• Microbiology and Biotechnology Letters
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• v.49 no.4
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• pp.485-492
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• 2021

In this study, the whitening effect of Abelmoschus esculentus extract was investigated to confirm its applicability in cosmetics. To determine the whitening effect, the tyrosinase-inhibitory activity of Abelmoschus esculentus hot water extract (AEWE) and 70% ethanol extract (AEEE) was measured. At the final concentration of 1000 ㎍/ml, AEWE showed an inhibitory activity of 22.2% and AEEE of 32.8%. To determine the whitening effect at the cellular level, the viability of melanoma cells treated with AEWE and AEEE was evaluated using the MTT assay. At concentrations of 100 ㎍/ml or less, both AEWE- and AEEE-treated groups showed cell survival rates of >95%. Furthermore, in both AEWE- and AEEE-treated melanoma cells, the melanin content decreased in a concentration-dependent manner. The inhibitory effects of AEWE and AEEE used at 5, 10, 50, and 100 ㎍/ml on protein expression were measured by western blot, with β-actin as the positive control. At a concentration of 100 ㎍/ml, AEWE showed an inhibitory effect of 88.1%, 24.8%, 62.2%, and 42.9% on microphthalmia-associated transcription factors (MITF), tyrosinase, tyrosinase-related proteins (TRP)-1, and TRP-2 factors, respectively. At the same concentration, AEEE showed inhibitory effect of 65.3%, 58.3%, 66.2%, and 65.3% against MITF, tyrosinase, TRP-1, and TRP-2 factors, respectively. In conclusion, the whitening effects of AEWE and AEEE were verified, and their applicability as a natural ingredient in cosmetics was confirmed.

• Microbiology and Biotechnology Letters
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• v.48 no.4
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• pp.556-563
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• 2020

The purpose of this study was to verify the antioxidant and whitening effects of fermented Ambrosia trifida L. extract (ATFE) and to verify its usefulness as a cosmetic material. The antioxidant effects were measured by assessing the electron-donating capacity and 2,2-azino-bis(3-ethyl-benthiazoline-6-sulfonic acid) (ABTS) radical scavenging ability of these extracts. ATFE was shown to have an electron-donation capacity of 68.4% at a concentration of 1000 ㎍/ml. While its ABTS+ radical scavenging ability was shown to be 58.7% at the same concentration. The ATFE tyrosinase inhibitory effect, which is related to skin-whitening, was shown to be 32.35% at a concentration of 1000 ㎍/ml and a cell viability assay using melanoma cells showed a 14.8% reduction in cell viability at a concentration of 100 ㎍/ml. Surviving cells were then used in western blot analyses to evaluate the protein inhibitory effects of ATFE at 25, 50, 100 ㎍/ml where β-actin was used as a positive control. The whitening effects of these extracts were also evaluated by western blot and show that the expression of microphthalmia-associated transcription factors, Tyrosinase-related proteins (TRP)-1, TRP-2 and Tyrosinase were all inhibited, 51.14%, 55.4%, 38.6%, 83.77% respectively, at 100 ㎍/ml ATFE. The efficacy of the whitening effects was verified and the suitability of ATFE as a cosmetic material was assured.

• Journal of Microbiology and Biotechnology
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• v.28 no.3
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• pp.375-380
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• 2018

We have previously found that mycelia culture broth of eight kinds of traditional herbal extracts fermented with Phellinus linteus (previously named as 8-HsPLCB) not only inhibited melanin and tyrosinase activity, but also reduced the contents of melanogenesis-related proteins, including tyrosinase and microphthalmia-associated transcription factor, in 3-isobutyl-1-methylxanthine-stimulated B16F0 melanoma cells. For a further study, the effect of 8-HsPLCB against skin pigmentation in brown guinea pigs with ultraviolet B (UVB)-induced hyperpigmentation was investigated. 8-HsPLCB (3%) and arbutin (2%) as positive controls were applied topically twice daily for 4 weeks to the hyperpigmented areas. 8-HsPLCB showed skin-lightening effect as effective as arbutin, one of the most widely used in whitening cosmetics. Melanin index values as the degree of pigmentation showed a significant reduction week by week post 8-HsPLCB treatment and then substantially reduced by 4 weeks. The degree of depigmentation after 4 weeks of topical application with 8-HsPLCB was 32.2% as compared with before treatment (0 week). Moreover, using Fontana-Masson staining and hematoxylin-eosin staining, 8-HsPLCB reduced melanin pigmentation in the basal layer of the epidermis and epidermal thickness changes exposed to the UV-B irradiation as compared with non-treatment and vehicle treatment. The intensity of the skin-lightening effect of 8-HsPLCB was similar to arbutin. These results suggest that the skin-lightening effect of 8-HsPLCB might be resulted from inhibition of melanin synthesis by tyrosinase in melanocytes. To conclude, 8-HsPLCB treatment showed reduction of the melanin pigment and histological changes induced by UV irradiation in brown guinea pigs.

