• Title/Summary/Keyword: Human skin fibroblast

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Modulation of Large Conductance $Ca^{2+}-activated$ $K^+4$ Channel of Skin Fibroblast (CRL-1474) by Cyclic Nucleotides

  • Yun, Ji-Hyun;Kim, Seung-Tae;Bang, Hyo-Weon
    • The Korean Journal of Physiology and Pharmacology
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    • v.9 no.2
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    • pp.131-135
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    • 2005
  • Potassium channels in human skin fibroblast have been studied as a possible site of Alzheimer disease pathogenesis. Fibroblasts in Alzheimer disease show alterations in signal transduction pathway such as changes in $Ca^{2+}$ homeostasis and/or $Ca^{2+}-activated$ kinases, phosphatidylinositol cascade, protein kinase C activity, cAMP levels and absence of specific $K^+$ channel. However, little is known so far about electrophysiological and pharmacological characteristics of large-conductance $Ca^{2+}$-activated $K^+$ ($BK_{Ca}$) channel in human fibroblast (CRL-1474). In the present study, we found Iberiotoxin- and TEA-sensitive outward rectifying oscillatory current with whole-cell recordings. Single channel analysis showed large conductance $K^{+}$ channels (106 pS of chord conductance at +40 mV in physiological $K^+$ gradient). The 106 pS channels were activated by membrane potential and $[Ca^{2+}]_i$, consistent with the known properties of $BK_{Ca}$ channels. $BK_{Ca}$ channels in CRL-1474 were positively regulated by adenylate cyclase activator ($10{\mu}M$ forskolin), 8-Br-cyclic AMP ($300{\mu}M$) or 8-Br-cyclic GMP ($300{\mu}M$). These results suggest that human skin fibroblasts (CR-1474) have typical $BK_{Ca}$ channel and this channel could be modulated by c-AMP and c-GMP. The electrophysiological characteristics of fibroblasts might be used as the diagnostic clues for Alzheimer disease.

Effect of Kimchi Extracts to Reactive Oxygen species in Skin Cell Cytotoxicity (김치 추출물의 활성산소에 대한 피부세포 독성 완화효과)

  • 류승희;전영수;권명자;문정원;이영순;문갑순
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.5
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    • pp.814-821
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    • 1997
  • Kimchi is composed of many ingredients such as Chinese cabbage, garlic, ginger, and red pepper and fermented fish extract. Some of them were known to have antioxidative activities due to their scavenging effect against reactive oxygen species(ROS). To study the health effects of kimchi on human skin cells, keratinocyte(A431, epidermoid carcinoma, human) and fibroblast(CCD-986SK, normal control, human) were cultured in oxidative stress condition provoked by paraquat, a superoxide anion generator, and hydrogen peroxide in the absence and presence of kimchi extract. The survival rate of keratinocyte was greatly reduced when exposed over 1mM concentration of hydrogen peroxide($H_{2}O_{2}$), but cytotoxicity of $H_{2}O_{2}$ was significantly reduced by kimchi extracts on cells. Especially 2 week-fermented kimchi decreased remarkably the cytotoxicity by $H_{2}O_{2}$ to keratinocyte cells. Over 1mM of paraquat concentration showed strong cell toxicity on keratinocyte, but the extracts from kimchi fermented for 1, 2 and 3 weeks showed protective effects in order. Fibroblast cells were significantly affected by $H_{2}O_{2}$ as were keratinocyte cells. Although almost all extacts of kimchi of different fermentation periods showed protective effect against cell killing at 0.5mM concentration of $H_{2}O_{2}$ week-fermented kimchi extract showed the strongest protective effect on fibroblast cells treated with 1mM $H_{2}O_{2}$ for either 1 day or 4 days. However most of kimchi extracts showed weak preventive effect or no effect on oxidative stress produced by paraquat. In conclusion, 2 week-fermented kimchi extract seems to have the best potential in preventing skin cells against oxidative damage which might be related to their scavenging effects of kimchi components produced during their fermentation process.

