• Radiation Oncology Journal
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• v.17 no.2
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• pp.158-165
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• 1999

Purpose : To investigate the percentage of colonies wi1h16or more cells distribution of human skin fibroblast according to in vitro aging, and to evaluate the relationship between percentage of colonies with 10 or more cells and in vivo donor age in human skin fibroblast culture. Material and Method : C1, C2, C3a, and C3b human skin fibroblast samples from three breast cancer patients were used as subjects. The C1, C2, and C3a donor were 44, 54, and 55 years old, respectively. C3a and C3b cells were isolated from the same person. Single cell suspension of skin fibroblasts was prepared with primary explant technique. One hundred cells are plated into 100m1 tissue culture flask and cultured for two weeks. The colony size was defined as colonies with 16 or more cells. The cultured cell was stained with crystal violet, and number of cells in each colony was determined with stereo microscope at $\times$10 magnification. Passage number of C1, C2, C3a and C3b skin fibroblast were 12th, 17th, and 14th, respectively. Results : Percentage of colonies with 16 or more cells of skin fibroblast samples decreased with increasing in vitro passage number. In contrast, cumulative population doublings of skin fibroblast sample increased with increasing in vitro passage number. Percentage of colonies with 16 or more cells also decreased with increasing population doublings in human skin fibroblast culture. There was strong correlation with percentage of colonies with 16 or more cells and population doublings En C3a skin fibroblast sampie. At the same point of population doublings, the percentage of colonies with 16 or more cells of the young C1 donor was higher level than the old C3a donor. Conclusion : The population doublings increased with increasing in vitro passage number but percentage of colonies with 16 or more cells decreased. The results of this study imply that percentage of colonies with 16 or more cell is useful as a indicator of in vitro human skin fibroblast aging and may estimate the in vivo donor age.

• Journal of the Korean Applied Science and Technology
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• v.36 no.2
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• pp.394-406
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• 2019

The cellular senescence may be due to damage by the reactive oxygen species (ROS). This study has compared the antioxidant activity in the human cell lines of various origins, including 10S and 50S-derived normal skin fibroblasts, and 10S bone marrow, dental tissue and adipose-derived adult stem cells. After being exposed to $H_2O_2$, half inhibitory concentration ($IC_{50}$) values by cytotoxicity assay was significantly (P<0.05) lower in 50S-derived skin fibroblasts, than in 10S-derived skin fibroblasts and various adult stem cell lines. The cell population doubling time (PDT) and the cell frequency with high senescence associated-${\beta}$-galactose activity were remarkably increased in 50S-derived fibroblasts exposed to 50 ppm $H_2O_2$ for 7 days, than those of 10S-derived fibroblasts and various adult stem cell lines. Further, the expression level of antioxidant-related genes, glutathione peroxidase (GPX) and catalase (CAT), was investigated in 10S and 50S-derived skin fibroblasts, and 10S-derived various adult stem cells by reverse transcription polymerase chain reaction (RT-PCR). The expression level of GPX was higher in most of cell lines, compared to CAT, and a significantly (P<0.05) higher expression level of GPX was observed in 10S-derived skin fibroblasts and adult stem cell lines, compared to 50S-derived skin fibroblasts. We concluded that old-aged skin fibroblasts seemed to be less resistant against ROS than young-aged skin fibroblasts and adult stem cells.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.40 no.3
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• pp.279-287
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• 2014

Estrogen deficiency results in a reduction of skin quality and function in postmenopausal women. Over the past decade, many studies have supported that estrogen provides anti-aging effects as a result of the ability of estrogen to prevent skin collagen decline, restore skin elasticity, and increase skin hydration in postmenopausal women skin. Due to their structural similarity with estrogen, isoflavones have been called phytoestrogens. Photoprotective effects of isoflavones are well established while their estrogenic-like activities are not fully understood in human skin. In this study, we investigated whether daidzein, an effective isoflavone, has phytoestrogenic activity and induces transcriptional change of extracellular matrix components in dermal fibroblasts. We examined the luciferase activity of daidzein and ${\beta}$-estradiol using transiently transfected NIH3T3-ERE cells. The estrogenic receptor-dependent transcriptional activity was increased in a dose-dependent manner when treated with daidzein, with a maximum of 2.5-fold induction at $10{\mu}g/mL$ of daidzein compared with non-treated control. In addition, daidzein significantly in creased the expressions of collagen type I, collagen type IV, elastin, and fibrillin-1 in human dermal fibroblasts. By comparing with the effects of ${\beta}$-estradiol through out all the experiments, we confirmed that daidzein had estrogenic activity and function in fibroblasts. These results suggest that daidzein-based application, having both photoprotective and phytoestrogenic effects, may be a powerful approach for skin anti-aging of postmenopausal women.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.38 no.3
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• pp.219-224
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• 2012

European Union prohibited the marketing of cosmetic products containing constituents that have been examined through animal experiments. Thus, non-animal test models are needed to replace animal experiments. The reconstructed skin models are important as a test system for cosmetic, pharmaceutical, and medical device safety testing. In the present study, we tried to develop an optimal skin equivalent model containing basement membrane and epidermis. For this purpose, we used mesenchymal stem cells (MSCs) and/or preadipocytes as well as fibroblasts as the dermal matrix cells. The formation of basement membrane and epidermis was verified by immunohistochemical stains. Among various models, the epidermis was thickest when MSCs were used in the dermal matrix. Furthermore, PCNA and involucrin distribution showed that dermal matrix with MSCs resembled human skin. Therefore, skin equivalents with MSCs could be developed as a non-animal test model to replace animal experiments.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.4 s.48
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• pp.499-502
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• 2004

