• 제목/요약/키워드: Human Fibroblast

검색결과 799건 처리시간 0.028초

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

  • 김지영;홍성갑
    • 한국정보통신학회:학술대회논문집
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    • 한국정보통신학회 2014년도 추계학술대회
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    • pp.977-980
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    • 2014
  • polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), protein transduction domain (PTD) 유전자가 포함된 재조합 베큘로바이러스를 구축하였다. 본 재조합 베큘로바이러스 시스템은 인간 섬유아세포와 여러 가지 조직에 감염하여 시험하였고 재조합된 유전자의 전달과 유전자 발현을 대조 벡터시스템과 비교하였다. 본 연구의 결과로 제작된 재조합 베큘로바이러스 시스템은 유전자의 전달과 발현에 있어서 대조 벡터시스템 보다 더욱 효과적이고 안전적이었다.

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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
    • 한국환경과학회:학술대회논문집
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    • 한국환경과학회 2020년도 정기학술대회 발표논문집
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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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에스트로겐이 진피섬유아세포의 증식 및 교원질합성에 미치는 영향의 다양성 (Variable Effect of Estrogen on Fibroblast Proliferation and Collagen Synthesis by Gender and Age)

  • 신승한;원창훈;한승규;김우경
    • Archives of Plastic Surgery
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    • 제32권3호
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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.

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

  • 우창윤;김동철
    • 대한한방부인과학회지
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    • 제30권2호
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    • pp.49-70
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    • 2017
  • 목 적: 본 연구는 한의학에서 면포질환에 빈번히 사용하는 대표적인 복합처방인 청상방풍탕의 열수 추출물 동결건조물의 피부 노화 개선 효과 평가를 위하여 피부재생, 주름개선, 미백 및 보습 효과를 각각 평가하였고, 기본적인 독성 평가의 일환으로 세포독성 또한 in vitro 방법으로 평가하였다. 방 법: 본 연구에서는 human normal fibroblast(CRL-2076) 세포 및 B16/F10 murine melanoma(CRL-6475) 세포에 대한 청상방풍탕 열수 추출 동결건조물(수율 18.71%)의 세포독성을 MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium Bromide) 방법을 통해 평가하였고, B16/F10 melanoma cells의 melanin 생성 억제 정도 및 melanin 합성 필수 효소인 tyrosinase 활성 억제와 arbutin을 비교 평가하여 미백효과를 평가하였다. 또한 피부 재생 및 주름 개선 효과를 transforming growth factor(TGF)-${\beta}1$와 fibroblast의 collagen type I 합성능을 비교, phosphoramidon disodium salt(PP)와 세포외 기질의 분해에 관여하는 elastase활성 억제를 비교, oleanolic acid(OA)와 hyaluronidase, collagenase 및 matrix metalloproteinase(MMP)-1 활성 억제를 비교하여 각각 평가하였고, 마우스 피부 수분함량 변화를 관찰하여 보습효과를 평가하였다. 결 과: 본 실험의 결과, 청상방풍탕은 human normal fibroblast 세포 및 B16/F10 murine melanoma 세포에서 최고 농도인 500 mg/ml까지 유의한 세포독성이 나타나지 않았고, fibroblast의 collagen type I 합성을 증가시켰다. 또한 세포외기질 파괴와 연관된 것으로 알려진 hyaluronidase, elastase, collagenase 및 MMP-1 활성을 각각 억제하였고 melanin의 생성에 관여하는 tyrosinase의 활성 및 B16/F10 melanoma 세포의 melanin 생성을 농도 의존적으로 차단함이 관찰되었다. 이와 더불어 정상매체 대조군에 비해 청상방풍탕 경구 투여군이 투여 용량 의존적으로 마우스 피부 수분 함량을 유의성 있게 증가시켰다. 결 론: 이상의 결과에서, 청상방풍탕은 세포 독성 없이 피부재생, 주름개선, 미백 및 보습 효과를 나타남이 관찰되어 차후 피부 개선 소재로서 그 가치가 높을 것으로 판단되나 금후 개별 구성 약재 각각에 대한 효능 및 생리활성을 나타내는 화학성분의 검색과 더불어 다양한 방면으로 기전적인 연구와 피부 보호 효과에 대한 in vivo평가를 체계적으로 수행해야 할 것으로 판단된다.

