• Journal of Periodontal and Implant Science
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• 제33권2호
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• pp.277-288
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• 2003

It has been suggested that increased number and activity of phagocytes in periodontitis lesion results in a high degree of reactive oxygen species (ROS) such as superoxide anion, hydrogen peroxide, nitric oxide and peroxynitrite. There are few reports on the relationship between ROS and MMPs expressions in gingival fibroblast. We studied to elucidate whether and how ROS, especially nitric oxide affects the MMP expression. Human gingival fibroblasts and HTl080 cells (human fibrosarcoma sell line as reference) were grown in DMEM supplemented with 10 mM HEPES, 50 mg/L gentamicin, and 10% heat inactivated fetal bovine serum with addition of various reactive oxygen species (ROS). Culture media conditioned by cells were examined by gelatin zymography. HT1080 cells expressed proMMP-2 and proMMP-9, but human gingival fibroblasts (HGF) produced only proMMP-2. Hydrogen peroxide upregulated MMP-9 expression in HT1080 cells, whereas in human gingival fibroblast SNP treatment showed marked increase in MMP-2 level compared to other ROS. These results suggest that the effects of ROS on MMPs expressions are cell-type specific. RT-PCR for MMP-2 and TIMP-2 m-RNA were performed using total RNA from cultured cells under the influence various kinase inhibitors. In HT1080 cells, treatment with FPTI III (Ras processing inhibitor) and LY294002 (PI3-kinase inhibitor) resulted in inhibition of MMP-2 and MMP-9 expressions, suggesting that Ras/P13-kinase pathway is important for MMPs expression in HT1080 cells. In gingival fibroblasts, treatment with FPTI III and PDTC (NF-kB inhibitor) showed marked decrease in MMP-2 regardless of the of SNP , suggesting that Ras/NF-kB could be the key pathway for NO-induced MMP-2 expression in gingival fibroblasts. This study showed that ROS, especially nitric oxide, could be the critical mediator of periodontal disease progression through control of MMP-2 expression in gingival fibroblasts possibly via Ras/NF-kB pathway.

• 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.

• Journal of Periodontal and Implant Science
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• 제26권3호
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• pp.681-688
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• 1996

Periodontal regeneration requires the migration and proliferation of gingival fibroblasts and periodontal ligament cells. These cellular events are influenced and regulated by growth factors and some drugs. The purpose of this study is to examine the effect of Rhizoma coptidis and Centella asiatica extracts on human gingival fibroblasts. Gingival fibroblasts were primarily cultured from extracted premolar with non-periodontal diseases. Cells were cultured with ${\alpha}-MEM$ at $37^{\circ}C$, 5% $CO_2$, 100% humidity incubator for 2 or 3 days, as a measure of cell proliferation potential, it was examined that the DNA synthesis using $[^3H]-thyrnidine$ incorporation, the cell numbers (with or without dye), and cell viabilities. Rhizoma coptidis is increased the proliferation of gingival fibroblasts at concentration of $10^{-9}g/ml$, but Centella asiatica is decreased the proliferation at all concentrations. This study demonstrated that Rhizoma coptidis is a potential mitogen for human gingival fibroblasts in vitro, and we can expect the usefulness of this drug in periodontal regeneration.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제27권5호
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• pp.424-430
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• 2005

The oral cavity is humid environment mainly due to the continuous salivary flow. The reaction of oral mucosa to fluid flow is important for homeostasis and pathogenesis. The objective of this study is the screening the change of gene expression after the application of fluid induced shear stress (FISS) on the gingival fibroblast using cDNA microarray assay. The immortalized human gingival fibroblasts were grown and FISS was applied using a cone viscometer at a rotational velocity of 40 rpm, respectively for periods of 2 and 4 hours. The synthesis of cDNA was done from the extracted total RNA and cDNA microarray assay was done subsequently. The genes that showed over 1.6 in the Cy3/Cy5 or the Cy5/Cy3 value were regarded as genes influenced significantly by the FISS application ion (/M/>0.7). The " RUNX-1" was increased its expression in 2 hours group and " RUN and SH3 domain containing 1" was increased its expression in 4 hours group. The "CC020415", "cyclin L1", "interferon regulatory factor1", "early growth response 1", "immediate early response 2", and "immediate early response 3" genes were increased their expression in 2 and 4 hours after FISS application. In conclusion, we could find many genes that were probably related to the FISS application. Interestingly, most of them were placed in similar molecular pathways and these findings improve the reliability of chip data and usefulness in overall screening. From this experiment, we could find many items for further study and it will make improvement in the understanding of intracellular events in response to FISS.

