• Title/Summary/Keyword: human fibroblast

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Effective Expression of Recombinant Baculovirus Vector Systems (재조합 베큘로바이러스벡터의 효과적 발현)

  • Kim, Ji-Young;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2014.10a
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    • pp.977-980
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    • 2014
  • A baculovirus vector systems including genes of polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD) were constructed. These recombinant baculovirus vector systems were transfected into human foreskin fibroblast cells and various tissues and investigated gene transfer and expression of these vector systems with control vectors. From the study, these recombinant baculovirus vector systems were more effective and safe than control vector in view of gene transfer and expression.

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Inhibitory Effect of Kaempferol on Apoptosis Induced by Phorbol Ester via the Reduction of ROS in Normal Human Dermal Fibroblast

  • Park, Su-Ji;Lee, Sei-Jung
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2020.10a
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    • pp.219-219
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    • 2020
  • Kaempferol (3,4',5,7-tetrahydroxyflavone), a flavonoid found in beans, broccoli, garlic, etc., has been used in natural medicine as an anti-inflammatory and antioxidant. This experiment was carried out to evaluate the anti-apoptotic effect of kaempferol in 12-O-tetradecanoylphorbol 13-acetate (TPA)-treated Normal Human Dermal Fibroblast (NHDF). Kaempferol inhibited the production of intracellular Reactive Oxygen Species (ROS) induced by TPA in NHDF. Kaempferol significantly blocks the phosphorylation of extracellular signal-regulated kinase responsible for the activation of nuclear factor-kappa B. In addition, kaempferol significantly attenuated the expression of Bax and cleaved caspase-3 as regulated by the phosphorylation of nuclear factor-kappa B during its blockage of TPA-induced apoptotic cell death. These findings suggest that kaempferol protects the apoptotic signaling pathway induced by TPA through modulating intracellular ROS in NHDF.

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Variable Effect of Estrogen on Fibroblast Proliferation and Collagen Synthesis by Gender and Age (에스트로겐이 진피섬유아세포의 증식 및 교원질합성에 미치는 영향의 다양성)

  • Shin, Seung Han;Won, Chang Hoon;Han, Seung Kyu;Kim, Woo Kyung
    • Archives of Plastic Surgery
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    • v.32 no.3
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    • pp.363-368
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    • 2005
  • It was assumed that the effect of estrogen on wound healing would be variable according to patient's gender and age since estrogen is a sex steroid. This study was designed to determine the variability of the effect of estrogen on proliferation of human dermal fibroblasts and collagen synthesis which are most important in wound healing considering patient's gender and age. Fibroblasts were isolated from the dermis of female patients in premenstrual, menstrual, or postmenopausal age group and that of male patients. The isolated fibroblasts were cultivated in the presence of estrogen($1.0{\mu}g/ml$). The cells were seeded at $5.0{\times}10^3cell/well$ in Dulbecco's Modified Eagle's Medium/Ham's F-12 nutrient including 5% fetal bovine serum in 96-well plates. The cells were incubated for 3 days. For fibroblast proliferation MTT assay method was used. To measure the production of collagen, the collagen type I carboxy- terminal propeptide enzyme immunoassay was carried out. Estrogen stimulated the proliferation of fibroblasts in female patients, but not in male patients. The greatest cell proliferation and collagen synthesis was seen at women in menstrual and postmenopausal age. These results demonstrated that effects of estrogen on dermal fibroblast proliferation and collagen synthesis were variable with gender and age.

Skin Regeneration, Anti-wrinkle, Whitening and Moisturizing Effects of Cheongsangbangpung-tang Aqueous Extracts with Cytotoxicity (청상방풍탕 열수 추출물의 피부재생, 주름개선, 미백, 보습 효과 및 세포독성 평가)