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In vitro Effects of the Complex of Korean Citrus junos and Medicinal Herbs on the Cell Growth in CCD-986sk Human Fibroblast Cell Line

  • Park, Kap-Joo
    • Korean Journal of Environmental Biology
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    • v.22 no.1
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    • pp.83-88
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    • 2004
  • In order to investigate whether or not CCD-980sk cell line can be affected by Korean Citrus junos and medicinal herbs, we examined the MTT assay when we treated Korean Citrus junos and medicinal herbs in CCD-986sk human fibroblast cell line. The samples that added complex of grinded extracts of Korean Citrus junos and boiling-water extracts from Korean medicinal herbs were tested toy cell proliferation activity by means of a modification of the MTT assay. Among mixture of Citron 3 (Citron 3, less mellowed citron which was ripened for three months) and boiling-water extracts, the group Citron 3+Phellinus linteus showed significantly strong cell proliferation activity. And among mixture of Citron 4 (Citron 4, completely mellowed citron which was ripened for four months) and boiling-water extracts, the group Citron 4+Cordyceps militaris and Citron 4+Phellinus linteus showed significantly strong cell proliferation activity, respectively. These results suggest that complex of Korean Citrus junos and medicinal herbs could be an excellent candidate toy protection of human skin aging.

The Protective Effect of Spirulina-derived Phycocyanin on Dermal Fibroblasts Induced by UV Rays (자외선으로 손상을 유도한 피부섬유아세포에서 스피룰리나 유래 피코시아닌의 보호 효과)

  • Yang, Jae Chan
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.5
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    • pp.1249-1254
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    • 2021
  • UV induce oxidative stress and increase matrix metalloproteinase (MMP) expression, resulting in skin aging. Thus, preventing skin damage from ultraviolet B (UVB)-induced skin damage can attenuate skin aging. Spirulina is comprised of prokaryotes a powerful antioxidant. This study aimed to investigate the photoprotective effects of spirulina-derived phycocyanin (PC) against UVB radiation using human skin fibroblast. As a results, PC showed no toxicity at concentrations of 5-40 ㎍/mL in terms of fibroblast viability. Survival rate of UVB-irradiated fibroblast incresased to 73.5% from 50.5% with PC treatment. UVB treatment increased MMP-1 and MMP-9 expression whereas PC treatment decreased it. The results indicate that PC might reduce or prevent skin aging by reducing UVB irradiation-induced skin wrinkles and free radicals.

Screening System Establishment for Potential Anti-wrinkle Agents Using Human Fibroblast Elastase (엘라스타제를 이용한 주름개선 후보물질 검색 시스템의 구축)

  • Oh, Mi-Hee;Lee, Ju-Eun;Kim, Su-Yeon;Kim, So-Young;Park, Kyoung-Chan;Yun, Hye-Young;Baek, Kwang-Jin;Kwon, Nyoun-Soo;Kim, Dong-Seok
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.1
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    • pp.19-25
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    • 2009
  • It has been reported that not only collagen but also elastin contribute to inhibit skin wrinkle formation. Ultraviolet (UV) radiation induces photo-aging on human skin. Because UV radiation increases elastase activity, it is thought that increased elastase activity could be the major reason for skin elasticity reduction and wrinkle formation by UV. In the present study to standardize elastase activity measuring system, purified elastases from porcine pancrease and human neutrophil, and cell extracts of normal human primary fibroblasts, 3T3 mouse fibroblasts, and CCD-25Sk human fibroblasts were used as various enzyme sources. Furthermore, elastase activities were compared according to concentrations of enzyme and substrate and incubation time. Phosphoramidon was used as a positive control to inhibit elastase activities of normal human primary fibroblasts and CCD-25Sk fibroblasts. However, it had no influence on the activity of porcine pancreatic elastase. Therefore, it is suggested that elastase used for testing anti-wrinkle agents should be selected carefully.

