• Title/Summary/Keyword: human gingival fibroblasts

검색결과 144건 처리시간 0.021초

Comet assay를 이용한 Ferric Sulfate의 유전자 독성에 대한 연구 (EVALUATION OF THE GENOTOXICITY OF FERRIC SULFATE BY COMET ASSAY)

  • 강호승;김신;정태성;박혜련
    • 대한소아치과학회지
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    • 제27권1호
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    • pp.77-84
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    • 2000
  • 치수절단술은 유치의 치수치료 방법 중 사용빈도가 높은 시술 중 하나로 치수절단 술식에 사용되는 약제는 치수나 주위조직에 무해하여야 하며, 감염이나 내흡수 등의 부작용이 없어야 한다. 본 연구는 임상에서 유치의 지혈적 치수절단 술식의 약제로 사용되는 ferric sulfate의 유전자 독성을 평가할 목적으로 human gingival fibroblast에 ferric sulfate를 다양한 농도와 접촉시간을 설정한 후 comet assay를 이용하여 유전자 독성을 평가하여 보았다. 그 결과는 다음과 같다. 1. 농도에 따른 세포의 유전자 손상정도의 변화는 ferric sulfate의 농도에 비례하여 유전자 손상이 증가하는 양상을 나타내었다. 2. 농도에 따른 세포의 유전자 손상정도는 0.1mM 이상의 농도에서 대조군과 유의한 차이를 나타내었다(p<0.05). 3. 시간경과에 따른 세포의 유전자 독성의 변화는 대조군과 유의한 차이를 보이지 않았다(p>0.05).

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마이크로그루브 상 인간치은섬유아세포의 유전자 발현 분석: DNA microarray 연구 (Regulation of human gingival fibroblast gene expression on microgrooves: A DNA microarray study)

  • 이경호;이성복;안수진;박수정;이석원
    • 대한치과보철학회지
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    • 제55권4호
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    • pp.361-371
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    • 2017
  • 목적: 마이크로그루브 상 인간치은섬유아세포의 유전자발현감식을 DNA microarray를 이용하여 연구하는 것이다. 재료 및 방법: Grade II 티타늄 시편을 이용하여 표면에 마이크로그루브(폭/깊이: $60{\mu}m/10{\mu}m$, E60/10)를 형성하고 불산으로 산에칭하여 실험군으로 사용하였다. 표면처리를 하지 않은 평활한 티타늄 표면(NE0)을 대조군으로 사용하였다. 실험군과 대조군에 인간치은섬유아세포를 배양한 후 total RNA를 추출하였다. Oligonucleotide microarray를 시행하여 실험군과 대조군 간 다양한 유전자 발현량의 변화를 확인하였다. KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis를 통해 DNA chip의 발현 결과를 mapping하여 실험 조건에 따른 유전자 발현량의 변화를 pathway 수준에서 파악하였다. 결과: E60/10 마이크로그루브 표면과 NE0 표면에 대한 유전자 발현량 비교분석 결과, NE0 표면에 비하여 E60/10 마이크로그루브 표면에서 1.5배 이상 유의한 발현 차이를 보인 유전자는 123개, 2배 이상 유의한 발현 차이를 보인 유전자는 19개였다. 실험 조건에 따른 유전자 발현량의 변화를 KEGG pathway analysis를 통하여 확인하였고, 다양한 유전자 발현 결과들 중 대표적인 세포접착, 증식, 활성 관련 세포신호전달을 규명하였다. 결론: 마이크로그루브 표면은 다양한 유전자 발현 변화를 유도하고 관련 세포신호 전달을 유도한다. 본 연구의 결과에 따라서, 마이크로그루브는 유전자 발현 변화 및 세포신호 전달 활성화 등을 통한 세포활성도 증진을 필요로 하는 다양한 생체재료들의 표면으로 사용될 수 있다.

유체에 의해 유발된 전단력이 치은 섬유아세포 유전자 발현 변화에 미치는 영향에 관한 연구 (GENE EXPRESSION AFTER THE APPLICATION OF THE FLUID-INDUCED SHEAR STRESS ON THE GINGIVAL FIBROBLAST)

  • 정미향;최제용;채창훈;김성곤;남동석
    • 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.

