• Restorative Dentistry and Endodontics
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• v.41 no.3
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• pp.167-175
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• 2016

Objectives: Endodontically treated teeth with insufficient tooth structure are often restored with esthetic restorations. This study evaluated the cytotoxicity and biological effects of yttria partially stabilized zirconia (Y-TZP) blocks in combination with several dental cements. Materials and Methods: Pairs of zirconia cylinders with medium alone or cemented with three types of dental cement including RelyX U200 (3M ESPE), FujiCEM 2 (GC), and Panavia F 2.0 (Kuraray) were incubated in medium for 14 days. The cytotoxicity of each supernatant was determined using 3-(4,5-dimethylthiazole- 2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays on L929 fibroblasts and MC3T3-E1 osteoblasts. The levels of interleukin-6 (IL-6) mRNA were evaluated by reverse transcription polymerase chain reaction (RT-PCR), and IL-6 protein was evaluated by enzyme-linked immunosorbent assays (ELISA). The data were analyzed using one-way ANOVA and Tukey post-hoc tests. A p < 0.05 was considered statistically significant. Results: The MTT assays showed that MC3T3-E1 osteoblasts were more susceptible to dental cements than L929 fibroblasts. The resin based dental cements increased IL-6 expression in L929 cells, but reduced IL-6 expression in MC3T3-E1 cells. Conclusions: Zirconia alone or blocks cemented with dental cement showed acceptable biocompatibilities. The results showed resin-modified glass-ionomer based cement less produced inflammatory cytokines than other self-adhesive resin-based cements. Furthermore, osteoblasts were more susceptible than fibroblasts to the biological effects of dental cement.

• Journal of Periodontal and Implant Science
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• v.29 no.3
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• pp.607-621
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• 1999

This study examined the human fibroblasts cell attachment to commercially pure titanium surface which had been instrumented by 3 types of periodontal instruments. Commercially pure titanium plates were uniformly scaled using plastic, stainless steel, titanium curette. these all experimental groups 65 undirectional strokes with the designated curettes. Alteration of the surfaces due to instrumentation was evaluated by Form Talysurf(R) and reported as Ra value(mean surface roughness). Then other experimental groups were immersed in a cell suspension of human gingival fibroblasts($1{\times}10^5$ cell/ml). After 3 days of culture, cell attachment and morphology was observed by SEM, and attached cell were counted by Hemocytometer. A significant difference in mean Ra value was observed for surface instrumented by metal curette compared to either control surface or surface instrumented by the plastic curette(P<0.01). No stastically significant difference was noted between control surface and those instrumented by the plastic curette. SEM observation showed that cell morphology and attachment to the commercially pure titanium plate was similar appearance on the all experimental groups. Experimental groups instrumented by titanium curette and stainless steel curette were more attached cell number than control group, but experimental group instrumented by plastic curette were similar with control groups(P<0.01). In summary, metal curette produced an significant alteration of the commercially pure titanium surface and more favorable surface topography for cell attachment. Otherwise plastic curette was insignificantly altered the commercially pure titanium surface(P<0.01).

• Journal of Oral Medicine and Pain
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• v.24 no.4
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• pp.361-374
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• 1999

F. nucleatum is a gram-negative obligate anaerobe which is the principal and most frequent cause of gingival inflammation and is the predominant pathogen isolated in subsequent periodontal breakdown. It is also one of the most numerous bacteria found in subgingival plaque samples from healthy sites; its numbers are about 10-fold greater in plaque from periodontally diseased sites. The purpose of this study is to examine the effects of outer membrane(OM), outer membrane vesicle(OMV), and lipopolysaccharide(LPS) from F. nucleatum ATCC 25586 strain on the growth of human gingival fibroblasts and HOS 941 cells, and on the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression of mouse splenocytes. For the examination of cytotoxic effects, $TNF-{\alpha}$ production and $TNF-{\alpha}$ mRNA expression, the MTT assay, the ELISA and the RT-PCR were performed, respectively. All extracts of F. nucleatum tested were cytotoxic to both of human gingival fibroblasts and HOS 941 cells, and the significant difference of cytotoxic activity among the extracts was not observed. In the effects of these extracts on the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression of mouse splenocytes, all extracts of F. nucleatum tested also stimulated the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression, but the effects of the OM extracts on the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression were higher than those of the OMV and the LPS extracts. The pattern of the $TNF-{\alpha}$ mRNA expression was similar to that of the $TNF-{\alpha}$ production. These results indicate that F. nucleatum seems to contribute to the pathogenesis of periodontal diseases at least by its cytotoxicity, directly and its $TNF-{\alpha}$ production, indirectly.