• Journal of Oral Medicine and Pain
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• 제20권1호
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• pp.53-65
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• 1995

The growth and synthetic activities of fibroblasts are regulated by cytokines and growth factors derived from activated inflammatory cells. Stimulatory effect of low level laser (LLL) radiation on wound healing seems to be in part due to direct stimulatory action on cell proliferation and synthetic activities of fibroblasts. Also indirect stimulatory effect on the fibroblast function through inflammatory or immune cells is another possible mechanism of biostimulatory action of LLL. This study was performed to determine the growth rate of human gingival fibroblasts obtained biopsy and culture, fibroblast cell line, and immune cell line by using $[^3H]-$ thymidine incorporation test. And gene expression pattern was also analyzed by using the DNA probe such as Hsp70, IL-1$\beta$, MIP-1$\alpha$ and actin cDNA. Proliferation rate of gingival fibroblast was increased by LLL irradiation, but no more effect was added by LPS or IL-1$\beta$ pretreatment Enhanced Hsp70 gene expression was found from gingival fibroblasts and fibroblast cell line COS by LLL irradiation., which was not more increased by LPS or IL-1$\beta$ pretreatment. LLL-irradiated promyelcytic cell line HL-60 and macrophage cell line RAW264.7 showed significant stimulatory effect of proliferation rate when compared with respective control. However there were no changes in growth rate of other immune cell tested in this study, such as B cell line WR19n.l and 230, helper T cell line Jurkat and Hut78, cytolytic T cell line CTLL-r8. By LLL-irradiation Hsp70 gene expression was increased in RAW246.7 and HL-60, not in CTLL-R8. And IL-1$\beta$ and MIP-1$\alpha$ gene expression were induced only from LLL-irradiated RAW264.7. These results led us to presume that LLL radiation may affect to the immune cells, especially to macrophage, through which it might promote wound healing process.

• Journal of Periodontal and Implant Science
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• 제23권2호
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• pp.228-242
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• 1993

Some therapeutic agents and medicaments may lead to pathologic changes in the gingival tissue, especially on the cultured human gingival fibroblasts. The purpose of this study was to investigate on the effect of diphenylhydantoin, retinoic acid to the human gingival fibroblast. Human gingival fibroblasts were cultured from the healthy gingiva of patients with orthodontic patients. Gingival fibroblasts were trypsinized and transferred to the wells of 96 well microtest plates. Next day, the medium was removed, fibroblasts were washed with HBSS, and the washed cells were cultured in growth medium added 5 or $10{\mu}g/ml$ of diphenylhydantoin, $10^{-5}M$, $10^{-6}M$ and $10^{-7}M$ of retinoic acid and glycyrrhetinic acid. The passage number of cultured fibroblasts were fifth and eighth. The cell morphology was examined by inverted microscope, the cell number was counted by hemocytometer, and cell activity was measured by the growth and proliferatiton assay using MTT assay. The fifth experiments were performed and statistical significance was measured by ANOVA. The cell morphology in the presence of retinoic acid was round irrespective of the presence of diphenylhydantoin and glycyrrhetinic acid(Fig 2-6). The proliferation of cells was not changed by diphenylhydantoin(Table 1). The cell activity showed the tendency to increase at the concentration of $10{\mu}$'/, of diphenylhydantoin (Table 2). The cell activity in the presence of retinoic acid glycyrrhetinic acid was decreased, and the increased cell activity by diphenylhydantoin was decreased by retinoic acid and glycyrrhetinic acid at the concentration of $10^{-7}M$(Table 3-5). These results suggested that the increased cell activity by diphenylhydantoin might be modulated by retinoic acid and glycyrrhetinic acid.

