cells and human gingival fibroblast cells that promote regeneration of periodntal tissue. The mitogenic effects of epidermal 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 prepared cells were the primary cultured gingival fibroblast and periodontal ligament cells from humans, the fourth or sixth subpassages were used in the experiments. Cells were seeded in DMEM containing 10% FBS. 1, 10, 50, 100, $200{\eta}g/ml$ and epidermal growth factor were added to the quiescent cells for 24 hours, 48 hours and 72 hours. They were labeled with $10\{mu}l/200{\mu}l$ 5-Bromo-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 were increased dose dependently by epidermal growth factor at 24 hours, 48 hours and 72 hours. The mitogenic effects were similar at the 24 and 48 hours of epidermal 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 were increased dose dependently by epidermal growth factor at 24 hours but the DNA synthetic activity decreased at $200{\eta}g/ml$ of each hour. Generally the maximum mitogenic effects were observed at the 48 hours application of epidermal growth factor. The DNA synthetic activity of human periodontal ligament cells generally decreased lower at 24, 72 hours than at 48 hours the application of epidermal 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 epidermal growth factor. In conclusion, epidermal 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.

• 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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• 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.

• 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.

• 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.