• Journal of Industrial and Engineering Chemistry
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• 제68권
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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.

• 대한화장품학회지
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• 제49권1호
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• pp.47-58
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• 2023

마이크로바이옴의 화장품 응용에 대한 관심이 높아지고 마이크로바이옴 추출물의 복합적인 효능에 의한 피부 개선 연구가 중요해지고 있다. 본 연구에서는 세포 및 3차원 배양 인공피부를 이용한 in vitro 시험법을 통하여 마이크로바이옴 중 피부 주름 개선 효능이 뛰어난 Bifidobacterium bifidum (B. bifidum)을 선정하였고, 이의 추출물을 함유한 화장품의 피부주름개선 효능평가를 진행하였다. 이후, 비피다발효여과물 원료의 세포독성 및 항산화, 항염 및 주름개선 유효성 시험을 실시하였다. 세포독성이 없고 효능이 우수한 5% (v/v) 농도의 비피다발효여과물이 함유된 에멀젼을 제조하였고 placebo 에멀젼와 비교하여 총 21 명 여성을 대상으로 2 주간 임상시험을 수행하였다. Antera 3D를 사용하여 눈가 부위의 주름을 기기 평가하였다. 주름 개선도는 placebo 에멀젼 사용군 대비하여 주름 감소율(%)에서 5.6% 차이를 보였다. 이러한 3차원 배양 인공피부를 활용한 마이크로바이옴 추출물 선정 및 이를 함유한 화장품의 효능 연구는 다양한 효능을 가지는 마이크로바이옴 화장품의 개발 및 메커니즘 연구에 응용될 수 있을 것이라 기대된다.

• Archives of Pharmacal Research
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• 제28권7호
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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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• 제28권12호
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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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• 제18권4호
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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.

• 한국식품저장유통학회지
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• 제24권2호
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• pp.320-324
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• 2017

본 연구에서 루꼴라(Eruca sativa)에 대한 다양한 생리활성을 조사하여 기능성소재 응용가능성을 검토하였다. 루꼴라 추출물은 B. subtilis, E. coli, C. albicans에 대한 항균활성이 매우 우수하게 나타났으며 특히 그람양성세균 B. subtilis에 대한 항균활성이 높았다. 또한, 인볼루크린 발현에 대한 면역조직화학 분석을 인공피부를 이용하여 분석한 결과 루꼴라 추출물을 처리(50 mg/L)한 경우에 0.1% DMSO로 처리한 대조군에 비하여 인볼루크린 발현이 뚜렷하게 증가하였으며 동일농도로 처리된 합성물질 WY14643과 비교하였을 때에도 비슷한 발현양상을 보여 주었다. 이와 같은 결과로 미루어 볼 때 루꼴라 추출물은 피부염 개선 및 피부장벽 기능을 향상할 수 있는 천연물 기반의 소재로 활용하기 위한 매우 효과적인 재료가 될 수 있다고 사료된다.

• Biomolecules & Therapeutics
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• 제32권2호
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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.

• 대한화장품학회지
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• 제39권1호
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• pp.31-38
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• 2013

피부 자극과 부식 검사에 사용되는 동물실험을 대체하기 위한 방법으로 인공피부가 개발되어 왔다. 최근에 본 연구진은 녹각교를 함유하는 새로운 인공피부를 구축하였다. 현재 연구에서는 녹각교를 포함하는 인공피부를 사용하여 물질의 독성도 검사를 수행하였다. 그리하여 sodium dodecylsulfate (SDS) 또는 sodium carbonate를 인공피부에 도포하였고 표피의 손상 정도를 H&E와 면역조직화학 염색을 통하여 평가하였다. 인공피부의 표피는 SDS와 sodium carbonate에 의해 농도 의존적으로 영향을 받았다. 더 나아가 이들 물질에 의하여 p63의 발현이 감소하였다. 그러므로 녹각교를 함유하는 인공피부는 동물실험을 대체할 수 있는 모델로써 사용될 수 있고 in vitro에서 자극과 부식 검사 시험법의 발전에 도움을 줄 것으로 예상된다.

• 대한화장품학회지
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• 제29권2호
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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).

• 대한화장품학회지
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• 제38권3호
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• pp.219-224
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• 2012

최근 유럽연합에서는 동물실험을 통하여 검증된 성분을 함유하고 있는 화장품에 대하여 판매를 금지할 것을 선언하였다. 그리하여 동물실험을 대체할 동물실험 대체모델의 개발이 필요해졌다. 인공피부는 화장품, 의약품 및 의료기기의 안전 테스트에 있어서 아주 중요한 시스템이다. 본 연구에서는 기저막과 표피를 가지고 있는 최적의 인공피부를 만들기 위한 시도를 하였다. 이러한 목적으로 중간엽줄기세표(MSCs, mesenchymal stem cells)와 지방전구세포를 진피세포인 섬유모세포와 혼합하여 진피대체물을 만들었다. 기저막과 표피의 형성은 면역조직화학 염색(immunohistochemical stains)을 통하여 확인하였다. 여러 가지 모델 중에서, 중간엽줄기세포를 혼합한 모델에서 표피의 두께가 제일 두꺼웠으며 또한 PCNA와 involucrin의 분포가 실제 사람피부와 비슷하였다. 결론적으로, 본 연구의 결과는 진피대체물에 중간엽줄기세포를 혼합한 인공피부가 동물실험 대체모델로 개발될 수 있다는 점을 제시한다.