인체피부섬유아세포에서 애기달맞이꽃 메탄올추출물의 항 주름 효능

김수은 ( Su Eun Kim ),이채명 ( Chae Myoung Lee ),김영철 ( Young Chul Kim ) 대한미용학회(구 대한미용과학회) 2016 대한미용학회지 Vol.12 No.3

The objective of this study was to evaluate the anti-wrinkle efficacy of Oenothera laciniata methanol extract (OLME) in vitro using human dermal fibroblasts. Specifically, we tested the type-I procollagen synthetic ability of OLME at concentrations of 6.25, 12.5, 25, and 50 μg/mL and noted significantly significant (p<0.05) increases versus the control of 38.7, 74.3, 111.4, and 113.0%, respectively. Using RT-PCR, the expression of MMP-1 mRNA in human dermal fibroblasts treated with 6.25, 12.5, 25, and 50 μg/mL OLME was shown to have decreased when compared to the control by 67.7, 71.7, 75.6, and 84.3%, respectively (p<0.05). When measuring MMP-2 activity in human dermal fibroblasts treated with 6.25, 12.5, 25, and 50 μg/mL OLME using zymography, it was shown to have decreased versus the control by 27.6, 37.1, 76.3, and 93.9%, respectively (p<0.05). OLME was less cytotoxic to human dermal fibroblasts than ascorbic acid. From these results, we found that OLME is highly efficacious in its ability to promote collagen synthesis and inhibit its degradation in human dermal fibroblasts, similar or even superior to those of ascorbic acid, a well-known anti-skin aging agent. Therefore, OLME could be used as a functional cosmetic material for the prevention or alleviation of skin wrinkles induced by ultraviolet rays.

Uniaxial cyclic strain stimulates cell proliferation and secretion of interleukin-6 and vascular endothelial growth factor of human dermal fibroblasts seeded on chitosan scaffolds.

Park, Ki-Sook,Lee, Eun-Gyu,Son, Youngsook John Wiley Sons 2014 Journal of biomedical materials research. Part A Vol.102 No.7

<P>Human dermal fibroblasts were inoculated into chitosan sponge scaffolds coated with type I collagen and it might be developed as a dermal substitute and/or dressing material. The application of 14% uniaxial cyclic strain to the cellular scaffolds affected the characteristics of the seeded human dermal fibroblasts. Cyclic strain enhanced cellular proliferation, the activity of metalloproteinase-2, and the expression of extracellular matrix proteins such as fibronectin. Moreover, cyclic strain increased the expression of vascular endothelial growth factor (VEGF) and interleukin (IL)-6, which are critical to wound healing. Even under static culture (strain, 0%) following 14% cyclic strain, the expression of VEGF and IL-6, which had increased under 14% strain, was amplified or maintained for at least 3 days. Uniaxial cyclic strain may enhance the wound-healing potential of human dermal fibroblasts seeded on chitosan scaffolds through the changes in the cellular characteristics of the fibroblasts when the cellular scaffold is transplanted into skin wounds, especially chronic wounds such as diabetic wounds.</P>

인체섬유아세포를 이용한 헐리웃과 홍로센 자두 과피 열수추출물의 콜라겐 합성능 및 분해능 평가

김상남 ( Sang Nam Kim ),김영철 ( Young Chul Kim ) 한국미용학회 2013 한국미용학회지 Vol.19 No.2

The purpose of this study was to evaluate collagen synthetic and disintegrating efficacy of water-soluble extracts of hollywood (HW) and hongrosen`s (HR) plum peel using human dermal fibroblasts. The maximum permissible level for HW and HR to human dermal fibroblasts was over 200 μg/mL. In the experiment of type-I procollagen synthetic ability, at the concentrations of 12.5, 25, 50 and 100 μg/mL, HW increased it by 26.3 (p<0.05), 43.8 (p<0.01), 52.0 (p<0.001) and 52.0% (p<0.001), and HR significantly (p<0.001) increased it by 53.6, 54.2, 54.7 and 56.9%, respectively, compared to the control group. In the experiment of matrix metalloproteinas -2 (MMP-2) activity, at the concentrations of 12.5, 25, 50 and 100 μg/mL, HW significantly (p<0.001) decreased it by 29.6, 32.9, 36.1 and 38.0%, and HR significantly (p<0.001) decreased it by 28.5, 30.1, 31.5 and 34.3%, respectively, compared to the control group. These results showed that HW and HR had very low toxicity to human dermal fibroblasts, and had high efficacy on the stimulation of collagen synthesis and inhibition of collagen degradation in human dermal fibroblasts. Taken the results in consideration, HW and HR could be used as functional cosmetic materials for the prevention or alleviation of skin aging induced by ultraviolet rays.

