• 한국융합학회논문지
• /
• 제9권12호
• /
• pp.81-88
• /
• 2018

사람치은섬유모세포(human gingival fibroblast, HGF)는 치은조직에 존재하는 주요한 세포의 형태 중 하나로 외부 자극에 반응하여 다양한 염증대사물질을 생산한다. 본 연구에서는 돌김에탄올추출물(PYEE)이 Porphyromonas gingivalis로부터 분리한 lipopolysaccharide로 염증이 유도된 HGF-1 cell에서 항염 효과를 보이는지 분석하고자 하였다. LPS-PG에 의해 과발현된 iNOS와 COX-2는 PYEE의 처리에 의해 농도 의존적으로 발현이 감소되었고, nuclear factor $(NF)-{\kappa}B$ 또한 동일한 양상으로 활성이 억제되었다. 신호전달물질 중 c-Jun $NH_2$-terminal kinase (JNK)의 인산화만이 PYEE에 의해 억제되었다. 그리고 항염 작용에 관여하는 것으로 알려진 2상 효소 중 하나인 NAD(P)H:quinone dehydrogenase (NQO)-1도 분석하였고, 이 효소는 PYEE의 처리에 의해 강하게 발현이 유도가 되었다. 결론적으로 돌김에탄올추출물은 치주질환 에방과 치료를 위한 후보물질로 활용 가능할 것으로 사료된다.

• Journal of Periodontal and Implant Science
• /
• 제28권1호
• /
• pp.133-143
• /
• 1998

Diabetes mellitus is a systemic disease with profound effects on oral health and periodontal wound healing. Uncontrolled diabetes adversely affects surgical wound healing and is often associated with abnormal proliferation of fibroblasts. Human gingival fibroblasts and PDL cells were chosen because they are intimately involved in periodontal therapy and are important for the success of surgical procedure such as guided tissue regeneration. The aim of the present study was to elucidate whether cellular activity and collagen synthesis by glucose pre-treated human gingival fibroblasts and PDL cells are influenced by insulin, and whether healthy cells differ from glucose treated cells. Cells were cultured with DMEM at $37^{\circ}C$, 5% $CO_2$, 100% humidified incubator. To evaluate the effect of glucose on gingival fibroblasts and periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4\;cells/well$ culture plates and treated with 20 and 50mM of glucose for 5 days. Then MTT assay was carried out. To evaluate the effect of insulin on glucose-pretreated cells, the cells were seeded at a cell density of $1{\times}10^4\;cells/well$ culture plates and treated with 20 and 50mM of glucose for 5 days. After incubation, $10^3$, $10^4$ and $10^5mU/l$ of insulin were also added to the each well and incubated for 2 days, respectively. Then, MTT assay and collagen synthesis assay were carried out. The results indicate that cellular activity of gingival fibroblasts significantly increased by glucose while periodontal ligament cells were unaffected and cellular activity of gingival fibroblasts and periodontal ligament cells were unaffected by insulin. Collagen synthesis of gingival fibroblast with 20mM glucose and insulin unaffected, but 50mM glucose and insulin increased than control. Collagen synthesis of periodontal ligament cell with 20mM glucose and $10^5mU/l$ insulin significantly increased than other groups and 50mM glucose pretreated PDL cells significantly increased at $10^3mU/l$ insulin but decreased at $10^4mU/l$ insulin. Our findings indicated that these cell types differed in their growth response to glucose, and the increase in collagen synthesis was significantly raised at insulin level of $10^3mU/l$ in gingival fibroblasts and periodontal ligament cells except 20mM glucose pretreated periodontal ligament cells.

