• Journal of the Society of Cosmetic Scientists of Korea
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• v.40 no.4
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• pp.413-421
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• 2014

Corticotropin-releasing factor (CRF) is involved in the stress response and there is increasing evidence that stress influences skin disease such as hair loss. In cultured human hair follicles, CRF inhibits hair shaft elongation, induces premature regression and promotes the apoptosis of hair matrix keratinocytes. We investigated whether CRF influences the dermal papilla cells (DPC) that play pivotal roles in hair growth and cycling. Human DPCs were treated with CRF, adrenocorticotropic hormone (ACTH) and cortisol, key stress hormones along the hypothalamic-pituitary -adrenal (HPA) axis for 1-24 h. Interestingly, CRF modulated the expression of cytokines related to hair growth (KGF, Wnt5a, $TGF{\beta}-2$, Nexin) and increased cAMP production in cultured DPCs. CRF receptors were down-regulated by negative feedback systems. Pretreatment of CRF receptor antagonists or protein kinase A (PKA) inhibitor prevented the CRF-induced modulation. Since the CRF induces proopiomelanocortin (POMC) expression through the cAMP/PKA pathway, we analyzed POMC mRNA. CRF stimulated POMC expression in cultured human DPCs, yet we were unable to detect ACTH levels by western blot. These results indicate that CRF operates within DPCs through CRF receptors along the classical CRF signaling pathway and CRF receptor antagonists could serve as potential therapeutic and cosmetic agents for stress-induced hair loss.

• Journal of the Korean Wood Science and Technology
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• v.48 no.2
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• pp.166-180
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• 2020

Rhododendron schlippenbachii have been used as a medicine because of their various biological activities. In this study, R. schlippenbachii ethanol extract was evaluated for the treatment of vitiligo. The R. schlippenbachii ethanol extract did not show any cell cytotoxicity. The effect on mushroom tyrosinase and cellular tyrosinase activities were further assessed. In addition, the determination of melanin content in melanocytes was measured using both the B16 melanoma cells and C57BL/6J Ler-vit/vit mice. Finally, the existence of quercetin in R. schlippenbachii was confirmed by qualitative analysis using HPLC. The results clearly demonstrated the R. schlippenbachii extract enhanced melanogenesis and also increased tyrosinase activity in cultured melanoma cells and C57BL/6J Ler-vit/vit mice. In addition, treatment with R. schlippenbachii extract led to a higher content of melanin and eumelanin in C57BL/6J Ler-vit/vit mice hair than in control (untreated) mice, which demonstrated the therapeutic effect of hair-graying associated with vitiligo. Finally, we confirmed a notable increase in melanocytes in the skin of C57BL/6J Ler-vit/vit mice treated with R. schlippenbachii extract compared with the control. Extracts of R. schlippenbachii was shown to be potent tyrosinase and melanin synthesis activator in B16 melanoma cells. The R. schlippenbachii extract have significantly higher melanin content than the untreated control in C57BL/6J Ler-vit/vit mice hair. The results suggest that R. schlippenbachii extract might be considered as an alternative treatment for improvement of vitiligo.

• Restorative Dentistry and Endodontics
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• v.24 no.3
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• pp.437-446
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• 1999

Porphyromonas endodontalis(P. endodontalis) is one of the important causative bacteria of pulpal and periapical disease. P. endodontalis has lipopolysaccharide(LPS) and it plays a major role in stimulating the synthesis and release of cytokines from immune cells and prostaglandin $E_2$ from host cells. The purpose of this study is to prepare LPS from P. endodontalis and to evaluate the effect of LPS on membrane permeability of fibroblast. P. endodontalis ATCC 35406 was cultured in anaerobic condition, and LPS was extracted. LPS was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Human periodontal ligament cell, colon fibroblast(CCD-18Co, KCLB 21459) and skin fibroblast(Detroit 551, KCLB 10110) were perfused with 0.01% P. endodontalis LPS solution, high concentration of $K^+$ solution and $Ca^{2+}$-free solution, $Ca^{2+}$ concentration ratio was measured by microfluorometry. 1. Intracellular $Ca^{2+}$ concentration was not changed in human periodontal fibroblast and skin fibroblast(Detroit 551) stimulated by P. endodontalis LPS. 2. Intracellular $Ca^{2+}$ concentration was increased in colon fibroblast(CCD-18Co) stimulated by P. endodontalis LPS. 3. Colon fibroblast(CCD-18Co) has voltage dependent $Ca^{2+}$ channel activated by high concentration of $K^+$ solution. 4. P. endodontalis LPS has no effect on the increase of intracellular $Ca^{2+}$ concentration during perfusion of $Ca^{2+}$-free solution.