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

  • 김영욱;김재현;신형식
    • Journal of Periodontal and Implant Science
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    • 제24권2호
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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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인체 폐섬유아세포 배양에 의한 수두바이러스의 생산 (Production of Varicella-zoster Virus Using Human Lung Fibroblast Cells As Host Cells)

  • 김원배;박정극
    • KSBB Journal
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    • 제11권2호
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    • pp.254-261
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    • 1996
  • 수두 바이 러스(Varicella-zoster virus, VZV) 백신을 생산하기 위하여 약독화된 수두 바이러스 주인 Oka 주를 사용하여 정상세포인 인체 폐세 포(human lung fibroblast cells)로부터 수두 바이 러스 생산조건을 검토하였다. 숙주세포의 배양을 위 한 최적 성장배지로는 DMEM이 조사된 배지 중에 서는 가장 좋음을 알 수 있었으며 숙주세포의 성장율은 세포 age의 지표인 population doubling level (PDL) 이 46 이상인 경우 낮아지는 것무로 확인되 었으며 따라서 수두바이러스 생산을 위해서는 PDL 4 46 이하의 young cell을 사용해야 할 것으로 사료된 다 한편 수두바이러스 생산을 위한 최적 증식조건 으로는 배양온도 $37^{\circ}C$, 혈청농도 2%, MOl 1:5였다. 숙주세포를 대량배양하기 위한 기본조건을 검토 하기 위하여 미 럽담체 (Cytodex-3)를 이용한 spm­n ner flask culture를 시행하였다. 미럽담체에서의 숙 주세포 성장은 T -flask와 비하여 별다른 차이가 없었으나 VZV의 증식은 T-flask 배양에서보다 훨씬 낮았다. 이는 미립담체배양에서의 높은 전단응력 과 미립담체끼리의 영김현상 때문으로 생각된다.

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

  • 구본석;황의일;소승호;이성계;한경호;김나미
    • 생약학회지
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    • 제38권4호
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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.

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

  • 김광수;최근호;최장우;최준학;한송희;남상윤;이승화
    • 대한화장품학회지
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    • 제30권4호
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    • pp.457-462
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    • 2004
  • N-Acetyl-D-glucosamine (NAG)는 보습제로 사용되어지는 히아루론산의 구성물질인 뮤코 다당류의 일종이며, 특히 화장품으로서 응용은 보습제로서의 사용이 최초이다. 본 실험에서는 게나 새우의 껍데기에서 추출된 키틴을 탈아세틸화 하여 얻은 NAG를 화장품 원료로서 적용하고자 하였다. 현재 기능성 주름 원료로 알려진 레티놀(retinol)과 NAG를 비교하기 위하여 섬유아세포의 활성능력 및 콜라겐 생성촉진 효과를 비교 실험하였으며, 제형 내에서의 안정성을 위하여 HPLC로 역가를 측정하였다. 실험결과, 게의 껍질로부터 유도된 NAG는 피부에 자극을 전혀 주지 않으면서 섬유아세포의 세포활성 및 콜라겐의 생성을 촉진시키는 효과를 나타내었으며, 헤어리스 마우스를 대상으로 실험을 실시한 결과 피부층의 변화를 통하여 주름의 감소 효능을 볼 수 있었다.

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

  • 김갑진;최성민;김치영
    • 대한치과기공학회지
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    • 제31권3호
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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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사람 치은섬유모세포에서 잎꼬시래기 에탄올 추출물의 항염증 및 항산화 효과 (Anti-Inflammatory and Antioxidative Effects of Gracilaria textorii Ethanol Extract in LPS-PG-Stimulated Human Gingival Fibroblast-1 Cells)

  • 박충무;윤현서
    • 대한통합의학회지
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    • 제7권4호
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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.