• Biomolecules & Therapeutics
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• v.32 no.2
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• pp.231-239
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• 2024

Methyl anthranilate (MA) is a botanical fragrance used in food flavoring with unexplored potential in anti-pigment cosmetics. MA dose-dependently reduced melanin content without affecting cell viability, inhibited dendrite elongation and melanosome transfer in the co-culture system of human melanoma cells (MNT-1) and human keratinocyte cell line (HaCaT), and downregulated melanogenic genes, including tyrosinase, tyrosinase-related protein 1 and 2 (TRP-1, TRP-2). Additionally, MA decreased cyclic adenosine monophosphate (cAMP) production and exhibited a significant anti-pigmentary effect in Melanoderm^TM. These results suggest that MA is a promising anti-pigmentary agent for replacing or complementing existing anti-pigmentary cosmetics.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.31 no.2
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• pp.143-149
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• 2005

Objects : With the advancement of tissue engineering techniques, the effort to develop bioartificial mucosa have been actively delivered. The problem we met with this technique is the lack of mechanical strength between kerationocyte layer and dermal layer, where in the normal skin and mucosa, they are tightly bound with rete ridge structure. The purpose of this study is to understand the 2D and 3D structure of rete ridge of mucosa and skin paddle for rendering more biomimetic structure to the artificial mucosa. Materials and Methods : Oral mucosa and skin from the patients who received the oral surgery and maxillofacial reconstruction were harvested. The epidermis was separated from the dermis after treating with dispase for 12-16 hours. H&E staining was performed for 2D(dimensional) structure study and confocal LASER and SEM study were performed for 3D structure. Mean height(Sc) and arithmetic mean deviation(Sa) of all surface height were calculated. Results : The average height of rete ridge of skin flap was between $67.14{\mu}m$ and $194.55{\mu}m$. That of oral mucosa was between $146.26{\mu}m$ and $167.51{\mu}m$. Pressure bearing area and attached gingiva of oral mucosa showed deeper rete ridges. Conclusion : To obtain the adequate strength of artificially cultured keratinocyte skin and mucosa flap, it is necessary to imitate the original skin and mucosa structure, especially rete ridge. Through this study, 2D and 3D rete ridge structure of normal mucosa and skin was obtained. These results can be used as basis for substrate morphology for keratinocytes culture.

• The Journal of the Convergence on Culture Technology
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• v.8 no.6
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• pp.849-854
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• 2022

In this study, the anti-inflammatory effect of supercritical carbon dioxide selective extracts, which extract decursin and decursinol angelate, the vital active ingredients of Angelica gigas Nakai, in high yield, was measured compared to that of ethanol extracts. To measure the anti-inflammatory effect, the production of nitric oxide(NO), an inflammatory mediator, and interleukin(IL)-6 and IL-8, inflammatory cytokines, was measured. NO production was measured by Griess assay on Raw 264.7 cells induced inflammatory response by lipopolysaccharide(LPS), and IL-6 and IL-8 production was measured by enzyme-linked immunoadsorbent assay(ELISA) on human keratinocyte cell line(HaCaT) cells induced inflammatory response by tumor necrosis factor(TNF)-α. The amount of NO production was suppressed outstandingly by the supercritical carbon dioxide extracts compared to the ethanol extracts. The amount of IL-6 and IL-8 production was increased by the ethanol extracts, whereas statistically significantly inhibited by supercritical carbon dioxide extracts at the concentration of 6.25 ㎍/mL(P<0.01). Through these results, we confirmed that the supercritical carbon dioxide selective extracts of Angelica gigas Nakai could be used as an anti-inflammatory cosmeceutical material to alleviate atopic dermatitis.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.1 s.49
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• pp.97-102
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• 2005

One of the key molecules involved in skin moisture is hyaluronan (hyaluronic acid, HA) with its associated water of hydration. The predominant component of the ECM (extracellular matrix) of skin is HA. It Is the primordial and the simplest of the GAGs (glycosaminoglycans), a water-sorbed macromolecule In extracellular matrix, Included between the vital cells of epidermis. In the skin, HA was previously thought to derive extlusively from dermis. But, recent studies revealed that HA could be synthesized in epidermis. Flavonoids are polyphenolic compounds that is found mainly in foods of plant origin. Kaempferol was known to increase glutathione synthesis in human keratinocyte. And quercetin blocked PPAR-meidated keratinocyte differentiation as lipoxygenase inhibitors. In this study, we sought to evaluate the effect of flavonid, kaempferol and quercetin on production HA in keratinocyte. We examined the changes of three human hyaluronan synthase genes (HASI, HAS2, HAS3) expression by semi-quantitative RT-PCR when kaempferol or quercetin was added to cultured human keratinocytes. We found that these flavonoids slightly upregulated HAS2, HAS3 mRNA after 24 h. And we investigated the effect on HA production by ELISA. When we evaluated the level of HA in culture medium after 24 h incubation. We found enhanced accumulation of HA in the culture medium. Although the effects of above flavonoids are less than retinoic acid, the data indicate that kaempferol, quercetin can dose-dependently increase the level of HA in epidermis cell line. It suggested that flavonoid, kaempferol, and quercetin increased production of HA in skin and it helped to elevate skin moisture and improve facial wrinkle.