Antioxidant effect of 23-hydroxytormentic acid on human dermal fibroblasts

( Young Ah Cho ),( Ji Su Lee ),( Sung Min Kim ),( Ji Youn Hong ),( Jin Hee Kim ),( Joo Ran Hong ),( Hye In Cheon ),( Hae Jeong Youn ),( Yang Won Lee ),( Yong Beom Choe ),( Kyu Joong Ahn ) 대한피부과학회 2020 대한피부과학회 학술발표대회집 Vol.72 No.1

Background: Reactive oxygen species (ROS) are generated by Ultraviolet A (UVA) in the dermal layer of the skin. ROS causes inflammatory change of dermal tissue, photoaging, and alterations of gene expression. For this reason, how to protect skin from UVA has been continuously investigated. Objectives: This study aimed to investigate the effect of 23-Hydroxytormentic acid to UVA-irradiated human dermal fibroblasts in vitro. Methods: The antioxidant effect of 23-hydroxytormentic acid (23-HTA) was evaluated by using 2,7-dichlorofluorescin diacetate and 3-(4,5-dimethylthiazol- 2-yl)-2,5,diphenyltertazolium bromide. By using quantitative real-time polymerase chain reaction, mRNA levels of antioxidant molecule were investigated. Results: 23-HTA was associated with antioxidant effect on human dermal fibroblast from UVA by regulating antioxidant-related gene expression and ROS scavenging. In addition, the SA-β-gal analysis and results of mRNA levels of type I procollagen and matrix mellaoproteinases demonstrated that 23-HTA attenuates UVA-induced oxidative stress by regulating the extracellular matrix proteins gene expression and these results were statistically significant. Conclusion: This study suggests that 23-HTA has protecting effect against UVA-induced oxidation stress on human dermal fibroblast. In order to clarify the antioxidant effect of 23-HTA, in vivo study is needed in the future.

Human Dermal Stem/Progenitor Cell-Derived Conditioned Medium Improves Senescent Human Dermal Fibroblasts

Jung, Ji-Yong,Shim, Joong Hyun,Choi, Hyun,Lee, Tae Ryong,Shin, Dong Wook MDPI 2015 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.16 No.8

<P>Adult skin stem cells are recognized as potential therapeutics to rejuvenate aged skin. We previously demonstrated that human dermal stem/progenitor cells (hDSPCs) with multipotent capacity could be enriched from human dermal fibroblasts using collagen type IV. However, the effects of hDSPCs on cellular senescence remain to be elucidated. In the present study, we investigated whether conditioned medium (CM) collected from hDSPC cultures (hDSPC-CM) exhibits beneficial effects on senescent fibroblasts. We found that hDSPC-CM promoted proliferation and decreased the expression level of senescence-associated β-galactosidase in senescent fibroblasts. In addition, p53 phosphorylation and p21 expression were significantly reduced in senescent fibroblasts treated with hDSPC-CM. hDSPC-CM restored the expression levels of collagen type I, collagen type III, and tissue inhibitor of metalloproteinase, and antagonized the increase of matrix metalloproteinase 1 expression. Finally, we demonstrated that hDSPC-CM significantly reduced reactive oxygen species levels by specifically up-regulating the expression level of superoxide dismutase 2. Taken together, these data suggest that hDSPC-CM can be applied as a potential therapeutic agent for improving human aged skin.</P>

히알루론산이 섬유아세포의 증식 및 상처 치유에 미치는 영향

한승규,김정배,박현,이병일,김우경 대한성형외과학회 2002 Archives of Plastic Surgery Vol.29 No.3

It is not determined yet whether hyaluronic acid up- regulates or down-regulates wound healing. This study was designed to define the effect of hyaluronic acid on proliferation of human dermal fibroblasts in vitro and on skin wound closing in vivo.Fibroblasts were isolated from the dermis of adults and cultivated in the presence of either one of 6 concentrations of hyaluronic acid(0, 0.1, 0.2, 0.5, 1.0, 2.0mg/ml). The fibroblasts were seeded at 2.0×10⁴cells/well in Dulbecco's Modified Eagle's Medium/Ham's F-12 nutrient including 10% fetal bovine serum with either one of 6 different concentrations of hyaluronic acid in 24-well plates. The cells were incubated for 6 days. All concentrations of hyaluronic acid stimulated the proliferation of fibroblasts. The best proliferation was seen at 0.2mg/ml of hyaluronic acid concentration(p=0.01). For in vivo study, 10 white rats were used. A 5mm round punch was employed to excise skin and subcutaneous tissue at eight sites on the back. After creating 8 open wounds, 8 concentrations(0, 0.1, 0.2, 0.5, 1.0, 2.0, 5.0, 10.0mg/ml) of hyaluronic acid were applied. The degrees of wound closing were compared the 6th day under light microscope. Low concentration of hyaluronic acid(0-2.0mg/ml) stimulated the wound closing. However, high concentration of hyaluronic acid(5.0-10.0 mg/ml) delayed the wound closing. The best wound closing was seen at 0.5mg/ml of hyaluronic acid(p=0.032). These results demonstrated that hyaluronic acid influenced human dermal fibroblast proliferation and the skin wound closing in rats, and its concentration was critically important factor.

