• Title/Summary/Keyword: Human dermal fibroblasts

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The Effect of a Long-Term Cyclic Strain on Human Dermal Fibroblasts Cultured in a Bioreactor on Chitosan-Based Scaffolds for the Development of Tissue Engineered Artificial Dermis

  • Lim, Sae-Hwan;Son, Young-Sook;Kim, Chun-Ho;Shin, Heung-Soo;Kim, Jong-Il
    • Macromolecular Research
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    • v.15 no.4
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    • pp.370-378
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    • 2007
  • Mechanical stimulation is known to activate several cellular signal transduction pathways, leading to the induction of signaling molecules and extracellular matrix (ECM) proteins, thereby modulating cellular activities, such as proliferation and survival. In this study, primary human dermal fibroblasts (HDFs) were seeded onto chitosan-based scaffolds, and then cultured for 3 weeks in a bioreactor under a cyclic strain of 1 Hz frequency. Compared to control samples cultured under static conditions, the application of a cyclic strain stimulated the proliferation of HDFs in I week, and by week 3 the thickness of the cell/scaffold composites increased 1.56 fold. Moreover, immunohistochemical staining of the culture media obtained from the cell/scaffold samples subjected to the cyclic strain, revealed increases in the expression and secretion of ECM proteins, such as fibronectin and collagen. These results suggest that the preconditioning of cell/scaffold composites with a cyclic strain may enhance the proliferation of HDFs, and even facilitate integration of the engineered artificial dermal tissue into the host graft site.

ROLE OF FIBROBLASTS IN ORGANOTYPIC CULTURES OF IMMORTALIZED HUMAN ORAL KERATINOCYTES (섬유모세포의 종류에 따른 불멸화된 구강 각화세포의 삼차원적 배양에 관한 연구)

  • Cheong, Jeong-Kwon;Yoon, Kyu-Ho;Kim, Eun-Cheol
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.3
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    • pp.211-220
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    • 2007
  • Objective: In organotypic culture of immortalized human oral keratinocytes (IHOK), the change of the growth and differentiation was investigated according to the fibroblast type and the involvement of mitogen-activated protein (MAP) kinase. Materials & Methods: IHOK was cultured three dimensionally with gingival fibroblast (GF), dermal fibroblast (DF) and immortalized gingival fibroblast (IGF). We characterized biologic properties of three dimensionally reconstructed IHOK by histological, immunohistochemical, and Western blot analysis. We also investigated whether MAP kinase pathway was involved in epithelial-mesenchymal interaction by Western blot analysis. Results: The best condition of three dimensionally cultured IHOK was the dermal equivalent consisting of type I collagen and IGF. IGF increased the expression of more proliferating cell nuclear antigen (PCNA), involucrin than GF and DF in response to co-culture with IHOK. Extracellularly regulated kinase (ERK) pathway was activated in organotypic co-culture with IGF. Conclusion: The organotypic co-culture of IHOK with dermal equivalent consisting of type I collagen and IGF resulted in excellent morphologic and immunohistochemical characteristics and involved ERK pathway. The epithelial-mesenchymal interaction was activated according to the fibroblast type.

Anti-oxidant and Anti-aging Activities of Sericinjam Gland Hydrolysate Extract in Human Dermal Fibroblasts (사람 섬유아세포에서 세리신잠 실샘가수분해물(Sericinjam Gland Hydrolysate)의 항산화 및 항노화 효과)

  • Cheon, Yuri;Hwang, Jung Wook;Lee, Heui Sam;Yun, Seiyoung;Choi, Yong-Soo;Kang, Sangjin
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.39 no.1
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    • pp.9-17
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    • 2013
  • We studied the anti-oxidant and anti-aging activities of Sericinjam Gland Hydrolysate (SJGH) in the human dermal fibroblasts. SJGH effectively defended cell death and ROS generation under high H2O2 in human dermal fibroblasts. Moreover SJGH reduced the expression of SA-${\beta}$-Gal and MMP-1 under low concentration of $H_2O_2$ whereas biosynthesis of procollagen-I was increased. This results demonstrate the anti-oxidant and anti-aging activities of SJGH. SJGH could be a good candidate for anti-aging cosmetics ingredient.