• Journal of Periodontal and Implant Science
• /
• 제31권3호
• /
• pp.597-610
• /
• 2001

Normal gingival fibroblasts functioning is fundamental for the maintenance of periodontal connective tissue as well as wound healing. Nicotine have been found to affect DNA synthesis and cell proliferation, which appear to depend on the type of cells. This in vitro study was done to determine the effects of nicotine, a major component of tobacco, on cell proliferation, viability, activity, cell cycle distribution, and expression of cell cycle regulatory proteins in human gingival fibroblasts. Nicotine has been tested for 2 days or 4 days in 5 different concentrations; $0.1{\mu}g/ml$; $1{\mu}g/ml$; $10{\mu}g/ml$; $100{\mu}g/ml$; $1000{\mu}g/ml$. To assess cell proliferation and viability, viable and non-viable cells were counted by hemocytometer; to evaluate cellular activity, MTT assay was employed; to analyze cell cycle distribution, fluorescent propidium iodide-DNA complex were measured using fluorocytometer; to determine the expression of cell cycle regulatory proteins, western blot analysis was performed. After 2 days and 4 days incubation respectively, at concentrations of $1{\mu}g/ml$ - $1000{\mu}g/ml$, nicotine significantly inhibited proliferation comparing to non-supplemented controls. The cell viability was significantly decreased after 2 days and 4 days at concentrations of $1{\mu}g/ml$ - $1000{\mu}g/ml$ and at $10{\mu}g/ml$ - $1000{\mu}g/ml$ respectively. After 2 days and 4 days, the cellular activity was significantly decreased at concentrations of $10{\mu}g/ml$ - $1000{\mu}g/ml$. Treatment with $100{\mu}g/ml$ nicotine for 48 hours caused an increase in the proportion of G1-phase cells (from 46.41% to 53.46%) and a decrease in the proportion of S-phase cells (from 17.80% to 14.27%). The levels of cyclin $D_1$ and CDK 4 proteins in nicotine-treated fibroblasts were lower than that of controls, whereas the levels of p16 and pRB were higher than that of controls. These results suggest that the decrease of cell proliferation and lengthened Gap phases (G1) by nicotine may due to the increased expression of p16 and pRB as well as decreased expression of cyclin $D_1$ and CDK 4 in human gingival fibroblasts.

• 대한치과보철학회지
• /
• 제45권3호
• /
• pp.375-381
• /
• 2007

Statement of problem. Prior to determining an optimal width of micropatterned grooves provided on titanium substrata, we have done a pilot study using surface topographies in combined microm and submicrom levels. Purpose. The purpose of this study was twofold 1) to assess the proliferation and 2) to analyze the expression of genes encoding the intracellular signaling proteins involved in cell-substratum adhesions and adhesion-dependent G1 phase cell cycle progression of human gingival fibroblasts plated on smooth and microgrooved/acid-etched titanium substrata. Material and methods. Three groups of titanium discs as NE0 (smooth Ti substrata), E15 (Ti substrata with microgrooves of $15{\mu}m$ of spacing and $3.5{\mu}m$ in depth and with further acidetching), and E30 (Ti substrata with microgrooves of $30{\mu}m$ spacing and $3.5{\mu}m$ in depth and with further acid-etching) served as the human gingival fibroblasts' substrata. Viability and proliferation of fibroblasts were determined using an XTT assay. Gene expressions of fibronectin, ${\alpha}5$ integrin, CDK4, and $p27^{kip}$ were analyzed in RT-PCR. Cell-substratum interactions were analyzed in SEM. Results. From the XTT assay at 24 h incubation, the mean optical density (OD) value of E15 was significantly greater than the values of E30 and NE0. At 48 and 96 h however, the mean OD values of E30 were significantly greater than the values of E15 and NE0. No differences in the expression of PCR transcripts at 96 h incubations were noted between groups, whereas at 48 h, an unexpected increase in the expression of all the transcripts were noted in E15 compared with other two groups. Fibroblasts were observed to orient and adhere inside the microgrooves. Conclusion. Micropatterned grooves and acid-etching on Ti substrata alter viability and gene expression of adhered human gingival fibroblasts.