Peptide Hydrolysates from Astragalus membranaceus Bunge Inhibit the Expression of Matrix Metalloproteinases in Human Dermal Fibroblasts

Sun Ki Park(박선기),Pham Van Hien,Hoang Van Luong,Shao-wei Yan,Sang Yo Byun(변상요) 한국생물공학회 2014 KSBB Journal Vol.29 No.5

Inhibition effects of peptide hydrolysates from Astragalus membranaceus Bunge. on the expression of the matrix metalloproteinases (MMPs) in human dermal fibroblasts were evaluated in vitro. Crude peptides were obtained by the hydrolysis of proteins extracted from A. membranaceus. Peptides were purified partially by the basis on the molecular weight using 40% polyacrylamide gel electrophoresis before treatment with human dermal fibroblasts. Basis on the doseeffect experiments, expressions of MMPs including MMP-1, MMP-3, MMP-8, MMP-13 in human dermal fibroblasts were evaluated. Expressions of MMP-1, MMP-3, MMP-8 and MMP-13 were reduced in 43%, 5%, 22% and 57% respectively. The mass spectrometric analysis of partially purified peptides from A. membranaceus, which strongly inhibit expressions of MMPs, indicated that the peptides were composed of molecules below 1500 Da.

Peptide Hydrolysates from Astragalus membranaceus Bunge Inhibit the Expression of Matrix Metalloproteinases in Human Dermal Fibroblasts

Park, Sun Ki,Van Hien, Pham,Van Luong, Hoang,Yan, Shao-Wei,Byun, Sang Yo The Korean Society for Biotechnology and Bioengine 2014 KSBB Journal Vol.4 No.1

Inhibition effects of peptide hydrolysates from Astragalus membranaceus Bunge. on the expression of the matrix metalloproteinases (MMPs) in human dermal fibroblasts were evaluated in vitro. Crude peptides were obtained by the hydrolysis of proteins extracted from A. membranaceus. Peptides were purified partially by the basis on the molecular weight using 40% polyacrylamide gel electrophoresis before treatment with human dermal fibroblasts. Basis on the doseeffect experiments, expressions of MMPs including MMP-1, MMP-3, MMP-8, MMP-13 in human dermal fibroblasts were evaluated. Expressions of MMP-1, MMP-3, MMP-8 and MMP-13 were reduced in 43%, 5%, 22% and 57% respectively. The mass spectrometric analysis of partially purified peptides from A. membranaceus, which strongly inhibit expressions of MMPs, indicated that the peptides were composed of molecules below 1500 Da.

The Reconstruction of Skin Equivalents with Hair Follicle Dermal Sheath Cells

( Hyun Joo Cho ),( Hye Ryung Choi ),( In Hwa Bae ),( Hyun Joo Chung ),( Dong Seok Kim ),( Sun Bang Kwon ),( Jung Im Na ),( Sang Woong Youn ),( Kyoung Chan Park ) 한국조직공학과 재생의학회 2004 조직공학과 재생의학 Vol.1 No.2

The skin consists of two main layers, the epidermis and dermis, which are separated by the basement membrane. Epidermal-dermal communication is important for skin homeostasis, and dermal factors play a key role in regulating the growth and differentiation of the epidermis. Several studies have demonstrated that hair follicle dermal sheath cells (DSCs) synthesize more soluble factors than dermal fibroblasts. The aim of this study is to investigate the effects of DSCs upon the evolution of skin equivalents (SEs). In this study, human keratinocytes were seeded on two different dermal substitutes. In addition, the effects of ascorbic acid were evaluated. Histologically, thick epidermis was observed in DSC-derived models. Immunohistochemical staining showed similar pattern of involucrin in both DSC- and fibroblast-derived models, but filaggrin was expressed much stronger in DSC models. The number of PCNA positive cells was lower in the DSC model than in the fibroblast model especially in the presence of ascorbic acid. However, integrin a6 was strongly expressed in DSC models and in the presence of ascorbic acid supplementation. These findings suggest that DSCs secrete or synthesize factors, which affect the epidermalization of the skin, and that ascorbic acid is an important supplement in the reconstruction of human epidermis. In addition, transmission electron microscopy of SEs was performed and a well-developed intercellular junction was observed in the DSC models. Our results suggest that dermal compartment-related epidermis modifications are important in the processes of differentiation and proliferation in SEs.