• Archives of Pharmacal Research
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• v.28 no.2
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• pp.195-202
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• 2005

Free radicals and reactive oxygen species (ROS) caused by UV exposure or other environmental factors are critical players in cellular damage and aging. In order to develop a new antiphotoaging agent, this work focused on the antioxidant effects of the extract of tinged autumnal leaves of Acer palmatum. One compound was isolated from an ethyl acetate soluble fraction of the A. palmatum extract using silica gel column chromatography. The chemical structure was identified as apigenin-8-C-beta-D-glucopyranoside, more commonly known as vitexin, by spectral analysis including LC-MS, FT-IR, UV, $^{1}H-$, and $^{13}C-NMR$. The biological activities of vitexin were investigated for the potential application of its anti-aging effects in the cosmetic field. Vitexin inhibited superoxide radicals by about $70\%$ at a concentration of $100\;{\mu}g/mL$ and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals by about $60\%$ at a concentration of $100\;{\mu}g/mL$. Intracellular ROS scavenging activity was indicated by increases in dichlorofluorescein (DCF) fluorescence upon exposure to UVB $20\;mJ/cm^2$ in cultured human dermal fibroblasts (HDFs) after the treatment of vitexin. The results show that oxidation of 5-(6-)chloromethyl-2',7'-dichlo-rodihydrofluorescein diacetate ($CM-H_{2}DCFDA$) is inhibited by vitexin effectively and that vitexin has a potent free radical scavenging activity in UVB-irradiated HDFs. In ROS imaging using a confocal microscope we visualized DCF fluorescence in HDFs directly. In conclusion, our findings suggest that vitexin can be effectively used for the prevention of UV-induced adverse skin reactions such as free radical production and skin cell damage.

• Journal of the Korean Society for Precision Engineering
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• v.32 no.8
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• pp.755-760
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• 2015

For proper wound healing, dermal contraction and remodeling are critical; during the natural healing process, differentiated fibroblasts called "myofibroblasts" typically undertake these functions. For severe wounds, however, a critical mass of dermal matrix and fibroblasts are lost, making self-regeneration impossible. To overcome this impairment, synthetic wound patches with embedded functional cells can be used to promote healing. In this study, we developed a polydioxanone (PDO)-based cell-embedded sheet on which dermal fibroblasts were cultured and induced for differentiation into myofibroblasts, whereby the following combinatorial physicochemical stimuli were also applied: aligned topology, electric field (EF), and growth factor. The results show that both the aligned topology and EF synergistically enhanced the expression of alpha smooth-muscle actin (${\alpha}$-SMA), a key myofibroblast marker. Our proof-of-concept (POC) experiments demonstrated the potential applicability of a myofibroblast-embedded PDO sheet as a wound patch.

• Natural Product Sciences
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• v.17 no.1
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• pp.26-32
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• 2011

Novel tyrosinase inhibitors are important for pigmentation in the skin. Following extraction of tyrosinase inhibitors from edible vegetables or fruits, we found that the Prunus persica flesh extract (PPFE) exhibited potential inhibitory activity for melanogenesis. PPFE showed tyrosinase inhibitory activity in an enzymatic assay and PPFE also significantly inhibited the melanin formation in cultured mouse melan-a cells. Moreover, real-time RT-PCR analysis revealed that the inhibition of melanin production by PPFE was closely related to marked suppression of mRNA expression of tyrosinase and tyrosinase-related protein-1 and -2 (TRP-1 and TRP-2) in melan-a cells. Further investigation found that the modulation of tyrosinase expression by PPFE was associated with the transcriptional regulation of the microphthalmia-associated transcription factor (MITF). PPFE inhibited the promoter activity of MITF and suppressed MITF mRNA expression in melan-a cells. These results indicate that PPFE down-regulates melanogenesis-associated gene expression through MITF-mediated transcriptional regulation and these events might be related to the hypopigmentary effects of PPFE.

• Journal of Nutrition and Health
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• v.29 no.1
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• pp.97-103
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• 1996

This study was performed to investigate the effects of lipid on cellular senescence, lipid peroxide production, and morphological changes. For this study we used primary skin fibroblasts from neonate rats grown in media various lipid contents. Fibroblasts were cultured until they lost their proliferation potential either in control medium (Dulbecco's modified Eagle's medium supplement with 10% fetal bovine serum) or in media supplemented with various concentrations of lipid-cholesterol rice component from bovine serum. Cumulative population doublings(CPD, as an index of cellular life span), and cellular thiobarbituric acid reactive substances (TBARS, as an index of lipid peroxide) concentrations were measured and morphological changes were observed. CPD were shortened with increasing lipid concentration in media ; 28.12 for cells grown in control medium and 13.42, 11.42, and 6.19 for those grown in 0.1%, 1% and 5% lipid rich components containing media, respectively. Cellular proliferation ratios were those grown in 5% lipid rich components containing media were delayed and they were degenerated soon. TBARS concentrations were increased with increasing concentration of lipid in media. Morphological changes were observed in cells grown in control medium by cellular senescence. Especially lipid droplets were observed in cells grown in 5% lipid rich components containing media. Therefore it seems that lipid contents in media had an effect on cellular proliferation and cellular life span, possibly via lipid peroxide production.