  • Woo, Chang-Yoon;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.30 no.2
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    • pp.49-70
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    • 2017
  • Objectives: The present study is to observe the skin-regeneration, anti-wrinkle, whitening and skin moisturizing effects of Cheongsangbangpung-tang (CSBPT) with cytotoxicity. Methods: In the present study, cytotoxicity of CSBPT lyophilized aqueous extracts (yield=18.71%) was experimented against human normal fibroblast cells and B16F10 murine melanoma cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT) assay, and skin regeneration and anti-wrinkle effects were also showed through the assay of collagen type I synthesis by an enzyme immunoassay (EIA) kit as comparing with transforming growth factor (TGF)-${\beta}1$, hyaluronidase, collagenase and matrix metalloproteinase (MMP)-1 inhibitory assays as comparing with oleanolic acid (OA), and elastase inhibitory effects as comparing with phosphoramidon disodium salt (PP). In addition, whitening effects of CSBPT were observed by tyrosinase inhibitory assay and melanin formation test in B16/F10 melanoma cells as comparing with arbutin, and skin moisturizing effects were measured through mouse skin water contents test, respectively. Results: No CSBPT treatment related cytotoxic effects were demonstrated against human normal fibroblast cells and B16/F10 murine melanoma cells. CSBPT concentration-dependent increased collagen type I synthesis at human normal fibroblast cells. It also effectively suspreessed hyaluronidase, collagenase, elastase and MMP-1 activities, which were enzymes that related to declining of ECM and formation of wrinkle. CSBPT supressed B16/F10 melanoma cells's melanin productions with tyrosinase activity, which was an enzyme connected with melanin formation, and dose-dependent and significant increases of skin water contents were detected in CSBPT treated mouse skin as compared with vehicle control skins. Conclusions: CSBPT showed favorable and enough skin regeneration, anti-wrinkle, whitening and skin moisturizing effects at least in a condition of this experiment. However, more detail mechanism and in vivo skin protective efficacy studies should be conducted in future with the screening of the biological active compounds in individual herbs of Cheongsangbangpung-tang.

THE EFFECTS OF GLYCYRRHETINIC ACID AND OLEANOLIC ACID TO CYCLOSPORINE A INDUCED CELL ACTIVITY OF CULTURED GINGIVAL FIBROBLASTS (Glycyrrhetinic acid와 oleanolic acid가 배양 치은 섬유모세포의 cyclosporine A 유도 세포활성에 미치는 영향)

  • Kim, Young-Wook;Kim, Jae-Hyun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.24 no.2
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    • pp.238-254
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    • 1994
  • Cyclosporine A is an immunosuppressant commonly used for patients receiving organ transplants. Gingival overgrowth is an adverse side-effect seen in about 8-26% of patients taking cyclosporine A which have been shown to increase the DNA synthesis of gingival fibroblast at the concentration of $10^{-9}g/ml$ in vitro. Glycyrrhetinic acid is the active pharmacological ingredients of licorice which exerts steroid-like action and anti-viral activity. Oleanolic acid, which were isolated from Glechoma hederacea, has been shown to act as inhibitors of tumor promotion in vivo and to be less cytotoxic retinoic acid. This study has been performed to evaluate the effects of glycyrrhetinic acid and oleanolic acid on cyclosporine A induced cell activity in vitro. Human gingival fibroblasts were isolated from explant cultures of healthy gingiva of orthodontic patients. Gingival fibroblasts were trypsinized and transferred to the walls of microtest plates. Fibroblasts were cultured in growth medium added $10^{-9}g/ml$ cyclosporineA and $50{\mu}l/ml$ lipopolysaccharides. Cells between the 4th and 6th transfer in culture were used for this study. The morphology of gingival fibroblst were examined by inverted microscope. The effects of cyclosporine A on the time course of DNA sythesis by human gingival fibroblasts were assessed by $[^3H]-thymidine$ uptake assays. Cyclosporine A was found to stimulate DNA synthesis of human gingival fibroblast at a concentration of $10^{-9}g/ml$. In the presence of lipopolysaccharide derived from Fusobacterium nucleatum, addition of cyclosporine A results in reversal of inhibition at the concentration which normally inhibits gingival fibroblast proliferation. The cell acitivities in the presence of glycyrrhetinic acid and oleanolic acid were decreased, and increased cell acitivities by cyclosporine A were decreased by glycyrrhetinic acid and oleanolic acid at the concentration of $200{\mu}g/ml$. These results suggested that the increased cell activities by cyclosporine A modulated by glycyrrhetinic acid and oleanolic acid.