홍삼 생약 복합물(KTNG0345)이 섬유아세포의 Procollagen 생합성과 Matrix metalloproteinase-1(MMP-1) 활성에 미치는 영향

소승호(Seung-Ho So),이성계(Seong-Kye Lee),황의일(Eui-Il Hwang),구본석(Bon-Suk Koo),한경호(Gyeong-Ho Han),김나미(Na-Mi Kim) 고려인삼학회 2007 Journal of Ginseng Research Vol.31 No.3

본 실험은 홍삼, 사상자, 산수유를 이용하여 주름 예방 효과가 있는 건강기능 식품을 개발하기 위한 기초자료로 활용하기 위하여 개별추출물과 이들의 혼합물 WM1, WM3(KTNG0345)에 대하여 사람의 섬유아세포를 이용한 세포증식, 콜라겐 생합성 정도, MMP-1 효소활성, 항산화 활성을 조사하였다. 사용된 시료는 1~10 ㎍/ml의 농도로 첨가하여 세포를 배양했을 때 83~100%의 세포증식율을 나타내어 세포 독성을 나타내지 않았으며, 콜라겐 생합성 정도는 대조군이 474.8 ng/ml이었고 retinoic acid는 568.3 ng/ml이었으며, 5 ㎍/ml의 농도를 기준으로 할 때 홍삼은 533.9 ng/ml, 사상자는 539.3 ng/ml, 산수유는 514.1 ng/ml이었다. 홍삼과 생약을 68 : 232의 비율로 혼합한 WM1은 474.6 ng/ml이었고 홍삼과 생약을 136 : 164의 비율로 혼합한 WM3는 561.4 ng/ml로 증가하였다. MMP-1활성은 10 ㎍/ml의 농도에서 홍삼(ER)은 31.9 ng/ml, 사상자(WT)는 32.85 ng/ml, 산수유(WC)는 32.0 ng/ml, WM1은 31.3 ng/ml이었고 WM3(KTNG0345)는 28.85 ng/ml으로 대조군의 88% 수준으로 감소하였다. SOD활성에서는 1000 ㎍/ml의 농도로 처리하였을 때 65-97% 항산화활성을 나타내었다. 이들의 결과를 종합해 보면 홍삼과 사상자, 산수유 및 이들 혼합물은 human dermal fibroblast에 대하여 세포 독성을 나타내지 않으면서 콜라겐 생합성을 촉진하고 MMP-1의 활성을 억제하였으며 홍삼과 생약 엑기스를 136 : 164의 비율로 혼합했을 때 그 효과가 증진되었다. WM3(KTNG0345)는 지표성분으로서 ginsenoside-Rb1, torilin, loganin을 각각 10.85 ㎎/g, 0.128 ㎎/g, 3.92 ㎎/g 함유하였다. Skin wrinkles are associated with collagen synthesis and matrix metalloproteinase-I (MMP-1) activity. This study was carried out to select optimum ratio of 3 herbs in skin health food for anti-wrinkle. Human dermal fibroblast cell was incubated with experimental samples, which were Korean red ginseng ethanol extracts (ER), Torilis fructus water extracts (WT), Corni fructus water extracts (WC) and their mixtures (WM1, WM3). And then we determined effects on collagen biosynthesis, MMP-1 activity and SOD activity in human dermal fibroblast cell. In control group, collagen biosynthesis was amounted at 474.8 ng/ml and 533.9 ng/ml, 539.3 ng/ml, 514.1 ng/ml in ER, WT and WC respectively. Furthermore, WM3 (KTNG0345) was increased to 561.45 ng/ml. MMP-1 activity of ER, WT, WC, WM1 were determined to 31.9 ng/ml, 32.85 ng/ml, 32.0 ng/ml, 31.3 ng/ml and WM3 (KTNG0345) was decreased to 28.85 ng/ml. In addition, the experimental samples showed a antioxidative activities. From this results, we conclude that Korean red ginseng ethanol extracts, Torilis fructus water extracts, Corni fructus water extracts and their mixtures have a anti-wrinkle effect and WM3 (KTNG0345) may be regarded as an optimum composition for synergic effect producing. The standardized components of KTNG0345, ginsenoside-Rb₁, torilin and loganin were identified at 10.85 ㎎/g, 0.128 ㎎/g and 3.92 ㎎/g respectively.