염산테트라싸이클린으로 처리한 치근면의 치은상피세포부착에 관한 연구 (Effect of tetracycline-HCl root conditioning on gingival epithelial cell attachment to root surface)

  • 황나영;박병기;김상목;국중기;박주철;김병옥
    • Journal of Periodontal and Implant Science
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    • 제32권1호
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    • pp.13-23
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    • 2002
  • The ultimate goal of periodontal therapy is directed to arresting the progression of the disease, and regenerating the fibrous attachment. In order to achieve such treatment aim, the plaque and calculus must be eliminated and the physiological conditions of the root surface must be changed to facilitate the attachment and migration of the new fibroblasts, The method of changing the proper root surface conditions to promote the healing of periodontal tissue involves mechanical procedures, such as scaling and root planing, and chemical procedures such as tetracycline-HCl. However, the formation of a long junctional epithelium was most frequently observed type of healing. Thus, the aim of this study was to examine in vitro the influence of surface conditioning of dentin by TC-HCl on human gingival epithelial cell attachment. Human gingival epithelial cells were obtained from healthy retromolar pad area(under the age 23 years). Seventy two teeth extracted from severe periodontitis were used as study material. To evaluate the epithelial cell attachment to dentin, the prepared specimen was divided to four groups. For the control group, only scaling and root planing were carried out, and for the test group, 1 to 3, the concentration of the TC-HCl was 50, 125 and 250mg/ml respectively. After cell cultivation time of 1-, 3-. 24 hour, for the indirect quantitative assessment of gingival epithelial cell attached to dentin sample, the absorbance of epithelial cell unattached to dentin was measured. The results were as follows; 1. There was no statistically significant difference between scaling and root planing group and TC-HCl 50mg/ml 125mg/ml and 250mg/ml group about absorbance of unattached epithelial cell to dentin sample(p>0.5). 2. As time passes, the absorbance of unattached gingival epithelial cell to dentin sample was decreased statistically significant(p<0.05). 3. There was no statistically significant difference among the TC-HCl group(p>0.05) We concluded that there was similar effect on gingival epithelial cell attachment between TC-HCl conditioning on root surface and only scaling and root planing treatment

Cyclosporin A-induced Gingival Overgrowth is Closely Associated with Regulation Collagen Synthesis by the Beta Subunit of Prolyl 4-hydroxylase and Collagen Degradation by Testican 1-mediated Matrix Metalloproteinase-2 Expression

  • Park, Seong-Hee;Kim, Jae-Yoen;Kim, Hyun-Jeong;Park, Kwang-Kyun;Cho, Kyoo-Sung;Choi, Seong-Ho;Chung, Won-Yoon
    • International Journal of Oral Biology
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    • 제33권4호
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    • pp.205-211
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    • 2008
  • Gingival overgrowth can cause dental occlusion and seriously interfere with mastication, speech, and dental hygiene. It is observed in 25 to 81% of renal transplant patients treated with cyclosporine A (CsA). CsA-induced gingival overgrowth (CIGO) is caused by quantitative alteration of the extracellular matrix components, particularly collagen. However, the molecular mechanisms involved in the pathogenesis of CIGO remain poorly understood, despite intense clinical and laboratory investigations. The aim of the present work is to identify differentially expressed genes closely associated with CIGO. Human gingival fibroblasts were isolated by primary explant culture of gingival tissues from five healthy subjects (HGFs) and two patients with the CIGO (CIGO-HGFs). The proliferative activity of CsA-treated HGFs and CIGO-HGFs was examined using the MTT assay. The identification of differentially expressed genes in CsA-treated CIGO-HGF was performed by differential display reverse transcriptase-polymerase chain reaction (RT-PCR) followed by DNA sequencing. CsA significantly increased the proliferation of two HGFs and two CIGO-HGFs, whereas three HGFs were not affected. Seven genes, including the beta subunit of prolyl 4-hydroxylase (P4HB) and testican 1, were upregulated by CsA in a highly proliferative CIGO-HGF. The increased P4HB and testican-1 mRNA levels were confirmed in CsA-treated CIGO-HGFs by semiquantitative RT-PCR. Furthermore, CsA increased type I collagen mRNA levels and suppressed MMP-2 mRNA levels, which are regulated by P4HB and testican-1, respectively. These results suggest that CsA may induce gingival overgrowth through the upregulation of P4HB and testican-1, resulting in the accumulation of extracellular matrix components.

저염식으로 유도한 Cyclosporine 신독성 백서에서의 치은 변화 (Comparative Study of Gingival Changes in Cyclosporine-Induced Nephrotoxicity with Normal and Low Salt Diet)