• Journal of Periodontal and Implant Science
• /
• 제25권2호
• /
• pp.181-191
• /
• 1995

The ability of fibroblasts attach to teeth is of paramount imporance in re-establishing the lost connective tissue attachment after periodontal therapy. Tobacco contains a complex mixture of substances including nicotine. various nitrousamines, trace elements. and a variety of poorly characterized substances. The effects of nicotine on fibroblasts have reported an altered morphology and attachment of fibroblasts to substrates and disturbances in protein synthesis and secretion. This study examined the effect of nicotine, a major component of the particulate phase of tobacco smoke, on human gingival fibroblasts and periodontal ligament cells attachment to tissue culture surfaces and cellular activity of human gingival fibroblasts and periodontal ligament cells. Pooled human gingival fibroblasts made from extraction of 3rd molar were utilized between passage 4 and 5 and plated in 96 well plate at 20,000 cells per well. Cell number were determined using 3-(4,5-dimethylthiazole-2-y)2,5-diphenyltetrazolium bromide(MTI) , which is reflection of mitochondrial dehydrogenase activity. The concentration of nicotine used were 0.025, 0.05, 0.1, 0.2 and $0.4{\mu}M$, the average serum concentration for a smoker being approximately $0.1{\mu}M$. The results were as follows : 1. Attachment effects of nicotine on human gingival fibroblasts and periodontal ligament cells Excepts of $0.4{\mu}M$, the effects on attachment with increasing numbers of cells attaching with increasing nicotine concentrations, compared to control group. But over the 60min, return to control value. 2. The effect of cellular activity on human gingival fibroblasts and periodontal ligament cells. The cellular activity of human gingival fibroblasts and periodontal ligament cells were similar or decrease to control value at 1st incubation day. At 2nd incubation day, 0.05, 0.1, 0.2, $0.4{\mu}M$ concentrations were statistically different from control value on gingival fibroblasts group. But at 3rd incubation day, cellular activities of all experimental group were significantly decrease than control group.

• Journal of Periodontal and Implant Science
• /
• 제22권1호
• /
• pp.203.2-203.2
• /
• 1992

• Journal of Periodontal and Implant Science
• /
• 제25권2호
• /
• pp.279-289
• /
• 1995

The most important object of periodontal treatment is the perfect regeneration of destructed periodontal tissue. The healing of periodontal lesion is affected by several cells & factors, which result in formation of long juntional epithelium, root resorption, bony ankylosis or connective tissue attachment. And ideal healing is enhanced by epithilial exclusion or periodontal ligament cell activation. In this investigation, I studied the effect of Zizyphus Fructus extract which enhances biologic activity& collagen synthesis, on the chemotaxis & cell nature. The cells were obtained from interdental area & middle third area of the freshly extracted teeth for the orthodontic purpose. And they were fully incubated in${\alpha}-MEM$ solution containing $100{\mu]g/ml$ penicillin & $100{\mu]g/ml$ streptomycin followed by 6 generation incubation. The test cells were collected by trypsin-EDTA & centrifuge in the fully incubated cells, counted by Hernacyotmeter, incbated $5{\times}10^5/ml$ cells for 24 hours, re-incubated 24 hours in media containing natural extract and photographed. The cells were incubated for 4 hours in 48 well microchemotaxis chamber bisecting upper & lower chamber by 8ug/m pore polycarbonate membrane coating 5mg/ml gelatin solution. The migrated cells in microscope were counted, which meaned cell chemotaxis activity. The study had shown that the morphology of cell was spindle-shaped as the control group, and the subextract test groups were not significantly different. In gingival fibroblasts, the chemotaxis effect of PDGF was statistically significant compared to control group. The Zizyphus Fructus extract was more or less enhanced chemotaxis effect and in $1{\mu}g/ml$ concentration the chemotaxis effect was slightly elevated compared with $10{\mu}g/ml$ concentration. But, among the subextracts, it was not significantly defferent. In PDL cells, the chemotaxis effect of PDGF in statistically significant, and the zizyphus Fructus extract had shown the enhanced effect. The effect was slightly higher in $1{\mu}g/ml$ concentration than 10g/ml concentration,and no significance among the subextracts.

