• The Journal of Advanced Prosthodontics
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• 제5권3호
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• pp.341-350
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• 2013

PURPOSE. To evaluate adherence of human gingival fibroblasts (HGFs) to transmucosal abutment of dental implant with different surface conditions with time and to investigate the roles of focal adhesion linker proteins (FALPs) involved in HGFs adhesion to abutment surfaces. MATERIALS AND METHODS. Morphologies of cultured HGFs on titanium and ceramic discs with different surface were observed by scanning electron microscopy. Biocompatibility and focal adhesion were evaluated by ultrasonic wave application and cell viability assay. FALPs expression levels were assessed by RT-PCR and western blot. RESULTS. There seemed to be little difference in biocompatibility and adhesion strength of HGFs depending on the surface conditions and materials. In all experimental groups, the number of cells remaining on the disc surface after ultrasonic wave application increased more than 2 times at 3 days after seeding compared to 1-day cultured cells and this continued until 7 days of culture. FALPs expression levels, especially of vinculin and paxillin, also increased in 5-day cultured cells compared to 1-day cultured fibroblasts on the disc surface. CONCLUSION. These results might suggest that the strength of adhesion of fibroblasts to transmucosal abutment surfaces increases with time and it seemed to be related to expressions of FALPs.

• 치과방사선
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• 제28권1호
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• pp.59-71
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• 1998

The radiation-induced apoptosis was studied for two human cancer cell lines (KB cells, RPMI 2650 cells) and the human gingival fibroblast cell line (HGF-1 cells). The single irradiation of 2, 10, 20Gy was done with 241.5 cGylmin dose rate using the /sup 137/Cs MK cell irradiator. The cells were stained with propidium iodide and examined under the fluoro-microscope and assayed with the flow cytometry a day after irradiation. Also, the LDH assay was done to determine the amount of necrotic cells. The obtained results were as follows: 1. On the fluoro-microscope, many fragmented nuclei were detected in the KB, RPMI 2650, and HGF-1 cells after irradiation. 2. On the DNA content histogram obtained from the flow cytometry, the percentages of the pre-Gl peak of the control and 2, 10 and 20Gy irradiation group were 4.5, 55.0, 52.3, and 66.6％ on KB cells, 2.7, 3.3, 31.8, and 32.6％ on RPMI 2650 cells and 2.8, 21.8, 30.4, and 40.2％ on HGF-1 cells respectively. 3. The number of Gl-stage cells was abruptly decreased after 2Gy irradiation on KB cells and 10Gy irradiation on RPMI 2650 cells, But there was a slight decrease without regard to irradiation dose on HGF-1 cells. 4. There was no significantly different absorbance in extracellular LDH assay along the experimental cell lines.

• 한국융합학회논문지
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• 제9권12호
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• pp.81-88
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• 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
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• 제38권sup2호
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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.

• 대한치과보철학회지
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• 제45권3호
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• pp.375-381
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• 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
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• 제29권4호
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• pp.821-838
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• 1999

On the basis of the evidences that electrical stimulation could enhance proliferation and differentiation of bone cells and promote healing and regeneration of bone, this study was performed to investigate the effects of electrical stimulation on human periodontal ligament cells and gingival fibroblasts in vitro, which also have important roles in regeneration of periodontium, and to evaluate the potential of clinical application of electrical stimulation. Human periodontal ligament cells and gingival fibroblasts were primarily cultured from the root surface of extracted premolar and the adjacent gingiva without periodontal diseases. In control group, the cells ($5{\times}10^4$ cells/ml)were incubated only in Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum. In test groups, electrical stimulation was given at the current intensity of $0.25{\mu]A$(test group 1), $1.0{\mu}A$(test group 2), and $2.5{\mu}A$(test group 3) for 12 hours to the same culture media with the control group. After 12 hour exposure of electrical stimulation, the cells were incubated for 2 and a half days(60 hours), and then each group of cells was analyzed for cell proliferation, protein level, and activity of alkaline phosphatase. The results were as follows ; 1. The Rate of cell proliferation of every test group increased significantly in both periodontal ligament cells and gingival fibroblasts, and in periodontal ligament cells, test group 3 showed significantly increased proliferation compared to the other test groups(p<0.05). 2. In the protein levels, neither periodontal ligament cell nor gingival fibroblast showed statistically significant differences between control and test groups. 3. The activity of alkaline phosphatase in periodontal ligament cells increased significantly in all test groups(p<0.05), but there were no significant differences between 3 test groups. In gingival fibroblasts, the activity of alkaline phosphatase increased significantly only in test group 3(p<0.05). From the above results, it is concluded that electrical stimulation may have beneficial effects on the regeneration of destructed periodontal tissue in regard of the stimulation of periodontal ligament cells and gingival fibroblasts as well as electrically stimulated bone formation that has been known, and that electrical stimulation may have the potential of clinical application.

• Journal of Periodontal and Implant Science
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• 제26권1호
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• pp.133-142
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• 1996

Ten intrabony defects in 10 patients were treated by flap surgery including root surface debridement and placement of an expanded polytetrafluoroethylene(ePTFE) membrane. The membranes were removed after 4-6 weeks. This study was performed to examine the retrived ePTFE membrane by scanning electron microscopy(SEM) for bacterial contamination and adherent connective tissue elements, and to compare it with clinical conditions. The cervical portion of the membrane, which in most cases had become partially exposed to the oral cavity, had a bacterial deposit. Small bacterial colonies and a scatter of single cells in some instances extended into the apical portion of the membrane. Fibroblast-like cells, erythrocytes and fibrous structures were seen in the apical portion of the membrane. Outer surface of membrane tends to more bacterial contamination than inner surface(p<0.01), and upper portions more than lower portions(P<0.01). Comparison of ultrastructural findings and clinical conditions revealed that extent of bacterial contamination of the membrane correlated with gingival inflammation and extent of membrane exposure, but it was not significant statistically. The results suggested that gingival inflammation and membrane exposure affect periodontal regeneration by the use of ePTFE membrane.