  • 임재계;김용진;박용훈
    • Journal of Periodontal and Implant Science
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    • 제30권2호
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    • pp.359-374
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    • 2000
  • Cyclosporine A(CsA) is a widely used immunosuppressant for transplant patients and is also used for the treatment of a wide variety of systemic diseases with immunologic disorders. However, its use is frequently limited because of complications such as nephrotoxicity or gingival hyperplasia. Although several hypotheses have been postulated for CsA-induced gingival hyperplasia, i.e. various cytokine effects of inflammatory cells, existence of plaque or CsA itself, but its pathogenesis is still unclear. For experimental chronic CsA toxicity, salt depletion has been shown to increased susceptibility of rodents to the effects of CsA, and this maneuver facilitates production of arteriolopathy and interstitial fibrosis in kidney that mimic the changes found in human. The purpose of this study was to evaluate pathogenesis of CsA-induced gingival hyperplasia by comparing changes between CsA administration groups of normal standard diet and those of low salt diet group. Specific pathogen-free, 20 to 25 days old(120 to 150 g), male Fisher-344 rats(KIST, Korea), 120 to 150g of body weight, were assigned to four groups of six animals each after one week of adaptation period for powder food. Group 1 received olive oil($300{\mu}l/g\;of\;diet$) with normal standard diet(0.4% of sodium)(NSD). Group 2 received CsA(Cypol-N, Jonggundang, Korea; $300{\mu}g/g\;of\;diet$) with normal standard diet(NSD+CsA). Group 3 received same amount of olive oil with low salt diet(0.05 % of sodium, Teklad Premier, U.S.A.)(LSD). Group 4 received same dose of CsA with low salt diet(LSD+CsA). Rats were pair fed and were sacrificed after six weeks. Renal histologic lesions associated with CsA, consisted of cortical interstitial fibrosis, tubular atrophy and hyalinization of arterioles and the impairment of renal function including increase of serum creatinine and decrease of glomerular filtration rate was more severe in low salt diet group. These were proved as the results of activated of renin-angiotensin system in the kidney by low salt condition. Meanwhile the degree of gingival hyperplasia at incisor and molar tooth was less severe in low salt diet group compared with normal sodium diet group. Hyperplastic gingiva showed mild epithelial hyperplasia and expanded underlyng stroma which consisted of matrix increasement, capillary proliferation and dilatation. While the number and the activation of fibroblasts were increased, inflammatory cells were rare in the stroma. The immunohistochemistry for TGF-${\beta}_1$ in the kidney and gingiva revealed stronger positive in LSD+CsA in kidney but in gingiva of NSD+CsA. These results suggested followings; Gingival hyperplasia can be developed without inflammatory cells infiltration and seemed not induced by CsA by itself. The major role for gingival hyperplasia by CsA would be the secondary effect of TGF-${\beta}$, which maybe upregulated by CsA administration. Low salt diet can attenuate this hyperplasia perhaps by decreasing the activation of $TGF-{\beta}$.

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Antimicrobial Effect of Ethanol Extract of Garcinia mangostana L. against Enterococcus faecalis Isolated from Human Oral Cavity

  • Park, Tae-Young;Lim, Yun Kyong;Lee, Dae Sung;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • 제43권3호
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    • pp.129-132
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    • 2018
  • Enterococcus faecalis is a major causative agent of endodontic treatment failure. The purpose of this study was to investigate bactericidal effects of ethanol extract of Garcinia mangostana L. (mangosteen extract) on five strains of E. faecalis that were isolated from human oral cavities. The bactericidal effects of mangosteen extract were assessed by measurement of minimum bactericidal concentration (MBC) value. The cytotoxicity of mangosteen extract on immortalized human gingival fibroblasts, hTERT-hNOF, was determined based on cell counting method. The data revealed the MBC value of mangosteen extract against the E. faecalis strains was $4{\mu}g/ml$. Additionally, the cell viability of mangosteen extract on hTERT-hNOF was 83.7-89.1% at the 1 to $16{\mu}g/ml$. These findings indicated that mangosteen extract could be used as a root canal cleaner during management of endodontic treatment failure caused by E. faecalis.

Tissue engineering of dental pulp on type I collagen

  • Lee, Gwang-Hee;Huh, Sung-Yoon;Park, Sang-Hyuk
    • Restorative Dentistry and Endodontics
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    • 제29권4호
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    • pp.370-377
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    • 2004
  • The purpose of this study was to regenerate human dental pulp tissues similar to native pulp tissues. Using the mixture of type I collagen solution, primary cells collected from the different tissues (pulp, gingiva, and skin) and NIH 3T3 ($1{\;}{\times}{\;}10^5{\;}cells/ml/well$) were cultured at 12-well plate at $37^{\circ}C$ for 14 days. Standardized photographs were taken with digital camera during 14 days and the diameter of the contracted collagen gel matrix was measured and statistically analyzed with student t-test. As one of the pulp tissue engineering, normal human dental pulp tissue and collagen gel matrix cultured with dental pulp cells for 14 days were fixed and stained with Hematoxyline & Eosin. According to this study, the results were as follows: 1. The contraction of collagen gel matrix cultured with pulp cells for 14 days was significantly higher than other fibroblasts (gingiva, skin) (p < 0.05), 2. The diameter of collagen gel matrix cultured with pulp cells was reduced to 70.4% after 7 days, and 57.1% after 14 days. 3. The collagen gel without any cells did not contract, whereas the collagen gel cultured with gingiva and skin showed mild contraction after 14 days (88.1% and 87.6% respectively). 4. The contraction of the collagen gel cultured with NIH 3T3 cells after 14 days was higher than those cultured with gingival and skin fibroblasts, but it was not statistically significant (72.1%, p > 0.05). 5. The collagen gel matrix cultured with pulp cells for 14 days showed similar shape with native pulp tissue without blood vessels. This approach may provide a means of engineering a variety of other oral tissue as well and these cell behaviors may provide information needed to establish pulp tissue engineering protocols.