• Journal of Life Science
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• v.28 no.7
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• pp.802-810
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• 2018

Aging is accompanied by changes in the body, such as graying hair, wrinkles, and black spots composed of lipid peroxides and proteins. Melanin is a polymer substance produced by an oxidation polymerization reaction from tyrosine, and it determines the color of hair and skin. It has been reported that melanin is synthesized by melanocyte, and its excessive production by reactive oxygen species is associated with aging. The purpose of this study was to determine the direct effects of Musa paradisiaca peel ethanolic extract (MPEE) on antioxidative activity and melanin synthesis. It was observed that the antioxidant activity of MPEE was similar to that of vitamin C, a positive control, in both DPPH radical scavenging assay and reducing power assay. In order to examine cytotoxicity prior to cell experimentation, 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed for B16F1 cells. MPEE was not cytotoxic at $32{\mu}g/ml$ or less. In addition, MPEE increased melanin synthesis in live cells in addition to tyrosinase activity and melanin synthesis in dihydroxyphenylalanine (DOPA)-oxidation assay in vitro. Moreover, MPEE increased melanin synthesis in cells aged by pretreatment with $H_2O_2$. The expression levels of tyrosinase-related protein (TRP)-1, TRP-2, and superoxide dismutase (SOD)-2 by western blot analysis were increased in the presence of MPEE. These results suggest that MPEE could promote the melanin synthesis as an antioxidative substance.

• Journal of the Korean Applied Science and Technology
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• v.35 no.2
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• pp.325-335
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• 2018

This study is for checking the possibility of ginseng complex as cosmetic materials. For this we carried out biological active evaluation about anti-inflammatory and whitening effects by using ethanol extract of ginseng complex. Samples were prepared by extracting 70% ethanol from each of Panax ginseng C. A. Meyer (A), Phellinus linteus (B) and Pinus rigida Mill. (C), and mixing them at a ratio of (A) 1 : (B) 1 : (C) 0.5. In order to evaluate the anti-inflammatory effects of the samples in macrophages (RAW 264.7 cells), MTT assay was used to evaluate the toxicity of the samples and the inhibitory activity of nitric oxide production and the expression levels of inflammation-related proteins and genes. To evaluate the whitening effect of the samples in melanoma (B16F10 cell), MTT assay was used to evaluate the toxicity of the sample, cellular tyrosinase inhibition, and melanin contents. The inhibitory activity of nitric oxide in the LPS-induced RAW 264.7 cells was 71.2% at $25{\mu}g/mL$ concentration and western blot analysis showed that the expression of iNOS and COX-2 protein decreased in a concentration-dependent manner. Inhibition of tyrosinase activity showed 36.8% inhibition at $50{\mu}g/mL$ concentration of ginseng complex and inhibition of melanin contents showed 47.8% inhibition at $50{\mu}g/mL$ concentration. From the results of the experiment, it was confirmed that the ginseng complex had excellent anti-inflammatory and whitening effect and could be used as a safe natural cosmetic material in the future.

