• Title/Summary/Keyword: Human Fibroblast

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Preparation and Biocompatibility of 6-amino-6-deoxychitosan for Immobilization of Epidermal Growth Factor (세포성장인자 고정화를 위한 6-amino-6-deoxychitosan의 제조와 생체적합성)

  • Son, Tae Il;Park, Se Hoon;Kang, Hahk Soo;Jang, Eui Chan
    • Applied Chemistry for Engineering
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    • v.16 no.2
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    • pp.226-230
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    • 2005
  • Chitosan derivatives, 6-amino-6-deoxychitosan (6A6DC) was successively prepared as a reactive carbohydrate for the stabilization of epidermal growth factor (EGF) by the reactions of chitosan with tosyl chloride, sodium azide, and lithium aluminum tetrahydride. The structure of 6A6DC was confirmed by EA, FT-IR, $^1H-NMR$ and $^{13}C\{^1H\}-NMR$. The degree of substitution (ds) of amino groups in 6A6DC was determined to be 0.7. 6A6DC did not show any cytotoxicity on the normal human dermal fibroblast (NHDF) proliferation at least in the range tested (0.3 g/mL 600 g/mL) and was considered as a suitable material for the stabilization of EGF against proteolytic degradation due to its non-cytotoxicity and high reactivity.

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
    • Journal of Korean society of Dental Hygiene
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    • v.19 no.2
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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.

Anti-inflammatory effect of Porphyra yezoensis ethanol extract through the inhibited NF-κB and JNK activation in LPS-PG stimulated HGF-1 cells (사람 치은섬유모세포에서 NF-κB와 JNK 활성 억제를 통한 돌김 에탄올 추출물의 항염증 효과)

  • Park, Chung-Mu;Yoon, Hyun-Seo
    • Journal of the Korea Convergence Society
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    • v.9 no.12
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    • pp.81-88
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    • 2018
  • Human gingival fibroblast (HGF) is the main cell type existed in periodontium and produces a variety of inflammatory mediators by external stimuli. In this study, the anti-inflammatory activity of Porphyra yezoensis ethanol extract (PYEE) on LPS-PG lipopolysaccharide from Porphyromonas gingivalis activated HGF-1 cell. Up-regulated iNOS and COX-2 expressions by LPS-PG were significantly attenuated by PYEE treatment in a dose-dependent manner. In addition, activated nuclear factor $(NF)-{\kappa}B$ was also dose-dependently inhibited by PYEE treatment. Among upstream signaling molecules, PYEE treatment inhibited phosphorylation of c-Jun $NH_2$-terminal kinase (JNK) but did not give any effect on other molecules. On the other hand, one of phase II enzymes, NAD(P)H:quinone dehydrogenase (NQO)-1, was analyzed due to its anti-inflammatory activity, which was upregulated by PYEE treatment. Consequently, PYEE could be candidates for the prevention and treatment of periodontal diseases.

A convergence study of cytotoxicity evaluation of adult dentifrices (성인용 치약의 세포독성 평가의 융합적 연구)

  • Choi, Yu-Ri;Shim, Youn-Soo;Jang, Sun-Ok
    • Journal of Digital Convergence
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    • v.17 no.3
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    • pp.305-312
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    • 2019
  • This study was conducted to effect the cell activity and cytotoxicity of dentifrice. For the study, 6 kinds of general dentifrice, 3 kind of whitening dentifrice, 2 kinds of natural dentifrice and SLS(sodium lauryl sulfate) of positive control group. Immortalized human gingiva fibroblast cell was used for the study, WST test for cell activity and Agar diffusion test for cytotoxicity. Agar diffusion test showed high cytotoxicity in general dentifrice test group and whitening dentifrice test group, but low cytotoxicity in natural dentifrice test group. As a result of cell nucleus staining, cell shape and nuclear activity showed that the highest activity in natural dentifrice group, followed by whitening dentifrice group and general dentifrice group. As a result of this study, the cytotoxicity of different ingredient and according to the use to dentifrice. As a result of this study, we confirm cytotoxicity of kind and components according to the purpose of using dentifrice. Therefore, it is necessary to indicate the detailed ingredients of dentifrice for the smart choice of consumers.

Activating transcription factor 4 aggravates angiotensin II-induced cell dysfunction in human vascular aortic smooth muscle cells via transcriptionally activating fibroblast growth factor 21

