• Journal of Industrial and Engineering Chemistry
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• v.68
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• pp.238-245
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• 2018

The skin experiences constant physical stimuli, such as stretching. Exposure to excessive physical stimuli stresses the skin and can accelerate aging. In this study, we applied a method that allowed human fibroblasts and keratinocytes to be perfused with media to form 3D skin equivalents that were then uniaxially 10%-stretched for 12 h per day (at either 0.01 or 0.05 Hz) for up to 7 days to form wrinkled skin-on-a-chip (WSOC). There was more wrinkling seen in skin equivalents under 0.01 Hz uniaxial stretching than there was for non-stretched skin equivalents. At 0.05 Hz, the stratum corneum almost disappeared from the skin equivalents, indicating that stretching was harmful for the epidermis. At both frequencies, the production of collagen and related proteins in the skin equivalents, such as fibronectin 10 and keratin, decreased more than those in the non-stretched equivalents, indicating that the dermis also suffered from the repeated tensile stress. These results suggest that WSOCs can be used to examine skin aging and as an in vitro tool to evaluate the efficacy of anti-wrinkle cosmetics and medicines.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.49 no.1
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• pp.47-58
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• 2023

Recently, along with the remarkable increase in interest in microbiome cosmetics, the application of microbiome extracts in the complex efficacy as anti-aging, brightening etc. has become very important. In this study, Bifidobacterium bifidum (B. bifidum), which has excellent anti-wrinkle efficacy among the microbiome, was selected through an in vitro test using cells and 3D human skin equivalents. And the anti-wrinkle efficacy of cosmetics containing B. bifidum was evaluated by clinical test. Thereafter, the cytotoxicity, anti-oxidation, anti-inflammatory and anti-wrinkle efficacy of the of the bifida fermented filtrate were tested. An emulsion containing bifida fermented filtrate at a concentration of 5% (v/v) with no cytotoxicity and excellent efficacy was prepared with the placebo emulsion. The clinical test was conducted on a total of 21 women at 2 weeks comparing the bifida emulsion and placebo emulsion. Wrinkles around the eyes were instrumentally evaluated using Antera 3D. The wrinkle reduction rate of the Bifida emulsion group compared with the placebo emulsion group differed by 5.6%. Overall, the selection of microbiome using 3D human skin equivalents and the efficacy study of cosmetics with the microbiome extracts would be actively studied to the development of microbiome cosmetics and skin microbiome mechanisms.

• Archives of Pharmacal Research
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• v.28 no.7
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• pp.784-790
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• 2005

In this study, we evaluate the effects of (-)-epigallocatechin-3-gallate (EGCG) on ultraviolet B(UVB)-irradiated living skin equivalents (LSEs). Histologically, UVB irradiation induced thinning of the LSE epidermis, whereas EGCG treatment led to thickening of the epidermis. Moreover, EGCG treatment protected LSEs against damage and breakdown caused by UVB exposure. Immunohistochemically, UVB-exposed LSEs expressed p53, Fas, and 8-hydroxy-deoxyguanosine (8-OHdG), all of which are associated with apoptosis. However, EGCG treatment reduced the levels of UVB-induced apoptotic markers in the LSEs. In order to determine the signaling pathways induced by UVB, Western blot analysis was performed for both c-Jun $NH_2$-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK), which are associated with UVB-induced oxidative stress. UVB activated JNK in the epidermis and dermis of the LSEs, and EGCG treatment reduced the UVB-induced phosphorylation of JNK. In addition, p38 MAPK was also found to have increased in the UVB-exposed LSEs. Also, EGCG reduced levels of the phosphorylation of UVB-induced p38 MAPK. In conclusion, pretreatment with EGCG protects against UVB irradiation via the suppression of JNK and p38 MAPK activation. Our results suggest that EGCG may be useful in the prevention of UVB-induced human skin damage, and LSEs may constitute a potential substitute for animal and human studies.

• Journal of Microbiology and Biotechnology
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• v.28 no.12
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• pp.2121-2132
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• 2018

Abnormal melanin synthesis results in several hyperpigmentary disorders such as freckles, melanoderma, age spots, and other related conditions. In this study, we investigated the anti-melanogenic effects of an extract from the microalgae Chlamydomonas reinhardtii (CE) and potential mechanisms responsible for its inhibitory effect in B16F10, normal human epidermal melanocyte cells, and human skin-equivalent models. The CE extract showed significant dose-dependent inhibitory effects on ${\alpha}$-melanocyte-stimulating, hormone-induced melanin synthesis in cells. Additionally, the CE extract exhibited suppressive effects on the mRNA and protein expression of microphthalmia-associated transcription factor, tyrosinase, tyrosinase-related protein-1, and tyrosinase-related protein-2. The CE extract also inhibited the phosphorylation of protein kinase A and extracellular signal-related kinase, which function as upstream regulators of melanogenesis. Using a three-dimensional, reconstructed pigmented epidermis model, the CE-mediated, anti-pigmentation effects were confirmed by Fontana-Masson staining and melanin content assays. Taken together, CE extract can be used as an anti-pigmentation agent.