Fermented Acanthopanax koreanum Root Extract Reduces UVB- and H2O2-Induced Senescence in Human Skin Fibroblast Cells

  • Park, Min-Ja;Bae, Young-Seuk
    • Journal of Microbiology and Biotechnology
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    • v.26 no.7
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    • pp.1224-1233
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    • 2016
  • The present study assessed the effects of an aqueous extract of Acanthopanax koreanum root (AE) and of AE following fermentation by lactic acid bacteria (Lactobacillus plantarum and Bifidobacterium bifidum) (AEF) on human skin fibroblast HS68 cells exposed to ultraviolet B (UVB) irradiation and oxidative stress. AEF effectively antagonized the senescence-associated β-galactosidase staining and upregulation of p53 and p21Cip1/WAF1 induced by UVB or H2O2 treatment in HS68 cells. It also exhibited excellent antioxidant activities in radical scavenging assays and reduced the intracellular level of reactive oxygen species induced by UVB or H2O2 treatment. The antioxidant and antisenescent activities of AEF were greater than those of nonfermented A. koreanum extract. AEF significantly repressed the UVB- or H2O2-induced activities of matrix metalloproteinase (MMP)-1 and -3, overexpression of MMP-1, and nuclear factor κB (NF-κB) activation. This repression of NF-κB activation and MMP-1 overexpression was attenuated by a mitogen-activated protein kinase activator, suggesting that this AEF activity was dependent on this signaling pathway. Taken together, these data indicated that AEF-mediated antioxidant and anti-photoaging activities may produce anti-wrinkle effects on human skin.

Application of Human Dermal Fibroblast and Keratinocyte on Allogenic Dermis(AlloDerm®) (동종진피에 사람진피 섬유모세포와 각질세포를 적용한 인공피부의 실험적 제작)

  • Oh, Jung Chul;Lim, Yeung Kook;Jeong, Jae Ho
    • Archives of Plastic Surgery
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    • v.33 no.5
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    • pp.601-605
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    • 2006
  • Purpose: Large skin defect by various causes, should be covered by autologous skin graft. But, the donor site of autologous skin graft is limited and leaves permanent donor scar and contracture. There have been our trial to engineer artificial skin using allogenic dermis (AlloDerm) with basement membrane. Methods: Dermal and epidermal layer were separated by immersing in dipase solution for 30 minutes, and the separated layers were treated with 0.05% trypsin for 10 minutes. And then each layer was cultivated to fibroblasts and keratinocytes on a culture medium. Fibroblasts were first penetrated into basement membrane of allogenic dermis facing down, then allogenic dermis was flipped over to face up and keratinocytes were transplanted to allogenic dermis. Results: Observing artificial skin fabricated in vitro, we found following: 1) The artificial skin opened in air for 5 days formed epidermal layer. In dermal layer, fibroblast was distributed evenly among all. 2) The artificial skin opened in air for 30 days formed thicker and thicker, and it formed basement membrane, spinous and granular layers. PAS stain to confirm existence of basement membrane showed positive reaction. 3) Cytokeratin 10 stain to confirm the formation of epidermal layer showed positive reaction. 4) The formation of thick keratin, lamellar body and desmosome similar to human skin were observed in result of an electron micrograph. Conclusion: As a result of research, the structure seen in normal skin such as rete ridge, is found in reproduced artificial skin. This type of artificial skin can be used as a useful model for investigating skin disease and for clinical application also.

Development of a highly effective recombinant protein from human collagen type III Alpha 1 (COL3A1) to enhance human skin cell functionality

  • Young Un Kim;HyunJoon Gi;Eun Kyung Jeong;Seokwon Han;Woo-Young Seo;Young Jun Kim;Sang Bae Lee;KyeongJin Kim
    • BMB Reports
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    • v.57 no.9
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    • pp.424-429
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    • 2024
  • Collagen type III, a member of the fibrillar collagen group, is a major component of the extracellular matrix in various internal organs, the vascular systems, and skin. It is essential to maintain the structural integrity and functionality of these tissues, and plays a significant role in wound healing, often found alongside collagen type I. Despite being the second most abundant collagen in human tissues after type I, its biological functions on various skin properties have not been thoroughly studied. In this study, we have isolated and developed an effective recombinant protein derived from human collagen type III alpha 1 chain (hCOL3A1). Our findings demonstrate that the recombinant proteins hCOL3A1-THR-M1 and M4 stimulate cell proliferation and collagen biosynthesis in human dermal fibroblasts (HDFs), and enhance wound healing. Notably, hCOL3A1-THR-M1 (referred to as HUCOLLATIN3) specifically penetrates both the epidermal and dermal layers in a full-thickness skin model. These results collectively indicate that hCOL3A1-THR-M1 holds promise as a potential biomaterial to prevent skin aging.