• Toxicological Research
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• v.13 no.4
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• pp.393-400
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• 1997

The effects of superoxide dismutase(SOD) and catalase on the toxicity of paraquat(PQ) were studied using diethyldithiocarbamate(DDC), 3-amino-1,2,4-triazole(AT) which are inhibitors of Cu, Zn-SOD and catalase in rats. Sprague Dawley rats were divide into 6 groups: control, DDC, PQ, AT, DDC+PQ, and AT+PQ group. The PQ (50 mg/kg body weight(BW); about half dose of $LD_{50}$) was administered with orally, otherwise AT(1.0g/kg BW) and DDC(1.0g/kg BW) were administered by intrperitoneal(iP) injection. The survival rate of rats in PQ+AT group was significantly decreased compared with PQ group while the difference of survival rate between DDC group and DDC+PQ group was not significant. The SOD activity after administration of DDC was decreased in liver, lung and kidney, but catalase activity was not changed. The catalase activity in liver, lung and kidney of AT treated rats was decreased, while SOD activity was not changed in this group. The effects of DDC and AT to the PQ toxicity was also observed in primary cultured rat Skin fibroblasts. The viable cells that was measured with MTT method, was decreased in AT+PQ treated group compared to PQ treated group, but the difference of cell viability between DDC treat group and DDC+PQ treated group was not observed. This result, AT potentlate PQ toxicity while DDC were not affect, suggested that the decreased catalase activity lead to elevation of hydrogen peroxide levels and PQ toxicity may be correlate with the hydrogen peroxide rather than the superoxides.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.4
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• pp.430-436
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• 2010

The objective of the present studies was to develop and validate a system for isolation, purification and extended culture of pigment-producing cells in alpaca skin (melanocytes) responsible for coat color and to determine the effect of alpha melanocyte stimulating hormone treatment on mRNA expression for the melanocortin 1 receptor, a key gene involved in coat color regulation in other species. Skin punch biopsies were harvested from the dorsal region of 1-3 yr old alpacas and three different enzyme digestion methods were evaluated for effects on yield of viable cells and attachment in vitro. Greatest cell yields and attachment were obtained following dispersion with dispase II relative to trypsin and trypsin-EDTA treatment. Culture of cells in medium supplemented with basic fibroblast growth factor, bovine pituitary extract, hydrocortisone, insulin, 12-O-tetradecanolphorbol-13-acetate and cholera toxin yielded highly pure populations of melanocytes by passage 3 as confirmed by detection of tyrosinase activity and immunocytochemical localization of melanocyte markers including tyrosinase, S-100 and micropthalmia-associated transcription factor. Abundance of mRNA for tyrosinase, a key enzyme in melanocyte pigment production, was maintained through 10 passages showing preservation of melanocyte phenotypic characteristics with extended culture. To determine hormonal responsiveness of cultured melanocytes and investigate regulation of melanocortin 1 receptor expression, cultured melanocytes were treated with increasing concentrations of ${\alpha}$-melanocyte stimulating hormone. Treatment with ${\alpha}$-melanocyte stimulating hormone increased melanocortin receptor 1 mRNA in a dose dependent fashion. The results demonstrated culture of pure populations of alpaca melanocytes to 10 passages and illustrate the potential utility of such cells for studies of intrinsic and extrinsic regulation of genes controlling pigmentation and coat color in fiber-producing species.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.1
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• pp.79-83
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• 2004

In order to investigate the beneficial effects of parsely (Petroselinurn sativum) extract on skin, we measured the synthesis of total collagen and type I procollagen in cultured normal human fibroblast (NHF), the synthesis of prostaglandin E$_2$(PCE$_2$), interleukin 1 ${\alpha}$(IL -1 ${\alpha}$) and tumor necrosis factor ${\alpha}$ (TNF ${\alpha}$) in HaCaT cell and we also measured dermal thickness and density in hairless mouse (Female albino hairless mice, Skh:hr-1). As the results, the synthesis of total collagen and type I procollagen were increased 23％ and 18％ respectively, after 1 $\mu\textrm{g}$/mL parsley extract treatment. The producions of PGE$_2$ induced by UVB irradiation were decreased 60％ after 1 $\mu\textrm{g}$/mL parsley extract treatment. The treatment with 1 $\mu\textrm{g}$/mL parsley extract also decreased the synthesis of IL -1 ${\alpha}$ and TNF ${\alpha}$ induced by 10 uM RA, 100 $\mu\textrm{g}$/mL SLS and 30 mJ/$\textrm{cm}^2$ UVB irradiation, After 4 days treatment with 1％ parsley extract, the dermal thickness of hairless mouse was increased 1.5 times and the density of dermis was tighter than control. These results indicate that parsley extract have anti-aging and anti-irritation effects on skin.