• Molecules and Cells
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• v.44 no.10
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• pp.736-745
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• 2021

Although various marine ingredients have been exploited for the development of cosmetic products, no previous study has examined the potential of seaweed extracellular vesicles (EV) in such applications. Our results revealed that EV from Codium fragile and Sargassum fusiforme effectively decreased α-MSH-mediated melanin synthesis in MNT-1 human melanoma cells, associated with downregulation of MITF (microphthalmia-associated transcription factor), tyrosinase and TRP1 (tyrosinase-related proteins 1). The most effective inhibitory concentrations of EV were 250 ㎍/ml for S. fusiforme and 25 ㎍/ml for C. fragile, without affecting the viability of MNT-1 cells. Both EV reduced melanin synthesis in the epidermal basal layer of a three-dimensional model of human epidermis. Moreover, the application of the prototype cream containing C. fragile EV (final 5 ㎍/ml) yielded 1.31% improvement in skin brightness in a clinical trial. Together, these results suggest that EV from C. fragile and S. fusiforme reduce melanin synthesis and may be potential therapeutic and/or supplementary whitening agents.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.11
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• pp.1695-1700
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• 2014

Oriental herbal liquor (Yakju) is a type of Korean traditional alcoholic beverage that uses Nuruk and oriental herbs for fermentation. The purpose of this study was to develop cosmetic ingredients using Jubak, which is a by-product of alcoholic fermentation of oriental herbal liquor. To investigate antioxidant, whitening, and anti-aging effects of Jubak, we prepared extract of Jubak and its solvent fractions. Ethyl acetate fraction (KSD E4-3) showed the most prominent free radical [1,1-diphenyl-2-picrylhydrazyl (DPPH)] scavenging activity ($SC_{50}$: 0.75 mg/mL). KSD E4-3 significantly inhibited in vitro mushroom tyrosinase activity ($IC_{50}$: 0.82 mg/mL) and reduced the melanin contents in mouse melanoma melanocyte, B16F10 cells. KSD E4-3 down-regulated protein expression of tyrosinase related proteins (TRP)-1, -2, which play key roles in melanogenesis. For anti-aging effects, inhibition of matrix metalloproteinase (MMPs) expression was evaluated using human keratinocyte, HaCaT cells. Treatment of HaCaT cells with KSD E4-3 reduced expression of MMP-1, -2, -9 and inhibited proteolytic activities of MMP-2, -9. These results suggest that KSD E4-3 induces down-regulation of cellular melanogenesis and protects against photoaging induced by UVB-induced damage. Thus KSD E4-3 could potentially be a valuable cosmetic ingredient.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2007.11a
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• pp.79-92
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• 2007

Oxidative stress have known to be a risk factor for the degenerative processes and closely related to a lot of diseases. It is well established that antioxidants are good in protection and therapeutic means against oxidative damage. There is increasing interest in natural antioxidants and many natural antioxidants have been found and utilized as the possible protection for various diseases and skin aging. We have screened natural antioxidant agents for cosmeceuticals, nutraceuticals, and drugs as therapeutic and preventive means against oxidative stress, and have developed a number of novel antioxidants from various natural sources. A novel melanin synthesis inhibitor, Melanocin A, isolated from the metabolite of a fungal strain Eupenicillium shearii F80695 inhibited mushroom tyrosinase and melanin biosynthesis of B16 melanoma cells with $IC_{50}$ value of 9.0 nM and MIC value of $0.9\;{\mu}M$, respectively. Melanocin A also exhibited potent antioxidant activity by scavenging of DPPH and superoxide anion radicals. UV was found to increase the level of hydrogen peroxides and other reactive oxygen species (ROS) in skin tissues. This increase in ROS may not only alter the structure and function of many genes and proteins directly but may also modulate their expressions through signal transduction pathways and, ultimately, lead to skin damage. We investigated the effect of Melanocin A on UV-induced premature skin aging. Firstly, the effect of Melanocin A on UV-induced matrix metalloproteinase (MMP)-9 expression in an immortalized human keratinocyte cell line, HaCaT in vitro was investigated. Acute UV irradiation induced MMP-9 expression at both the mRNA and protein levels and Melanocin A suppressed this expression in a dose-dependent manner. We then investigated UV-induced skin changes in hairless mice in vivo by Melanocin A. Chronic exposure of hairless mouse dorsal skin to UV increased skin thickness and induced wrinkle formation and the gelatinase activities of MMP-2 and MMP-9. Moreover, Melanocin A significantly suppressed UV-induced morphologic skin changes and MMP-2 and MMP-9 expression. These results show that Melanocin A can prevent the harmful effects of UV that lead to skin aging. Therefore, we suggest that Melanocin A should be viewed as a potential therapeutic agent for preventing and/or treating premature skin aging. Terrein is a bioactive fungal metabolite isolated from Penicillium species. Terrein has a relatively simple structure and can be easily synthesized. However, the biologic effects of terrein are comparatively unknown. We found for the first time that terrein potently inhibit melanin production in melanocytes and has a strong hypopigmentary effect in a spontaneously immortalized mouse melanocyte cell line, Mel-Ab. Treatment of Mel-Ab cells with terrein (10-100 mM) for 4 days significantly reduced melanin levels in a dose-dependent manner. In addition, terrein at the same concentration also reduced tyrosinase activity. We then investigated whether terrein influences the extracellular signal-regulated protein kinase (ERK) pathway and the expression of microphthalmia-associated transcription factor (MITF), which is required for tyrosinase expression. Terrein was found to induce sustained ERK activation and MITF down-regulation, and luciferase assays showed that terrein inhibits MITF promoter activity in a dose-dependent manner. To elucidate the correlation between ERK pathway activation and a decreased MITF transcriptional level, PD98059, a specific inhibitor of the ERK pathway, was applied before terrain treatment and found to abrogate the terrein-induced MITF attenuation. Terrein also reduced the tyrosinase protein level for at least 72 h. These results suggest that terrain reduces melanin synthesis by reducing tyrosinase production via ERK activation, and that this is followed by MITF down-regulation.