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Production of Varicella-zoster Virus Using Human Lung Fibroblast Cells As Host Cells (인체 폐섬유아세포 배양에 의한 수두바이러스의 생산)

  • 김원배;박정극
    • KSBB Journal
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    • v.11 no.2
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    • pp.254-261
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    • 1996
  • Attenuated varicella-zoster virus (VZV) was propagated in human lung fibroblast (HLF) cells Among media tested in this work, DMEM was the best medium for the growth of HLF cells. Because HLF was a normal finite cell line, cell growth late was dependent on the age of HLF cells. When the population doubling level (PDL) was higher than 46, apoptosis of HLF cells started and cell growth rate decreased. The optimum temperature for the cell growth and virus propagation in the T-flask culture was $37^{\circ}C$. In a microcarrier culture system in which Cytodex-3 was used for the VZV propagation in spinner vessels, the yield of plaque forming cells was lower than that in the T-flask culture. The relatively high shear environment near microcarriers was thought to cause the low yield of VZV in the microcarrier culture system.

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Effect of Torilis Fructus on Procollagen Biosynthesis and Activity of Matrix Metalloproteinase-I(MMP-1) in Human Dermal Fibroblast (사상자(Torilis Fructus)가 섬유아세포의 Procollagen 생합성과 Matrix metalloproteinase-I(MMP-1)의 활성에 미치는 영향)

  • Koo, Bon-Suk;Hwang, Eui-Il;So, Seung-Ho;Lee, Seong-Kye;Han, Gyeong-Ho;Kim, Na-Mi
    • Korean Journal of Pharmacognosy
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    • v.38 no.4
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    • pp.349-353
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    • 2007
  • Skin wrinkle formations are associated with collagen synthesis and matrix metalloproteinase-I(MMP-1) activity. This study was carried out to find out skin wrinkle reducing components in Torilis Fructus. Torilis Fructus were extracted with 70% ethanol and the ethanol extracts were systematically fractionated with n-hexane, ethylacetate, n-butanol and distilled water. Among them, antiwrinkle component from n-hexane fraction was purified by several column chromatographies and HPLC, which identified as torilin by $^1H-NMR,\;^{13}C-NMR$ and ESI-MS. To determine cell viability, collagen biosynthesis and MMP-1 activity, human dermal fibroblast was treated with 1-5 ppm concentrations of Torilis Fructus extract fraction and torilin. Cell viability was showed 84-102% at all group treated with 1-5 ppm. Collagen synthesis was increased in all group, especially torilin-treated group was highest amount. Active forms of MMP-1 were decreased in all group. From these results, we consider that Torilis Fructus have several antiwrinkle components and torilin may be one of the effective components.

Anti-wrinkle Effect of N-Acetyl-D-glucosamine (NAG) (N-Acetyl-D-glucosamine (NAG)의 피부주름 개선 효과)

  • Kim Kwang Soo;Choi Gun Ho;Choi Jang Woo;Choi Jun Hak;Han Song Hee;Nam Sang Yun;Lee Seung Hwa
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.4 s.48
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    • pp.457-462
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    • 2004
  • Anti-wrinkle effect of N-Acetyl-D-glucosamine (NAG) was evaluated by collagen synthesis and proliferation of normal human fibroblast. NAG was obtained by purifying deacetylated chitin which can be derived from chitin-rich crab shell. We studied in in-vitro cultures of human normal fibroblast, whether synthesis of collagens and fibroblast growth activation in these cells can be enhanced in the presence of NAG. It 야d not show any adverse effects in human shin irritation patch test. In in-vivo mouse test, it showed anti-wrinkle effect in hairless mouse (6W/F). From the HPLC analysis, the stability of NAG in the cosmetics product could be maintained for a long time. These results demonstrated that NAS can be useful anti-wrinkle cosmetic ingredient.