• Journal of Periodontal and Implant Science
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• v.24 no.2
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• pp.357-375
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• 1994

The purpose of this study was to investigate the differences of histochemical characteristics in inflammatory fibrous gingival hyperplasia (FGH), phenytoin-induced gingival hyperplasia(PIGH), idiopathic gingival hyperplasia(IDGH) and control groups (healthy and inflammatory gingiva) by immunohistochemical method with various antibodies and histomorphological analysis. In immunohistochemical finding, antibodies to inflammatory cells (T/B lymphocytes, macrophages, other monocytes), proliferating cell nuclear antigen(PCNA), epidermal growth factor(EGF), factor VIII, and type I collagen were used. 1. The inflammatory infiltrates in FGH were less than those in inflammatory gingiva. The composition of inflammatory cells of PIGH was similar with that of FGH. IDGH showed a similar histologic findings with healthy gingival tissue. 2. In FGH, the number of fibroblasts and newly-formed collagen fibers was increased. No significant increase of fibroblasts and the dense accumulation of thick collagen fibers were seen in PIGH. The increase of fibroblasts and the dense accumulation of thick collagen were seen in IDGH. 3. PCNA-positive cells were localized mainly in the area accumulated with inflammatory cells and blood vessels, significantly increased in all hyperplastic tissue groups, and distributed evenly in IDGH. 4. The distribution of EGF were not observed in healthy gingiva but detected locally in area with confluent blood vessels,without significant difference between the other tissue groups. This results suggest that inflammation plays a significant role in inducing hyperplastic change of gingival tissue. While in DIGH, drug itself as well as inflammation seems to attribute to hyperplastic change.

• Journal of Periodontal and Implant Science
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• v.23 no.1
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• pp.37-47
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• 1993

The native connective tissue attachment of the periodontium is known to be a complex consisting of gingival fibroblasts, periodontal ligament cells, gingival epithelial cells, cementum, alveolar bone and extensive extracellular matrix (collagen, glycoprotein and proteoglycans). The purpose of this study was to evaluate the effects of natural extracts on DNA, collagen and protein synthesis and inhibition of cytokine production in the gingival and periodontal ligament fibroblasts and gingival epithelial cells. Healthy gingival tissue was obtained from orthodontic treatment patients, and gingival epithelial cells, gingival fibroblasts and periodontal ligament cells were isolated and cultured from the samples. After treated with Ginseng protein, Pluronic F-68, Scutellariae Radix, centella asiatica, PDGF, IGF, DNA synthesis, total protein and collagen synthesis, and cytokine production of gingival epithelial cell, gingival fibroblast and periodontal ligamentcells were measured. MTT method for DNA synthesis, Peterkofsky and Dingerman method for total protein and collagen synthesis, and IL-1 ELISA kit for cytokine production were used. The proliferation of epithelial cells was enhanced in Centella asiatica, Ginseng protein, Pluronic F-68 and Scutellariae Radix. The activities of PDL cells were increased in PDGF, IGF, and Pluronic F-68. Higher collagen synthesis was observed in Scutellariae Radix and total protein synthesis was increased in Scutellariae Radix and PDGF. The inhibitory effects on IL-1, IL-6, $TNF-{\alpha}$ were observed in all exrracts.

• Biomedical Science Letters
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• v.10 no.2
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• pp.85-92
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• 2004

Immortalization of primary corneal cells has influence on pharmacy, medical and biological fields. Especially, investigation of immortalization mechanism using viral oncoproteins is useful for medical treatments, and these cell lines will be useful materials for toxic test of medical supplies and cell biological experiments. Rabbit corneal fibroblasts in culture undergo a finite number of divisions before they reach a terminally non-proliferating state known as replicative senescence. Therefore, we attempted to induce immortalization of rabbit corneal fibroblasts with SV 40 large T antigen. As a result of experiment, expression of SV 40 large T antigen was confirmed, and expression of proteins related to cell cycle repressor was decreased in the transfection group compared with non-transfection group. According to the results of cell cycle phase distribution test, SV 40 large T antigen-transfected cells had obtained higher proliferation rate than primary cells. It was confirmed that during induction of immortalization, SV 40 large T antigen was not able to increase telomerase activity. In conclusion, we made a rabbit corneal fibroblast cell line with SV40 large T antigen. This cell line will be useful for further studies of mammalian fibroblast biology, particularly with regard to angiogenesis and malignant transformation. In addition, this cell line offers opportunity for testing potential therapeutics and can be used for toxicity tests of materials or cosmetics. In the future, our cell line can potentially be utilized in a wide range of biology related fields.