• Journal of Applied Biological Chemistry
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• 제48권3호
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• pp.138-143
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• 2005

Antibacterial, free radical scavenging, proliferative, and cytotoxicity effects of Doenjang extracts were examined. All samples except water extract showed strong antibacterial activity against oral bacteria, Streptococcus. pyogenes, S. mutants, S. sanguinis, S. sorbrinus, S. criceti, S. antinosus, S. gordonii, and Porphyromonas. gingivalis (MIC and MBC values: 0.08-1.25 and 0.16-2.50 mg/ml, respectively). DPPH method showed ethanol and ethyl acetate extracts are effective inhibitors of oral bacteria. Based on MTT assay, 24 h exposure to 0.31 mg/ml of all extracts, excepted water extract, resulted in strong cytotoxicity on KB cells. All extracts strongly inhibited human gingival fibroblast viability.

• 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.

• 대한치과교정학회지
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• 제22권1호
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• pp.273-283
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• 1992

To find out the differences between periodontal ligament cells (PDL cells) and gingival fibroblast cells (GFB cells), alkaline phosphatase, a marker enzyme for osteoblast, was used to measure the activities and $^{45}CaCl_2$ isotope was used to find out cellular and release of $^{45}Ca$, a requisite for bone formation,. PDL cells and GFB cells from 1 to 5 passages were also measured in alkaline phosphatase activity assay. By the use of above methods, followings were concluded that the PDL cells and the GFB cells have characteristics that are different from each other. In that PDL cells showed large amount of calcium uptake and large amount of calcium release in initial stage, they seem to possess characteristics which are similar to osteoblast-like cells. 1. The PDL cells, in contrast to the gingival fibroblast, showed exceedingly high alkaline phosphatase activity which was highest at the second passage, decreasing thereon. But gingival fibroblasts cells showed no distinct differences in alkaline phosphatase activity as the passage were elapsed. 2. For both PDL cells and GF cells, the $^{45}Ca$ uptake was greatest at 2 hours period. The PDL cells showed higher measuring than GFB cells through out the whole time period. 3. Whereas the GFB cells showed slow increase of $^{45}Ca$ release as time relapsed, the PDL cells showed rapid increase of $^{45}Ca$ release.

• International Journal of Oral Biology
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• 제32권2호
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• pp.67-73
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• 2007

Periodontitis is a chronic infectious disease that leads to periodontal destruction, and is one of the major causes of tooth loss in humans. The osteoclast differentiation factor (ODF), which is also known as the receptor activator of the NF-kB ligand (RANKL), is a surface-associated ligand on bone marrow stromal cells and osteoblasts. RANKL activates its cognate receptor, RANK, on osteoclast progenitor cells, which leads to the differentiation of mononucleated precursor cells. Osteoprotegerin (OPG) is a decoy receptor that is released from stromal cells and osteoblasts to inhibit the interaction between RANKL and RANK. Although the precise mechanism of bone loss in periodontitis is unknown, the differentiation and activation of osteoclasts by OPG-ODF-RANK signaling might play the role in periodontal bone destruction. The relationship between the concentration of sex hormones and the expression of ODF and OPG was examined by treating human gingival fibroblasts and periodontal ligament cells with the normal serum concentration of estrogen or progesterone during menstruation or at menopause. The ODF/OPG relative ratio was elevated at the concentration observed during ovulation in human gingival fibroblasts and at the concentration observed between ovulation and menstruation in periodontal ligament cells treated with estrogen. However, the ratio was <1 at all concentrations in both cells treated with progesterone. In the case of menopause simulated by estrogen depletion, the ratio was <1 in human gingival fibroblasts but >1 in periodontal ligament cells.