• Title/Summary/Keyword: fibroblasts

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Endothelial Cell Seeding Onto the Extracellular Matrix of Fibroblasts for the Developement of Small Diameter Polyurethane Vessel (소구경 폴리우레탄 인공혈관의 개발을 위한 세포외기질위의 혈관내피세포 배양)

  • 박동국;이윤신
    • Journal of Biomedical Engineering Research
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    • v.16 no.1
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    • pp.1-8
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    • 1995
  • A variety of experiments of endothelial cell seeding onto artificial vessels have been performed. To improve endothelialization, one or two components of the extracellular matrix (ECM) have been used as an underlying matrix. In this study, the whole ECM excreted from fibroblasts was used as an underlying matrix. Fetal human fibroblasts were cultured on a polyurethane (PU) sheet. After a conflu; ence was attained, the cytoskeleton and the nuclei of the fibroblast were destroyed using Triton-X. Mitomycin, or irradiation. Omental microvascular endothelial cells from adult human were seeded onto various substrates. After 12 days in culture, the cells were counted. It was observed that the ECM treated by irradiation had the highest cell number. In addition, the cells on this substrate exhibited the most typical endothelial cell morphology. For preliminary animal experiments the PU vessels (inner diameter, 1.5mm) coated with ECM were implanted in the infrarena] abdominal aorta of rat. After the vessels had been implanted for 5 weeks, it was found that the surface of the PU vessels was completely covered with endothelia] cells. In conclusion, we can state that the fibroblast-derived whole ECM makes a better underlying substrate for the endothelialization of small diameter artificial vessels.

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Extract of Ettlia sp. YC001 Exerts Photoprotective Effects against UVB Irradiation in Normal Human Dermal Fibroblasts

  • Lee, Jeong-Ju;An, Sungkwan;Kim, Ki Bbeum;Heo, Jina;Cho, Dae-Hyun;Oh, Hee-Mock;Kim, Hee-Sik;Bae, Seunghee
    • Journal of Microbiology and Biotechnology
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    • v.26 no.4
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    • pp.775-783
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    • 2016
  • The identification of novel reagents that exert a biological ultraviolet (UV)-protective effect in skin cells represents an important strategy for preventing UV-induced skin aging. To this end, we investigated the potential protective effects of Ettlia sp. YC001 extracts against UV-induced cellular damage in normal human dermal fibroblasts (NHDFs). We generated four different extracts from Ettlia sp. YC001, and found that they exhibit low cytotoxicity in NHDFs. The ethyl acetate extract of Ettlia sp. YC001 markedly decreased UVB-induced cytotoxicity. Additionally, the ethyl acetate extract significantly inhibited the production of hydrogen peroxide-induced reactive oxygen species. Moreover, it inhibited UVB-induced thymine dimers, as confirmed by luciferase assay and thymine dimer dot-blot assay. Thus, the study findings suggest Ettlia sp. YC001 extract as a novel photoprotective reagent on UVB-induced cell dysfunctions in NHDFs.

Sensitization to Doxorubicin by Inhibition of the Nrf2-Antioxidant System (Doxorubicin 매개 세포독성에 대한 Nrf2 경로의 역할)

  • Cho, Jeong-Min;Park, Hyun-M;Kwak, Mi-Kyoung
    • YAKHAK HOEJI
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    • v.52 no.1
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    • pp.67-72
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    • 2008
  • The use of doxorubicin, which is one of the most effective anticancer agents, is often limited by occurrence of acquired resistance in tumor cells. GSH has been shown to be involved in the development of this drug resistance. Transcription factor Nrf2 governs the expression of GSH synthesizing glutamylcysteine ligase (GCL), as well as multiple phase 2 detoxifying enzymes. Here we show that Nrf2 is one of factors determining doxorubicin sensitivity. Nrf2-deficient fibroblasts (murine embryonic fibroblasts, MEF) were more susceptible to doxorubicin mediated cell death than wild-type cells. Doxorubicin treatment elevated levels of Nrf2-regulated genes including NAD(P)H: quinone oxidoreductase (Nqo1) and GCL in wild-type fibroblasts, while no induction was observed in Nrf2-deficient cells. Doxorubicin resistance in human ovarian SK-OV cells was reversed by treatment with L-buthionine-sulfoxamine (BSO), which is depleting intracellular GSH. Finally, transfection of SK-OV cells with Nrf2 siRNA resulted in exacerbated cytotoxicity following doxorubicin treatment compared to scrambled RNA control. These results indicate that the Nrf2 pathway, which plays a protective role in normal cells, can be a potential target to control cancer cell resistance to anticancer agents.