DPPH radical-generating system was used to evaluate the antioxidant properties of the Rosaceae. The inhibitory effects of ethanolic extracts from Rosaceae plants were investigated on melanin biosynthesis which is closely related to hyperpigmentation. Of the Rosaceae extracts, Prunus sargentii, Rubus coreanus, Chaenomeles sinensis, Photinia glabra and Pyrus pyrifolia showed a potent inhibition of tyrosinase, the enzyme which converts 3-(3,4-dihydroxyphenyl) alanine (dopa) to dopachrome in the melanin biosynthetic process. Furthermore, MMT assay was used to check the cytotoxicity of extracts on the human foreskin fibroblast cell line, Hs68. Among the Rosaceae, bark of Prunus sargentii, bark wood of Photinia glabra and all parts or Chaenomeles sinensis showed more than $50\%$ inhibition of mushroom tyrosinase activity at 100 ug/mL and more than $80\%$ of strong DPPH radical-scavenging activity at 10 ug/mL. In audition to, they had no cytotoxic activity on Hs68. These results suggest that these extracts might be except a controler in pigmentation.

• 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.

• Journal of Ginseng Research
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• v.31 no.2
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• pp.86-92
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• 2007

This study was carried out to develop health & functional food by using Korean red ginseng for prevention of skin wrinkles. Effects of Korean red ginseng on the collagen biosynthesis and inhibition of matrix metalloproteinase-I (MMP-1) activity in human dermal fibroblast were investigated. Crude saponin contents of Korean red ginseng water extract (WE), Korean red ginseng ethanol extracts (EE) and Korean Red ginseng purified extracts (PE) were 72 mg/g, 107 mg/g and 220 mg/g, respectively. We incubated human fibroblast cell with Korean red ginseng component by addition of l ${\mu}g/ml$, 5 ${\mu}g/ml$, 10 ${\mu}g/ml$. Amount of collagen biosynthesis was 1.86 ng/ml in control sample and 2.85 ng/ml, 2.05 ng/ml and 2.58 ng/ml in retinoic acid, EE and PE respectively. Furthermore, $ginsenoside-Rg_1$ and $ginsenoside-Rb_1$ were shown 2.01 ng/ml and 3.07 ng/ml. MMP-l activities of EE, PE, $ginsenoside-Rg_1$ and $ginsenoside-Rb_1$ were decreased to 92%, 94%, 91% and 78% respectively as compared with control. Cell proliferation were showed 84-96% in the Korean red ginseng components. The antioxidative SOD activities of the Korean red ginseng components were showed 28-69%, however it was lower than that of Vitamin C. From this results, we conclude that Korean red ginseng have a anti-wrinkle effect and $ginsenoside-Rb_1$ may be considered as a more effective component.

• Journal of Korean society of Dental Hygiene
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• v.16 no.3
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• pp.471-477
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• 2016

Objectives: Fluoride is widely used in the prevention and control of dental caries. The purpose of this study is to examine the biological effects of Sodium fluoride on the proliferation of oral normal cell in vitro(MDPC-23, HaCaT, HGF-1 cells). Methods: The proliferation of normal cells and the cyto-skeletal change of normal cells were assessed by WST-1 assay and F-actin stain assay. The statistical significances of the resulting data were analyzed using SPSS(Window 12.0). Results: The sodium fluoride(0-12 mM) treatment decreased the cell viability in a dose and time dependent manner: HaCaT(6 h): $100{\pm}0$, $98{\pm}0.39$, $82{\pm}2.68$, $75{\pm}0.83$, $69{\pm}1$, $67{\pm}1.42%$(p<0.005); HaCaT(24 h): $100{\pm}0$, $98{\pm}1.85$, $54{\pm}0.64$, $43{\pm}0.4$, $38{\pm}0.32$, $36{\pm}0.13%$(p<0.006), MDPC-23(6 h): $100{\pm}0$, $93{\pm}1.48$, $85{\pm}0.28$, $82{\pm}1.58$, $79{\pm}1.48$, $76{\pm}1.93%$(p<0.009); MDPC-23(24 h): $100{\pm}0$, $91{\pm}1.26$, $58{\pm}0.65$, $49{\pm}1$, $44{\pm}0.74$, $2{\pm}0.05%$(p<0.005), HGF-1(6 h): $100{\pm}0$, $97{\pm}2.93$, $89{\pm}5$, $71{\pm}5.42$, $58{\pm}4.82$, $43{\pm}3.47%$(p<0.009); HGF-1(24 h): $100{\pm}0$, $97{\pm}2.05$, $73{\pm}1.73$, $22{\pm}1.61$, $14{\pm}1.73$, $7{\pm}0.85%$(p<0.005). Thus, changes in cell morphology and disruption of filamentous(F)-actin organization were observed in higher concentration. Conclusions: These results suggest that higher concentrations of fluoride lead to a reduce the number of cells and morphology change of normal cell.