• The Korean Journal of Physiology and Pharmacology
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• v.18 no.4
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• pp.327-331
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• 2014

In this study we investigated the effects of fucoidan on the proliferation of fibroblasts and the reconstruction of a skin equivalent (SE). Fucoidan significantly stimulated the proliferation of CCD-25Sk human fibroblasts and Western blot analysis demonstrated that fucoidan markedly increased the expression of cyclin D1 and decreased the expression of p27. Fucoidan was used to reconstruct SE. Immunohistochemical staining showed that the addition of fucoidan to dermal equivalents increased expression of proliferating cell nuclear antigen (PCNA) and p63. In addition, expression of ${\alpha}6$-integrin was significantly increased by fucoidan, whereas expression of ${\beta}1$-integrin, type 1 collagen, elastin, fibronectin did not markedly change. These results suggest that fucoidan has positive effects on epidermal reconstruction and will therefore be beneficial in the reconstruction of SE.

• Food Science and Preservation
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• v.24 no.2
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• pp.320-324
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• 2017

Eruca sativa is a rocket plant and a member of the Brassicaceae, which is considered to be an important chemo-preventive plant family. Although Eruca sativa has positive biological effects, the effect of Eruca sativa extract (ES) on improvement of skin barrier function has not been reported. In this study, we investigated the applicability of functional materials by examining a variety of physiological activities of Eruca sativa extract. ES showed anti-microbial activities against Bacillus subtilis, Escherichia coli, and Candida albicans. In particular, antimicrobial activities of ES against B. subtilis was the highest. Additionally, immunohistochemical analysis of protein marker related to keratinocyte differentiation was determined. The treatment by ES (50 mg/L) showed a significant increase of involucrin expression compared with treatment by 0.1% DMSO as a control in skin equivalents, the ES-treated group showed similar level in the expression of involucrin compared to the group treated with the same concentration of WY14643 in $EpiDerm^{TM}$, a three-dimensional model of skin equivalents. These results indicate that ES promotes the expression of protein related to barrier properties of the skin. Therefore, ES may be an effective ingredient for skin barrier improvement.

• Biomolecules & Therapeutics
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• v.32 no.2
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• pp.224-230
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• 2024

Pinitol (3-O-Methyl-D-chiro-inositol) has been reported to possess insulin-like effects and is known as one of the anti-diabetic agents to improve muscle, liver, and endothelial cells. However, the beneficial effects of pinitol on the skin are not well known. Here, we investigated whether pinitol had effects on human dermal fibroblasts (HDFs), and human dermal equivalents (HDEs) irradiated with ultraviolet A (UVA), which causes various damages including photodamage in the skin. We observed that pinitol enhanced wound healing in UVA-damaged HDFs. We also found that pinitol significantly antagonized the UVA-induced up-regulation of matrix metalloproteinase 1 (MMP1), and the UVA-induced down-regulation of collagen type I and tissue inhibitor of metalloproteinases 1 (TIMP1) in HDEs. Electron microscopy analysis also revealed that pinitol remarkably increased the number of collagen fibrils with regular banding patterns in the dermis of UVA-irradiated human skin equivalents. Pinitol significantly reversed the UVA-induced phosphorylation levels of ERK and JNK but not p38, suggesting that this regulation may be the mechanism underlying the pinitol-mediated effects on UVA-irradiated HDEs. We also observed that pinitol specifically increased Smad3 phosphorylation, which is representative of the TGF-β signaling pathway for collagen synthesis. These data suggest that pinitol exerts several beneficial effects on UVA-induced damaged skin and can be used as a therapeutic agent to improve skin-related diseases.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.39 no.1
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• pp.31-38
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• 2013

To substitute animal test, skin equivalents (SEs) have been developed for skin irritation and corrosion test. Recently, we have developed new SEs containing Cervi cornus Colla (CCC). In the present study, we used the SEs for cutaneous cytotoxicity test. Sodium dodecylsulfate (SDS) or sodium carbonate was applied to the SEs-, and the epidermal damage by H&E and immunohistochemical stains was evaluated. Our results showed that SDS or sodium carbonate affected the epidermal part of SEs containing CCC in a dose-dependent manner and decreased the expression of p63. It is concluded that SEs containing CCC could be used for an alternative model of animal test and would be greatly helpful in the development of in vitro irritation and corrosion test.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.29 no.2 s.43
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• pp.105-130
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• 2003

Human skin equivalents, also designated as cultured skin substitute (Boyce and Warden, 2002) or organotypic co-cultures (Maas-Szabowski et al., 1999, 2000, 2003), are three-dimensional systems that are engineered by seeding fibroblasts into a three-dimensional dermal matrix. Such a dermal equivalent is then subsequently seeded with human keratinocytes. After cell attachment, the culture is kept first under submerged condition to allow keratinocyte proliferation. Thereafter, the culture is lifted the air-liquid interface (A/L) to expose the epidermal compartment to the air, and to further induce keratinocyte differentiation. During the air-exposure, nutrients from the medium will diffuse through the underlying dermal substrate towards the epidermal compartment and support keratinocyte proliferation and differentiation. Under these conditions, a HSE is formed that shows high similarity with the native tissue from which it was derived (Figure 1) (Bell et at., 1981; Boyce et al., 1988; Ponec et al., 1997;El Ghalbzouri et al.., 2002).

• 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.