Evaluation of Senescence Induced Prematurely by Stress. Application for cosmetic active ingredients

  • Morvan, Pierre-Yves;Romuald Vallee
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.285-290
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    • 2003
  • Living cells are continuously subject to all sorts of stress such as ultraviolet rays on skin cells. Tests made in various laboratories show that when young fibroblasts (Le. at the beginning of their proliferate life) were repeatedly put under stress at subletal doses, they acquired a phenotype similar to Senescence Induced Prematurely by Stress (SIPS). The work presented hereafter was made on a new model of senescence induced prematurely by stress from ultraviolet Brays (UVB). The human fibroblast model was put under repeated UVB stress, causing SIPS. Several ageing biomarkers were used in order to characterise the cells that underwent stress:. an increase in the proportion of positive cells with senescence associated $\beta$-galactosidase activity (SA $\beta$-gal) measured by a specific coloration,. the proportion in the different morphological stages that fibroblasts undergo during culture visualised by microscopic observation,. the expression of genes known for overexpressing during senescence, particularly fibronectin and apolipoprotein J, measured by Real Time-PCR,. the common deletion of 4,977 bp in mitochondrial DNA, evaluated by nested PCR. Studying the variation of these 4 biomarkers, we have evaluated the protective effect of a Laminaria digitata extract (LDE) that can be used as a natural active ingredient for anti-ageing cosmetics.

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Anticancer Effect of Ferulic Acid on Cultured Human Skin Melanoma Cells

  • Son, Byoung-Kwan;Choi, Yu-Sun;Sohn, Young-Woo
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.457-461
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    • 2006
  • It is demonstrated that phenolic compound has cytotoxic effect on cancer cells. Recently, ferulic acid is involved in anticancer activity by showing the decrease of cell viability in cancer cells. But, the anticancer mechanism of ferulic acid is left unknown. The purpose of this study was to examine the anticancer activity of ferulic acid on NIH3T3 fibroblasts and human skin melanoma cells (SK-MEL-3). The anticancer activity was measured by determining the cytotoxicy of ferulic acid on these cells. The cytotoxicity was measured by cell viability via XTT assay in these cells. In this study, ferulic acid decreased cell viability according to the dose-dependent manners after human skin melanoma cells were treated with various concentrations of ferulic acid for 48 hours. especially, ferulic acid remarkably decreased cell viability at a concentration of $120{\mu}M$ compared with control in human skin melanoma cells. While, ferulic acid did not show the significant decrease of cell viability at concentrations of $30{\sim}120{\mu}M$ in NIH3T3 fibroblasts. These results suggest that ferulic acid showed anticancer activity in cancer cells such as human skin melanoma cells by the decrease of cell viability significantly.

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Suppression of Cyclooxygenase-2 Expression of Skin Fibroblasts by Wogonin, a Plant Flavone from Scutellaria Radix

  • Chi, Yeon-Sook;Kim, Hyun-Pyo
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.96-96
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    • 2003
  • Previously, wogonin (5,7-dihydroxy-8-methoxyflavone) was found to suppress proinflammatory enzyme expression including cyclooxygenase-2 (COX-2), contributing to in vivo anti-inflammatory activity against skin inflammation. However, the detailed effect on each skin cell type has not been understood. Therefore, present investigation was carried out to find the effect of wogonin on inflammation-associated gene expression from skin fibroblasts in culture using reverse transcriptase-polymerase chain reaction. As a result, it was found that wogonin (10 - 100 ${\mu}$M) clearly down-regulated COX -2 expression from NIH/3T3 cells treated with 12-O-tetradecanoylphorbol 13-acetate, interleukin-1${\beta}$ or tumor necrosis factor-a. But, the expression levels of COX-1, interleukin-1${\beta}$ and fibronectin were not significantly affected. This finding was well correlated with significant reduction of prostaglandin E$_2$(PGE$_2$) production by wogonin. As a comparison, NS-398 (selective cyclooxygenase-2 inhibitor) did not suppress COX -2 expression and other gene levels, while PGE$_2$production was potently reduced at 0.1 - 10 ${\mu}$M. All these results suggest that COX -2 down-regulation of skin fibroblasts may be, at least in part, one of anti-inflammatory mechanisms of wogonin against skin inflammation.

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Biological Differences between Hanwoo longissimus dorsi and semimembranosus Muscles in Collagen Synthesis of Fibroblasts

  • Subramaniyan, Sivakumar Allur;Hwang, Inho
    • Food Science of Animal Resources
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    • v.37 no.3
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    • pp.392-401
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    • 2017
  • Variations in physical toughness between muscles and animals are a function of growth rate and extend of collagen type I and III. The current study was designed to investigate the ability of growth rate, collagen concentration, collagen synthesizing and degrading genes on two different fibroblast cells derived from Hanwoo m. longissimus dorsi (LD) and semimembranosus (SM) muscles. Fibroblast cell survival time was determined for understanding about the characteristics of proliferation rate between the two fibroblasts. We examined the collagen concentration and protein expression of collagen type I and III between the two fibroblasts. The mRNA expression of collagen synthesis and collagen degrading genes to elucidate the molecular mechanisms on toughness and tenderness through collagen production between the two fibroblast cells. From our results the growth rate, collagen content and protein expression of collagen type I and III were significantly higher in SM than LD muscle fibroblast. The mRNA expressions of collagen synthesized genes were increased whereas the collagen degrading genes were decreased in SM than LD muscle. Results from confocal microscopical investigation showed increased fluorescence of collagen type I and III appearing stronger in SM than LD muscle fibroblast. These results implied that the locomotion muscle had higher fibroblast growth rate, leads to produce more collagen, and cause tougher than positional muscle. This in vitro study mirrored that background toughness of various muscles in live animal is likely associated with fibroblast growth pattern, collagen synthesis and its gene expression.