Evaluation of physical property and cytotoxicity of resin infiltrant based on a triethylene glycol dimethacrylate (TEGDMA)

  • Min, Ji-Hyun;Roh, Ji-Yeon;Kim, Ki-Rim
    • 한국치위생학회지
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    • 제19권2호
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    • pp.173-181
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    • 2019
  • Objectives: The resin infiltration technique is a promising alternative therapy for arresting the early dental caries. However, there are very few reports on the safety and biocompatibility of this technique. We evaluated various properties of resin infiltrant (RI) based on a triethylene glycol dimethacrylate (TEGDMA).The water sorption (Wsp) and water solubility (Wsl) was assessed. Additionally, the cytotoxicity of RI against both animal and human fibroblast cell lines was investigated. Methods: The RI of the $Icon^{(R)}$, the first product developed for resin infiltration, is mainly composed of TEGDMA in the resin matrix. The Wsp and Wsl for the RI were measured in accordance with ISO 4049 specifications. Fourier-transform infrared spectroscopy (FTIR) was used for analyzing the polymerization before and after curing of RI. The cytotoxicity of RI against the mouse fibroblasts (L929) and human gingival fibroblasts (hTERT-hNOF) was evaluated using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and the data were analyzed using one-way analysis of variance. Results: Wsp and Wsl of the RI specimens were $53.37{\mu}g/mm^3$ and $10.6{\mu}g/mm^3$, respectively. FTIR analysis revealed a slightly higher degree of curing with longer irradiation time. The degree of conversion for RI was high (80.9%) after 40 seconds of light curing. There was a significant decrease in the viability of L929 and hTERT-hNOF cells at RI extraction solution concentrations above 50%, respectively, compared to that in the negative control (p< 0.05). Conclusions: Even though the RI exhibited positive effect on the early prevention of dental caries, the clinicians should also consider the toxicity of RI on periodontal tissues.

Effect of laser-dimpled titanium surfaces on attachment of epithelial-like cells and fibroblasts

  • Lee, Dong-Woon;Kim, Jae-Gu;Kim, Meyoung-Kon;Ansari, Sahar;Moshaverinia, Alireza;Choi, Seong-Ho;Ryu, Jae-Jun
    • The Journal of Advanced Prosthodontics
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    • 제7권2호
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    • pp.138-145
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    • 2015
  • PURPOSE. The objective of this study was to conduct an in vitro comparative evaluation of polished and laser-dimpled titanium (Ti) surfaces to determine whether either surface has an advantage in promoting the attachment of epithelial-like cells and fibroblast to Ti. MATERIALS AND METHODS. Forty-eight coin-shaped samples of commercially pure, grade 4 Ti plates were used in this study. These discs were cleaned to a surface roughness (Ra: roughness centerline average) of 180 nm by polishing and were divided into three groups: SM (n=16) had no dimples and served as the control, SM15 (n=16) had $5-{\mu}m$ dimples at $10-{\mu}m$ intervals, and SM30 (n=16) had $5-{\mu}m$ dimples at $25-{\mu}m$ intervals in a $2{\times}4mm^2$ area at the center of the disc. Human gingival squamous cell carcinoma cells (YD-38) and human lung fibroblasts (MRC-5) were cultured and used in cell proliferation assays, adhesion assays, immunofluorescent staining of adhesion proteins, and morphological analysis by SEM. The data were analyzed statistically to determine the significance of differences. RESULTS. The adhesion strength of epithelial cells was higher on Ti surfaces with $5-{\mu}m$ laser dimples than on polished Ti surfaces, while the adhesion of fibroblasts was not significantly changed by laser treatment of implant surfaces. However, epithelial cells and fibroblasts around the laser dimples appeared larger and showed increased expression of adhesion proteins. CONCLUSION. These findings demonstrate that laser dimpling may contribute to improving the peri-implant soft tissue barrier. This study provided helpful information for developing the transmucosal surface of the abutment.