• Journal of Microbiology and Biotechnology
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• 제28권8호
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• pp.1270-1281
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• 2018

Periodontal disease is triggered by the host immune response to pathogens in the microbial biofilm. Worsening of periodontal disease destroys the tooth-supporting tissues and alveolar bone. As oral inflammation can induce systemic diseases in humans, it is important to prevent periodontal disease. In this study, we demonstrated that Curcuma xanthorrhiza supercritical extract (CXS) and its active compound, xanthorrhizol (XAN), exhibit anti-inflammatory effects on lipopolysaccharide (LPS)-treated human gingival fibroblast-1 cells and anti-osteoclastic effects on receptor activator of nuclear factor kappa B ligand (RANKL)-treated RAW264.7 cells. LPS-upregulated inflammatory factors, such as nuclear factor kappa B p65 and $interleukin-1{\beta}$, were prominently reduced by CXS and XAN. In addition, RANKL-induced osteoclastic factors, such as nuclear factor of activated T-cells c1, tartrate-resistant acid phosphatase, and cathepsin K, were decreased in the presence of CXS and XAN. CXS and XAN inhibited the mitogen-activated protein kinase (MAPK)/activator protein-1 (AP-1) signaling pathway. Collectively, these results provide evidence that CXS and XAN suppress LPS-induced inflammation and RANKL-induced osteoclastogenesis by suppressing the MAPK/AP-1 pathway.

• Journal of Korean Dental Science
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• 제7권2호
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• pp.80-86
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• 2014

Purpose: Current common method of cytotoxicity evaluation for elastomeric impression materials use animal based cell lines, which the clinical relevance has been often questioned. Hence, the purpose of this study was to examine the difference in results with both human based and animal based fibroblast cell line. Materials and Methods: Three types of fibroblast cells were used in this study; conventional mouse fibroblasts of L929, human gingival fibroblasts (HGF-1), and immortalized human oral fibrobalsts (hTERT-hNOF). Test on extract and test by direct contact using different commercially available elastomeric impression materials were carried out according to the international standards. Result: There was significant difference in cell viability between types of fibroblasts cell used, where HGF-1 showed highest cell viability and L929 the lowest. Conclusion: Within the limitation of this study, careful consideration must be given when selecting the cells and interpreting the results for cytotoxicity evaluation of elastomeric impression materials, where use of human based cell lines such as hTERT-hNOF would be appropriate for both ease of cytotoxicity test and clinical relevance.

• Journal of Periodontal and Implant Science
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• 제29권3호
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• pp.621-636
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• 1999

When mucoperiosteal flaps are positioned and sutured to desirable position, the wound contains several interface between tissues which differ fundamentally in composition & biological reaction. Thus the C-T surface of the flap will, on one hand, oppose another vascularized surface, and on the other, the avascular dental material for example, when root resoptions, fractured root, endodontic perforation, deep root carious lesions were filled with amalgam, glass ionomer, resin etc. Recently, a number of case report described the successful treatment of a subgingival root lesion with restorative material & free gingival graft, open flap surgery, but more objective research was needed . Most of study on restorative materials were concerned for cytotoxicity not for actual healing event on that materials and its influencing factors such as biocompatibility, surface wettability, surface topography . The aim of this in vitro study was to evaluate the effect of amalgam, resin modified glass ionomer, composite resin per se, and their surface roughness on the growth of human gingival fibroblast. The cells were obtained and placed on culture flask and incubated for 3 days with the prepared test materials. Then count the attached cell number with hemocytometer,(n=12) and 2 samples were examined with SEM about attachment cell morphology . Another 4 samples were evaluated on their surface roughness with Talysurf and average surface roughness value(Ra) were obtained. Statistical difference in attached cell number, roughness value were analyzed using ANOVA. The number of attached cell was as follows, for root dentin specimen 16.7${\pm}$4.41, resin modified glass ionomer 14.0${\pm}$4.15, resin 8.13${\pm}$3.63, amalgam 0.72${\pm}$3.33(${\times}10^3$). Between root dentin and resin-modified glass ionomer, no significant difference was observed, but resin, amalgam showed a significant less cell numbers than for root dentin, resin modified glass ionomer cement. SEM examination expressed many cell surface attachment apparatus in root dentin and resin modified glass ionomer specimens. For resin specimen, cell attachment was observed but exposed less appratus. The average surface roughness value are following results. Dentin specimen 0.6972${\pm}$ 0.104, resin modified glass ionomer 0.0822${\pm}$0.009, resin 0.0875${\pm}$0.005, amalgam 4.2145${\pm}$0.985(${\mu}m$). Between root dentin, resin-modified glass ionomer, and resin, no significant difference was observed, but amalgam showed a significant more rough surface than other groups. When evlauated the interrelationship between cell attachment and surface roughness, therefore, there was weak reverse correlation.(pearson correlation : - 0.593) These results suggest that resin modified glass ionomer have the favorable healing potential when used for subgingival restoration. And for relationship between cell attachment and surface characteristics, further investigations were needed.