Chronic Treatment of Ethanol Inhibits Proliferation of Normal Fibroblasts, but Not Oncogenic ras-Transformed Cells

  • Gu, Young-Hwa;Park, Mi-Sun;Jhun, Byung-H.
    • Biomolecules & Therapeutics
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    • v.6 no.4
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    • pp.345-350
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    • 1998
  • The adverse effects of ethanol on cell proliferation have been described for a variety of tissues and cells. In the present study, we investigated whether chronic ethanol intoxication impairs the cell proliferation and DNA synthesis induced by oncogenic $H-ras^{V12}$ - and $v-K-ras^{V12}$-transformed cells. Ethanol treatment inhibited the cell proliferation and the DNA synthesis of control parental fibroblasts in a time- and dose-dependent manner. In contrast, ethanol did not suppress the proliferation of either oncogenic $H-ras^{V12}$ - or $v-K-ras^{V12}$ -transformed fibroblasts. Microinjection of oncogenic $H-Ras^{V12}$ protein induces DNA synthesis and ethanol treatment did not interfere with the DNA synthesis. The antiproliferative toxicity of ethanol was rescued by antioxidants, such as N-acetylcysteine and 4-methlpyrazole. These results indicate that the antiproliferative action site of ethanol toxicity lies upstream or is independent of Ras and ethanol exerts its toxicity through a free radical formation.

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A Cytotoxic Comparison of Contact Lens Care Solutions on Cultured Mouse fibroblasts (콘택트렌즈 관리용액이 배양 생쥐 섬유모세포에 미치는 세포독성에 관한 연구)

  • Kim, Jae-Min
    • Journal of Korean Ophthalmic Optics Society
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    • v.1 no.2
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    • pp.13-18
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    • 1996
  • In this study, contact lens care solutions were compared to each other for cytotoxic effect on cultured mouse fibroblasts by the use of neutral red(NR) assay. This study tested the cytotoxicity of 12 cleaning solutions, 2 lubricants and 5 multipurpose solutions(MPS) at 1%, 2%, 3% and 5% concentrations. These solutions are manufactured in Korea and foreign countries. The relative cytotoxic comparison of these solutions showed that some of them are toxic, three of the cleaning solutions were especially highly toxic 10 cells, so most fibroblasts were dead at 1% concentration. The toxic cleaning solutions have more components than other solutions. But both lubricants and MPSs are non-cytotoxic 10 cells. Some of these solutions did not have any descriptions or indications. They are very dangerous to eyes. From this study, contact lens care solutions should be tested for cytotoxic effects.

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Glucosamine Inhibits Lipopolysaccharide-induced Inflammatory Responses in Human Periodontal Ligament Fibroblasts

  • Kim, Eun Dam;Park, Hyun-Jung;Baek, Jeong-Hwa
    • International Journal of Oral Biology
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    • v.39 no.4
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    • pp.221-228
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    • 2014
  • Glucosamine is commonly taken by the elderly without prescription as a nutritional supplement to attenuate the progression or symptoms of osteoarthritis. Previous studies demonstrated that glucosamine shows anti-inflammatory effects in tissues such as blood vessels and the heart. However, there have been few reports about the effects of glucosamine on oral inflammatory diseases. Therefore, in this study, the effects of glucosamine on lipopolysaccharide (LPS)-induced inflammatory responses were investigated using human periodontal ligament fibroblasts (HPDLFs). HPDLFs were incubated in the presence and absence of glucosamine (10 mM) for 24 h, followed by treatment with E. coli LPS (100 ng/ml) or vehicle. Quantitative RT-PCR and ELISA results showed that LPS exposure significantly increased the levels of IL-6 and IL-8 mRNA and protein, while the effect was significantly suppressed by glucosamine treatment. Glucosamine did not attenuate, but slightly increased, the LPS-induced activation of mitogen activated kinases (ERK, p38, JNK). However, it suppressed the LPS-induced increase in the DNA binding affinity and transcriptional activity of NF-${\kappa}B$. These results suggest that glucosamine exerts anti-inflammatory effects on HPDLFs exposed to LPS via inhibition of NF-${\kappa}B$ activity, necessitating further studies using animal periodontitis models.