A Study on the Viability of Human Dermal Fibroblast Cell by Media for Ni-Cr alloy elution (치과용 Ni-Cr합금 용출배지에 의한 인간 피부 섬유아세포 성장도 관찰 연구)

  • Kim, Kap-Jin;Choi, Sung-Min;Kim, Chi-Young
    • Journal of Technologic Dentistry
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    • v.31 no.3
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    • pp.21-26
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    • 2009
  • Purpose: Standards of alloy for porcelain fused to metal crown be classified by metallic factor and biological factor. Metallic factors consist of stability of alloy composition and mechanical strength and surface characteristics for chemical bond. Biological factors be considered properties of metallic elements and problems originated by toxicity and hypersensitive reaction. Alloys considered such controversial points are the most suitable alloy for dental instrument. Method: Alloys added Be and Nb using Ni-Cr alloy which has been widely used for dental instrument be selected and classified experimental group. Non-addition Be and Nb to Ni-Cr alloy classify control group and addition Be alloy is Be-experimental group, addition Nb alloy is Nb-experimental group. Specimens for cytotoxicity analysis gave effect to washing and sterilization. and then made an experiment on elution with cell medium after disinfection. It conducted specimens within cell medium with 24hours, 48hours, 72hours, respectively. It cultured human dermal fibroblast(HDF) using cell medium for cytotoxicity test and then investigated elution rate through spectroscopic analysis by MTT-assay. Result: As results of cytotoxicity test by MTT-assay, cultured cell rate of VII measured more low numerical value within elution medium for 24hours focused on control group. Also, cultured cell rate of K3 alloys observed low value for 48hours, 72hours than value of control group. Conclusion: According to final result that synthesize above results, Ni-Cr alloy added Be and Ni has little difference in Cytotoxicity by MTT-assay.

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Anti-Inflammatory and Antioxidative Effects of Gracilaria textorii Ethanol Extract in LPS-PG-Stimulated Human Gingival Fibroblast-1 Cells (사람 치은섬유모세포에서 잎꼬시래기 에탄올 추출물의 항염증 및 항산화 효과)

  • Park, Chungmu;Yoon, Hyunseo
    • Journal of The Korean Society of Integrative Medicine
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    • v.7 no.4
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    • pp.61-69
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    • 2019
  • Purpose : Human gingival fibroblast cell is one of the the main cell types in periodontal tissue, which they can show anti-inflammatory activity through the production of numerous lines of inflammatory mediators such as inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and interleukins. Porphyromonas gingivalis, one of the oral pathogens, has reported to play a critical role in the development of periodontal diseases. This study aimed to investigate anti-inflammatory and antioxidative activities of Gracilaria textorii ethanol extract (GTEE) in P. gingivalis derived lipopolysaccharide (LPS-PG) stimulated human gingival fibroblast (HGF)-1 cell line. Methods : In order to analyze anti-inflammatory and antioxidative activities of GTEE in HGF-1 cell line, NOS enzyme activity, expression levels of iNOS, COX-2, NAD(P)H quinone dehydrogenase (NQO)1 and their transcription factors were estimated by Griess reaction and western hybridization. Results : LPS-PG induced overexpression of iNOS and COX-2, which was significantly attenuated by GTEE treatment in a dose-dependent manner without any cytotoxicity. In addition, intracellular NOS activity was in accordance with the result of iNOS expression. Due to important role in the regulation of inflammatory responses, phosphorylated status of p65 and c-jun, each subunit of nuclear factor (NF)-κB and activator protein (AP)-1, was also dose-dependently ameliorated by GTEE treatment. One of phase II enzymes, NQO1, and its transcription factor, nuclear factor erythroid 2-related factor 2 (Nrf2), were analyzed since elevated phase II enzyme expression inhibited inflammatory response, which was significantly elevated by GTEE treatment in HGF-1 cell line. Conclusion : In conclusion, GTEE mitigated LPS-PG-stimulated inflammatory responses by attenuating NF-κB and AP-1 activation as well as accelerating NQO1 and Nrf2 expression in HGF-1 cell line. These results indicate that GTEE might be utilized a promising strategy for potential anti-inflammatory agent in periodontal diseases.