• Microbiology and Biotechnology Letters
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• v.47 no.2
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• pp.195-200
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• 2019

The purpose of this study was to investigate the melanogenesis inhibiting activity of the ethanol extract from Polygonum amphibium L. Firstly, the n-hexane (Hx), chloroform ($CHCl_3$), ethyl acetate (EA), n-butanol (BuOH), and water (Water) fractions were isolated from the P. amphibium L. ethanol extract. The efficacy of melanogenesis was found to significantly decrease via the EA and BuOH fractions when compared to the control in B16F10 cells. EA particularly showed the lowest melanin content in B16F10 cells when compared to all the other extracts. Concentration-dependent inhibition of melanin synthesis was also observed in the EA fraction at concentrations below $50{\mu}g/ml$, which did not exhibit cytotoxicity in B16F10 cells. Notably, the expression of three key proteins (tyrosinase, tyrosinase-related protein-1 (TRP-1), and TRP-2), which are involved in melanogenesis, were significantly decreased via the EA fraction. EA also inhibited body pigmentation in vivo in a zebrafish model. Overall, we demonstrated melanogenesis suppression using the EA fraction from P. amphibium L., which could be a potential candidate for an antimelanogenesis agent.

• Microbiology and Biotechnology Letters
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• v.42 no.2
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• pp.106-113
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• 2014

For the development of high value consumables utilizing jujube and grape, we investigated the biological activities of a variety of existing fermentation products of jujube and grape. The results revealed that ethanol fermentation products of jujube and grape had a higher antioxidative activity than acetic acid fermentation products. In addition, the ethanol fermentation products of jujube (JEF) had the highest antioxidative activity, with it being greater than that of the ethanol fermentation products of grape (GEF), the acetic acid fermentation products of jujube (JAF) and the acetic acid fermentation products of grape (GAF). As regards tests on whitening effects, JEF exhibited the highest tyrosinase inhibition effects amongst the test groups. However, when immunofluorecence was employed, JAF was seen to inhibit the expression of proteins related to the whitening effect. In the lipopolysaccharide-stimulated mode peritoneal macrophage model, all tested groups of fermentation products (JEF, GEF, JAF and GAF) suppressed nitric oxide production dose-dependently, with ethanol fermentation products demonstrating a higher nitric oxide expression inhibition effect than acetic acid fermentation products. When subjected to antibacterial activity tests, GAF exhibited antibacterial activity against all tested strains except Propionibacterium acnes. Both GAF and JEF revealed high antibacterial activity against Escherichia coli.