Effect of Saponin with Antioxidant Activity on Matrix Metalloproteinase in Human Dermal Fibroblasts (항산화 효능을 가진 사포닌이 사람섬유아세포에서 기질 금속 단백질 분해효소에 미치는 영향)

  • Park, Hye-Jung;Kim, Moon-Moo;Lee, Dong-Hwan
    • Journal of Life Science
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    • v.21 no.9
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    • pp.1266-1273
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    • 2011
  • Saponin is a main component of ginseng widely known as an oriental traditional medicinal ingredient. A variety of biological effects of saponin has been reported, but its action related to skin regeneration has remained unclear so far. In this study, the effect of saponin on matrix metalloproteinase as well as its antioxidant effect in cell free system was examined in human dermal fibroblasts. First of all, as a result of investigating the effect of saponin on cell viability using MTT assay, it was shown to increase cell viability below 10 ${\mu}g$/ml, but it also showed cytotoxicity above 25 ${\mu}g$/ml. The antioxidant effect of saponin was exerted by inhibition of $H_2O_2$ in addition to reducing power above 1 ${\mu}g$/ml. In particular, saponin showed a protective effect on DNA oxidation. Furthermore, it was observed that saponin activates MMP-2 and increases MMP-1 activity in gelatin and casein zymography analyses, respectively, indicating that saponin could have potential a therapeutic agent for anti-aging and skin regeneration.

Inhibition of Oxidative Damage by Phlorotannins from Ecklonia cava in Normal Human Dermal Fibroblasts

  • Kim, Moon-Moo;Rajapakseb, Niranjan;Kim, Se-Kwon
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.2
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    • pp.126-135
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    • 2006
  • Phlorotannins which is oligomeric polyphenol of phloroglucinol unit were isolated from solvent fractions of methanolic extract of the brown alga, Ecklonia cava (EC). The inhibitory effects of phlorotannins from EC solvent fractions on oxidative stress were examined in human dermal fibroblasts (HDFs) related to wrinkle formation. Among the solvent fractions, phlorotannins from ethyl acetate fraction exerted the highest scavenging effect on DPPH radical, hydroxyl radical and alkyl radical analyzed by electron spin resonance (ESR) spectroscopy. The levels of intracellular reactive oxygen species (ROS) and lipid peroxidation were measured using 2',7'-dichlorofluorescin diacetate (DCFH-DA) and diphenyl-1-pyrenylphosphine (DPPP), respectively. Their levels were significantly decreased in the presence of phlorotannins from ethyl acetate fraction, compared with other fractions obtained from EC extract (P < 0.01). Furthermore, intracellular glutathione (GSH) level was significantly increased in a time dependent manner by the phlorotannins. Therefore, these results suggest that phlorotannins from EC extract could have a therapeutic potential for prevention and treatment of several diseases such as wrinkle formation related to oxidative stress.

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Protective Effects of Antioxidant Active Fractions Derived from the Edible Seaweed Hizikia fusiformis in Oxidatively Stressed Human Dermal Fibroblasts (식용 해조류 톳(Hizikia fusiformis) 유래 항산화 활성분획물의 산화적 손상이 유도된 인간피부섬유아세포 보호 효과)

  • Cui, Yong Ri;Kim, Hyun-Soo;Je, Jun-Geon;Wang, Lei;Oh, Jae-Young;Jia, Liu;Jeon, You-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.52 no.1
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    • pp.35-42
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    • 2019
  • We investigated the protective effects of antioxidant fractions from a 70% ethanolic extract of Hizikia fusiformis in human dermal fibroblasts (HDFs). Powdered H. fusiformis was extracted with 70% ethanol and then partitioned into three fractions according to polarity using n-hexane (HFH), chloroform (HFC), and ethyl acetate (HFEA). Antioxidant activity was observed in HFEA at 0.66 mg/mL based on the half maximal inhibitory concentration ($IC_{50}$) of 1,1-diphenyl-2-picrylhydrazyl (DPPH), and at 0.24 mg/mL based on alkyl radical scavenging. The protective effects of the HFEA antioxidant fraction against 2,2-azobis-(2-amidinopropane) dihydrochloride (AAPH)-damaged HDFs and the expression of Type I procollagen in HDFs were examined. HFEA caused the proliferation of HDFs with and without AAPH treatment and protected against AAPH damage to HDFs in a dose-dependent manner ($50-200{\mu}g/mL$). This implies that the antioxidant properties of the fractions depended on their proliferative and protective effects. The HFEA antioxidant fraction had significant effects and caused the dose-dependent expression of Type I procollagen, an important anti-wrinkle protein, in HDFs. In conclusion, antioxidant substances in H. fusiformis were found in the ethyl acetate fraction, and the resulting HFEA may have cosmetic applications.