• Biomedical Science Letters
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• v.12 no.2
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• pp.119-125
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• 2006

In relation to the tissue engineering, the cellular responses to the morphology of the scaffold surface are interesting topics. Human ligament fibroblasts (HLFs) were cultured on the micrpatterned silicone substrates subjected to cyclic stretch to simulate ligament motion. Groove and ridge width of silicone substrates was 10/50, 20/50, 20/10, and 20/20 ${\mu}m$ (groove/ridge ${\mu}m$) with a depth of $3{\mu}m$. Strain was applied over two days for 4 hours per day with a frequency of 0.5 Hz with the magnitudes of 4 or 8%. The purpose of this study was to evaluate ligament fibroblast alignment and cellular responses in relation to the pattern of microgrooved surface and stretching magnitude. Ligament fibroblasts in the microgrooved surface were elongated and aligned parallel to the microgrooves under no stretch. Uniaxial cyclic stretch induced cellular activities and their orientation rise in cellular response and the cells showed alignment and elongation perpendicular to the direction of the stretch. Biochemical analyses showed that the best cellular response was found on the $20/50{\mu}m$ under 8% stretch. The surface morphology and mechanical stretching were found to contribute to increase of proliferation, collagen production.

• Restorative Dentistry and Endodontics
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• v.24 no.1
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• pp.76-87
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• 1999

The effect of retrograde root-end filling materials(IRM, Super-EBA, Vitremer, MTA) on human periodontal ligament fibroblasts and osteoblasts was observed. The cell activities were evaluated by MTT assay, protein assay and alkaline phosphatase activity examination. The results as follows ; 1. After 24hrs culture, both E1 cells & PDL fibroblast adding root-end filling materials were suppressed cell activities but after 48hrs, cell activities were recovered. 2. Cell activity was lowest in Vitremer followed by IRM, MTA, Super-EBA. 3. Cell activity depression by Vitremer was not concerned with pH changes. 4. Protein synthesis by root-end filling materials were not significant difference in Both E1 cell & PDL fibroblasts but protein synthesis were a little increased by Super-EBA. 5. Alkaline phosphatase activity was increased in E1 cell by Super-EBA & MTA but was not significant differences in E1 cell by IRM & Vitremer. Alkaline phosphatase activity was a little depressed in PDL fibroblast by Vitremer. This findings suggest that these root-end filling materials may have important roles in promotion of PDL healing and consequently may be useful for clinical application in apical surgery.

• Restorative Dentistry and Endodontics
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• v.16 no.2
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• pp.99-117
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• 1991

The purpose of this study is to evaluate the effects of dentin bonding agents on the fibroblasts cultivated from human pulp tissue. The fibroblasts were cultured in DMEM/10%FBS medium. Whatman filter paper discs (6mm diameter) soaked with $2{\mu}l$ of dentin bonding agents were placed on a millipore filter (pore size $0.22{\mu}m$) contained in a 50mm Petri dish, and then, exposed for 10 minutes, 30 minutes, 1 hour, 6 hours, 12 hours, 24 hours, 4 days and 7 days in $37.^{\circ}C$, 5% $CO_2$ incubator. The results of the experiments were analyzed by counting the cells and measuring the protein contents at 1 day, 4 days and 7 days. The results of this study were as follows: l. CLEARFIL NEW BOND, LITE-FIL BOND, GLUMA 3 Primer and GLUMA 4 Sealer showed cytotoxicity compared to the control group in the cell counts and the protein contents. 2. GLUMA 4 Sealer showed the least cytotoxicity among the three dentin bonding agents. 3. The results of the cell count were simialr to the results of protein content measurement. 4. LITE-FIL BOND exhibited marked cytotoxicity during 1 day, but, the cytotoxicity was slightly reduced after 4 and 7 days. 5. In GLUMA 3 Primer group, it was not possible to count the cell numbers and measure the protein contents, but the degeneration of cells was observed under the inverted phase-contrast microscope.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.32 no.3
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• pp.272-278
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• 2006

Purpose: This study is designed to evaluate biocompatibility of three types of absorbable collagen GBR membrane in vitro. Material and Method: The human PDL fibroblasts culture was obtained through typical way and the cells used in the experiment was forth passage. The membranes examined were Experimental group A, B, C. All the 3-experimental groups were made of bovine pericardium and the membranes were excised into 5$\times$5mm respectively. The samples of the membranes were fixed on the 24-well plate with the double-sided adhesive tape. Then, 2ml of cell suspension which included $2{\times}10^4$cells was inoculated into the 24-well plate, and the cells were cultured for 1 week. Cellular viability and the alkaline phosphatase activity were measured with ELISA. The membranes in the culture were processed to examine with SEM. Results: The survival rate was highest in control and Experimental group A is the next, group B and group C in order of the value. The values are analyzed for statistical difference using Wilcoxon test. All the values of experimental groups are significantly lower than those of control, and the vaules among the experimental groups significantly differ from each other. Alkaline phosphatase level was identical order with the viable cell rate. SEM examination revealed that the PDL fibroblasts adherent on culture dish (control) and group A were spindle-shaped, but on group B and C, the cells were round-shaped without processes.