• Journal of Periodontal and Implant Science
• /
• 제25권3호
• /
• pp.478-486
• /
• 1995

Periodontal therapy for treatment of periodontitis involves the elimination of bacterial plaque and elimination of the anatomic defects by regenerative procedure. The purpose of this study was to evaluate on the biological effect of magnolia and Ginkgo biloba extract to the antimicrobial, antiinflammatory and cellular activity. Antimicrobial assay was performed with the diffusion method of the extract by measuring of growth inhibitory zone of B. cereus from blood agar plate. Effect of the extract to cellular activity of gingival fibroblast were examined using MTT method and measured the result with optical density on 570nm by ELISA reader. Inhibitory effects of $PGE_2$ production from gingival fibroblast was performed with the addition of $IL-l{\beta}$ and the extract to the well and examined to the product of $PGE_2$ from cell by ELISA reader. In vivo anti-inflammatory effect was performed with injection examined with clinically and histologically for their extent of mecrosis and inflammation. Antimicrobial activity of Magnolia extract showed significantly higher activity than that of control. However, GBE did not showed significant activity to compare with control, and mixture of Magnolia and GBE extract showed significantly higher activity than that of control. The effect of cellular activity to gingival fibroblast showed no significant differences of between control and Magnolia extract. However, GBE showed significantly higher rate of cellular activity to compare with control and even to PDGF-BB, and also showed same degree of cellular activity even though mixed with Magnolia extract. The inhibitory effect of $PGE_2$ production showed significantly reduction of $PGE_2$ production to compare with control, but its inhibitory effect was not much strong to compare with Indomethacin. In vivo, antiinflammatory effect of Magnolia extract to P. gingivalis injection of Hamster buccal check showed significantly reduction of inflammatory cell infiltration and tissue necrosis, but GBE showed no effect on the inhibition of inflammatory process. These results suggested that Magnolia and GBE extract possessed different kind of biological activity and also can be compensated on their activity with each other for elimination of bacterial plaque and anatonical defect.

• 구강회복응용과학지
• /
• 제38권2호
• /
• pp.90-96
• /
• 2022

목적: 이 연구의 목적은 표고 버섯 추출물의 치주 병원체에 대한 항균 활성과 인간 치은 섬유아세포에 대한 세포 독성을 조사하는 것입니다. 연구 재료 및 방법: 표고버섯을 물과 아세톤에 닮근 후, 상등액을 건조하여 추출물을 채취하였다. 추출물에 대한 치주 병원체의 감수성을 조사하였다. 인간 치은 섬유아세포에 추출물을 처리하고 CCK-8 용액을 이용하여 세포 생존율을 측정하였다. 결과: 표고버섯 물추출물은 2.5 mg/ml에서 치주병원체의 증식을 유의하게 감소시켰다(P < 0.05). 아세톤 추출물은 0.32 mg/ml에서 Porphyromonas gingivalis와 Tannerella forsythia의 성장을 유의하게 억제하였고, 0.64 mg/ml에서 Treponema denticola의 성장을 유의하게 억제하였다(P < 0.05). 추출물의 세포독성은 2.5 mg/ml의 농도에서 나타났으며, 1.25 mg/ml 농도의 추출물이 4시간 후부터 세포 생존률을 감소시키는 것으로 나타났다. 결론: 표고버섯 추출물은 치주염 유발 세균에 대한 항균 활성과 염증 완화 효과를 보였다. 따라서 추출물은 치주질환 예방 및 치료에 대한 후보물질이 될 수 있다.

• Journal of Oral Medicine and Pain
• /
• 제19권2호
• /
• pp.57-71
• /
• 1994

Gingival fibroblasts were cultured and subjected to the test of Northern blot analysis for the demonstration of various mRNA expression in response to the low level laser treatment. For duplication of in vivo. Wound healing process, fibroblasts were pretreated with proinflammatory cytokine interleukin-1$\beta$(IL-1$\beta$) or mitogenic substance phorbol 12-myristate 13-acetate(PMA) prior to laser irradiation. The results were as follows : 1. By the laser irradiation, the gene expression of collagen type I was markedly increased I n gingival fibroblasts, especially in the case of PMA pretreatment. The gene expression of collagen type IV, however, was not only affected by laser irradiation but also by chemical cell stimulation. 2. Oncogene v-myc expression was affected by both laser irradiation and IL-1$\beta$ or PMA stimulation, But v-fos gene expression was not detected in any case of this experimental system. 3. Heat shock gene(Hsp 70)was expressed constiutively, but slightly increased by laser irradiation. 4. mRNA of fibroblast growth factor(FGF) was induced by both laser irradiation and IL-1$\beta$ or PMA treatment.