  • Tao, Ke;Li, Ming;Gu, Xuefeng;Wang, Ming;Qian, Tianwei;Hu, Lijun;Li, Jiang
    • The Korean Journal of Physiology and Pharmacology
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    • v.26 no.5
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    • pp.347-355
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    • 2022
  • Abdominal aortic aneurysm (AAA) is a life-threatening disorder worldwide. Fibroblast growth factor 21 (FGF21) was shown to display a high level in the plasma of patients with AAA; however, its detailed functions underlying AAA pathogenesis are unclear. An in vitro AAA model was established in human aortic vascular smooth muscle cells (HASMCs) by angiotensin II (Ang-II) stimulation. Cell counting kit-8, wound healing, and Transwell assays were utilized for measuring cell proliferation and migration. RT-qPCR was used for detecting mRNA expression of FGF21 and activating transcription factor 4 (ATF4). Western blotting was utilized for assessing protein levels of FGF21, ATF4, and markers for the contractile phenotype of HASMCs. ChIP and luciferase reporter assays were implemented for identifying the binding relation between AFT4 and FGF21 promoters. FGF21 and ATF4 were both upregulated in Ang-II-treated HASMCs. Knocking down FGF21 attenuated Ang-II-induced proliferation, migration, and phenotype switch of HASMCs. ATF4 activated FGF21 transcription by binding to its promoter. FGF21 overexpression reversed AFT4 silencing-mediated inhibition of cell proliferation, migration, and phenotype switch. ATF4 transcriptionally upregulates FGF21 to promote the proliferation, migration, and phenotype switch of Ang-II-treated HASMCs.

Effect of Korean red ginseng on collagen biosynthesis and MMP-I activity in human dermal fibroblast (홍삼성분이 섬유아세포의 콜라겐 생합성과 MMP-1 활성에 미치는 영향)

  • Kim, Na-Mi;Koo, Bon-Suk;Lee, Seong-Kye;Hwang, Eui-Il;So, Seung-Ho;Do, Jae-Ho
    • Journal of Ginseng Research
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    • v.31 no.2
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    • pp.86-92
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    • 2007
  • This study was carried out to develop health & functional food by using Korean red ginseng for prevention of skin wrinkles. Effects of Korean red ginseng on the collagen biosynthesis and inhibition of matrix metalloproteinase-I (MMP-1) activity in human dermal fibroblast were investigated. Crude saponin contents of Korean red ginseng water extract (WE), Korean red ginseng ethanol extracts (EE) and Korean Red ginseng purified extracts (PE) were 72 mg/g, 107 mg/g and 220 mg/g, respectively. We incubated human fibroblast cell with Korean red ginseng component by addition of l ${\mu}g/ml$, 5 ${\mu}g/ml$, 10 ${\mu}g/ml$. Amount of collagen biosynthesis was 1.86 ng/ml in control sample and 2.85 ng/ml, 2.05 ng/ml and 2.58 ng/ml in retinoic acid, EE and PE respectively. Furthermore, $ginsenoside-Rg_1$ and $ginsenoside-Rb_1$ were shown 2.01 ng/ml and 3.07 ng/ml. MMP-l activities of EE, PE, $ginsenoside-Rg_1$ and $ginsenoside-Rb_1$ were decreased to 92%, 94%, 91% and 78% respectively as compared with control. Cell proliferation were showed 84-96% in the Korean red ginseng components. The antioxidative SOD activities of the Korean red ginseng components were showed 28-69%, however it was lower than that of Vitamin C. From this results, we conclude that Korean red ginseng have a anti-wrinkle effect and $ginsenoside-Rb_1$ may be considered as a more effective component.

EFFECTS OF FLUORIDE MOUTHRINSE ON CELL ACTIVITY OF GINGIVAL FIBROBLASTS OF CHILDREN (불소양치용액이 소아 치은 섬유아세포의 세포활성에 미치는 영향에 관한 연구)

  • Lee, Dong-Hyun;Lee, Kwang-Hee
    • Journal of the korean academy of Pediatric Dentistry
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    • v.24 no.1
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    • pp.204-219
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    • 1997
  • The use of fluoride is one of the most effective methods for caries prevention. Fluoridation of public water supply has been recognized, for many years, as an effective way to reduce dental caries. The fluoride supplement has been recommended when the natural fluoride was unavailable or below the optimal range. However the mechanism of caries prevention by fluoride has not yet been clarified and it is well known that an overdose of fluoride results inacute and chronic toxicity, especially dental fluorosis. Fluoride mouthrinsing solution is widely used in dentistry due to its effectiveness in carrying anticariogenic action. Understanding the effects of fluoride mouthrinsing solution on human gingival fibroblasts will provide the safety rationale for its use during the caries preventive therapy. The purpose of this study was to evaluate the cytotoxic effect of fluoride mouthrinsing solution on the human gingival fibroblast in vitro. The human gingival fibroblasts were cultured from healthy gingiva on the extracted deciduous teeth of children. Cells were inoculated into a 24-well plate with $1{\times}10^4cells/well$ of medium at $37^{\circ}C$, 100% humidity, 5% $CO_2$ incubator for 24 hours. And the cells were counted by using the hemocytometer at each designed study. Human gingival fibroblasts were cultured in growth medium after one minute application range of 0.02%-0.2% NaF solution and 0.1% $SnF_2$ solution. The cells used in this study were between fifth to eighth passage number. The cell morphology was examined by inverted microscope and cell proliferation was measured by incorporating $[^3H]$-thymidine into DNA. DNA synthesis by human gingival fibroblasts was assessed by $[^3H]$-thymidine uptake assays while the cell activity was measured by MTT assay. Each concentrated fluoride mouthrinsing solution was estimated for its biocompatability with fibroblasts by the tissue culture technique. The results of this study were as follows : 1. It was observed that at 0.05%, 0.2% NaF mouthrinsing solution the cytoplasmic processes became globular. When 0.1% $SnF_2$ mouthrinsing solution was applied, the cytoplasmic process and cell morphology were disappeared. 2. DNA synthetic activity was reduced regardless of the concentration of the fluoride mouthrinsing solution. However, the result is statistically insignificant except 0.1% $SnF_2$ mouthrinsing solution(p<0.05). 3. Our results indicate that 0.02%, 0.05% concentrations of NaF mouthrinsing solution caused minimal cytotoxicity. But 0.2% NaF and 0.1% $SnF_2$ concentration were a significant difference between the cell activity in the experimental group and control group (p<0.05). 4. After appling 0.05% & 0.02% NaF fluoride mouthrinsing solution, cell activity was restored to the control groups level according to incubating time. The results suggest that direct exposure to fluoride solution inhibits gingival fibroblast activity. Therefore, for the most effective use of fluoride use, lowering the concentration of fluoride mouthrinsing is advisable because it maintains biocompatability and free ion in the oral fluid.