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.

Application of a Composite Skin Equivalent using Collagen and Acellular Dermal Matrix as the Scaffold in a Mouse Model of Full-thickness Wound (콜라겐과 무세포진피를 이용한 혼합형 인공피부 개발 및 쥐 모델에서 창상치료 적용)

  • Lee, Dong Hyuck;Youn, Jin Chul;Lee, Jung Hee;Kim, In Seop
    • KSBB Journal
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    • v.29 no.1
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    • pp.42-49
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    • 2014
  • The aim of this study was to develop a composite human skin equivalent for wound healing. Collagen type1 and acellular dermal matrix powder were utilized as the scaffold with dermal fibroblasts and keratinocytes for the development of a composite human skin equivalent. Fibroblast maintained the volume of composite skin equivalent and also induced keratinocytes to attach and proliferate on the surface of composite skin equivalent. The composite human skin equivalent had a structure and curvature similar to those of real skin. Balb-C nu/nu mice were used for the evaluation of full-thickness wound healing effect of the composite human skin equivalent. Graft of composite skin equivalent on full-thickness wound promoted re-epithelialization and granulation tissue formation at 9 days. Given the average wound-healing time (14 days), the wound in the developed composite skin equivalent healed quickly. The overall results indicated that this three-dimensional composite human skin equivalent can be used to effectively enhance wound healing.

Contraction Behavior of Collagen Gel and Fibroblats Activity in Dermal Equivalent Model

  • Yang, Eun-Kyung;Lee, Doo-Hoon;Park, Sue-Nie;Choe, Tae-Boo;Park, Jung-Keug
    • Journal of Microbiology and Biotechnology
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    • v.7 no.4
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    • pp.267-271
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    • 1997
  • We developed a dermal equivalent (DE) which was engineered using human dermal fibroblasts and a matrix of collagen gel. The in vitro construction of the DE was accomplished by casting a porcine collagen type I solution plus concentrated medium with isolated and cultured fibroblasts. These constructs were attached to culture dishes or left floating in culture medium. Contraction of attached gels results in decreased gel thickness without a change in gel diameter, and contraction of floating gels results in decreased gel thickness and diameter. After contraction, there was no increase in cell number in floating gels, but cells in attached gels began to increase after about 4 days of the lag phase in cell growth curve. At this lag phase, addition of fibroblast growth factor (FGF) at a concentration of $0.1{\mu}$/ml promoted cell proliferation in the attached collagen gels, but no effect in floating gels. These results indicate that the method of contraction had an influence on the extracellular matrix (ECM) organization, and this influenced not only cell growth but also fibroblast responsiveness to FGF. This suggests that attached collagen gel is more suitable as a dermal equivalent than the floating gel. And the final contracted area of attached gel is much larger than that of the floating gel since floating gel is contracted in all directions but attached gel is contracted only vertically.

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Stimulation of the Extracellular Matrix Production in Dermal Fibroblasts by Areca catechu Extract (진피섬유모세포에서 대복피추출물의 세포외기질 합성 촉진 효과)

  • Lee, Min-Ho;Kim, Hyung-Jin;Jung, Hyun-Ah;Lee, Young-Keun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.4
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    • pp.1857-1862
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    • 2013
  • Dermal fibroblasts produce the many components of the extracellular matrix (ECM) that are needed to maintain connective tissue integrity and repair tissue injuries. This study investigated the effects of Areca catechu extract (ACE) on dermal fibroblast cell activation. Cultured human dermal fibroblasts were treated with ACE, and then ECM production was determined by ELISA, Western blot and RT-PCR. ACE significantly accelerated the production of type 1 collagen, fibronectin, and transforming growth factor (TGF)-${\beta}1$ by ELISA and type 1 collagen by Western blot assay. ACE also increased the gene expression of COL1A1, TGF-${\beta}1$, keratinocyte growth factor (KGF) and insulin growth factor (IGF)-1. These results suggest that ACE has the potential to stimulate ECM production and that it might be suitable for maintaining skin texture.