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Antioxidation and Inhibition of Matrix Metalloproteinase in UV-irradiated Human Dermal Fibroblast by Selaginella tamariscina (자외선이 조사된 사람 피부 섬유아세포에서 권백의 항산화와 MMP 발현에 미치는 영향에 대한 연구)

  • Sim, Gwan-Sub;Kim, Jin-Hwa;Kim, Jin-Hui;Lee, Bum-Chun;Pyo, Hyeong-Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.32 no.2 s.57
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    • pp.75-79
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    • 2006
  • In this study. we evaluated anti-aging activity of medical plants that protect the skin cell damage induced by UV irradiation. We have investigated diverse biological activities of Selaginella tamariscina as an anti-aging ingredient of cosmetics. S. tamariscina was found to show scavenging activities of radicals and reactive oxygen species (ROS) with the $IC_{50}$ values of $65.1{\mu}g/mL$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $40.9 {\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. For testing intracellular ROS scavenging activity, the cultured human dermal fibroblasts were analyzed by increase in dichlorofluorescein (DCF) fluorescence upon exposure to UVB $20 mJ/cm^2$ after treatment of S. tamariscina. UVA-induced MMP-1 protein and mRNA expression in human dermal fibroblasts were reduced in a dose-dependent manner by S. tamariscina. Moreover, S. tamariscina inhibited MMP-2 (gelatinase) activity in UVA-irradiated human dermal fibroblasts assayed by zymography and semi-quantitative RT-PCR. Taken together, these results suggest that S. tamariscina may act as an anti-aging agent by Increasing collagen and preventing the skin cell damage induced by UV irradiation, and imply that S. tamariscina nay be useful as a new ingredient for anti-aging cosmetics.

Generation and Characterization of a Monoclonal Antibody with Specificity for Mycoplasma arginini

  • Son, Yeon-Sung;Hong, Hyo-Jeong
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.547-552
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    • 2007
  • Previously, we generated monoclonal antibodies (MAbs) that bound to the surface of human embryonic stem cells (hESCs) in an attempt to discover new hESC-specific surface markers. In this study, MAb 47-235 (IgG1, ${\kappa}$) was selected for further characterization. The MAb bound to the surface of undifferentiated hESCs but did not bind to mouse ESCs or mouse embryonic fibroblast cells in flow cytometric analysis. The antibody immunoprecipitated a 47 kDa protein from the lysates of cell surface-biotinylated hESCs. Identification of the protein by quadrupole time of flight tandem mass spectrometry revealed that 47-235 binds to Ag 243-5 protein of Mycoplasma arginini. BM-Cyclin treatment of the hESCs that reacted with 47-235 resulted in loss of mycoplasma DNA and the reactivity to 47-235. Nevertheless, the hESCs that were reactive to 47-235 maintained self-renewal and pluripotency and thus could be differentiated into three embryonic germ layers.

Use of Moving Aeration Membrane Bioreactor for the Efficient Production of Tissue Type Plasminogen Activator in Serum Free Medium

  • Hyun Koo Kim;Moo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.1 no.1
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    • pp.32-35
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    • 1996
  • Amoving aeration-membrane (MAM) bioreactor was employed for the production of 2$\mu$g/mL of tissue type Plasminogen Activator (tPA)in serum free medium from normal human fibroblast cells. This system could maintain high cell density for long periods of steady state conditions in perfusion cultivation. Under normal operating condition, shear stress was as low as 0.65 dynes/$\textrm{cm}^2$ at the agitation speed of 80 rpm. Even though cell density gradually decreased with increasing agitation speed, tPA production increased linearly with increasing shear stress within a moderate range. This culture system allowed production of 2$\mu$g tPA/mL while maintaining a high cell denisty of 1.0$\times$107 viable cell/mL.

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