• Environmental Mutagens and Carcinogens
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• v.24 no.2
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• pp.91-98
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• 2004

Although 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD) is a powerful carcinogen in several species, limited model system exist to study carcinogenicity of this compound at cellular level. To enhance our under-standing of carcinogenicity of TCDD at cellular level, we investigated micronucleus (MN) frequency as a index of genetic toxicity and whether TCDD can transform the human cells in culture. Normal human cell lines, skin keratinocyte HaCaT, Chang liver and breast MCF10A cells were used. TCDD did not affect the cell viability of the Chang liver, HaCaT and MCF10A cells. The frequency of micronucleus was increased after treatment of TCDD for 24hr in Chang liver and HaCaT cells, but not changed in MCF10A cells. And we observed putative transformed cells in Chang liver cells exposed to 1 $\mu$M TCDD for 2 weeks. The putative transformed cells were also observed in HaCaT cells with subsequent exposure to TCDD (0.1, 1, 10, 100 nM) for 2 weeks after initial exposure to MNNG, but not observed in MCF10A cells. Collectively, these results indicate that the ability of TCDD to induce micronuclei may be involved in cellular transformation of Chang liver and HaCaT cells. Our putative TCDD-transformed cells of Chang liver and HaCaT are expected to provide a clue to the elucidation of TCDD-induced transformation pathway.

• Genomics & Informatics
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• v.3 no.2
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• pp.66-72
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• 2005

The epidermis is a physiological barrier to protect organisms against environment. During the aging process, skin tissues undergo various changes including morphological and functional changes. The transcriptional regulation of genes is part of cellular reaction of aging process. In order to examine the changes of gene expression during the aging process, we used the primary cell culture system of human keratinocytes. Since UV radiation is the most important environmental skin aggressor, causing skin cancer and other problems including premature skin aging, we examined the changes of gene expression in human keratinocytes after UV irradiation using oligonucleotide microarray containing over 10,000 genes. We also compared the gene expression patterns of the senescent and UV treated cells. Expression of the variety of genes related to transcription factors, cell cycle regulation, immune response was altered in human keratinocytes. Some of down-regulated genes are represented in both senescent and UV treated cells. The results may provide a new view of gene expression following UVB exposure and aging process in human keratinocytes.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.28 no.1
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• pp.186-201
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• 2002

Fructan(Levan)은 식물체 및 미생물에서 발견되는 탄수화물로 이는 과당(fructose)이 $\beta$-2, 6 결합으로 연결되어 있는 polysaccaride 이다. 본 연구에서는 Fructan을 생성하는 미생물(Zymomonas mobilis)과 10% sucrose(기질), 1-2% 효모 추출물을 주성분으로 하는 배지를 사용하여 30-37$^{\circ}C$, pH 5.0-7.0에서 20-24시간 동안 배양한후 원심분리하여 균체를 제거하고 3배량의 알코올을 가하여 침전, 건조하여 얻은 Fructan의 화장품 원료로서의 가능성을 조사하였다. 보습효과에 있어서는 Hyaluronic acid와 유사하였으며, keratinocyte에 대한 세포증식 효과를 나타내었다. 또한 3-D culture에 의해 구축된 생인공 피부내에 0.05%의 sodium lauryl sulfate (SLS)를 사용하여 피부자극에 의한 초기 염증 반응을 유도한후 0.01mg/m1, 0.05mg/m1의 Fructan을 각각 처리하였을 때, SLS만을 처리한 생인공피부와 비교하여 세포증식효능을 보였고, SLS 자극물질로 유도된 전염증성 조절인자인 interleukin-l$\alpha$(IL-l$\alpha$)의 분비량을 조사 하였을때 0.01mg/ml, 0.05mg/ml의 Fructan을 처리한 생인공피부의 IL-l$\alpha$ 양이 Fructan을 처리하지 않은 것보다 상대적으로 감소하였다. 이러한 결과로 Fructan이 생인공 피부내 피부 세포의 증식효과를 나타낼 뿐만 아니라, 또한 피부자극물질에 의한 염증반응에 대해 자극완화효능이 있음을 알 수 있었다. 섬유아세포 및 동물을 이용한 안전성 시험에서도 독성이 없는 안전한 원료로 평가되었다.

• Development and Reproduction
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• v.12 no.1
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• pp.51-56
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• 2008

To optimize the cell culture conditions of zebrafish embryonic cells, we compared the efficiency of three types of medium, DMEM, K-NAC and D-NAC. In this study, we showed that the cells grown in K-NAC have better plating efficiency than DMEM, especially in the case of low cell seeding density. However, cells grew slower in K-NAC than those in DMEM in confluent cultures. The effect of 0.1% zebrafish embryo extracts was minimal. The presence of 1% trout serum in culture medium significantly increased the growth rate of cells(p<0.05). No difference was found at $2{\sim}3{\times}10^5$ cell seeding density(p<0.05). At $4{\sim}5{\times}10^5$ cell seeding density, cells grew better in DMEM than K-NAC (p<0.1). The results suggest that supplementation of NAC and A2P in Keratinocyte SFM may improves plating efficiency when cells are plated at low population. No difference was found for cell growth in either medium with 5%, 10% or 15% FBS supplemented (p<0.05). Cells culture in D-NAC grew significantly better than those in DMEM(p<0.05). Our results clearly showed that the use of NAC and A2P in the culture medium has a positive effect on cell growth regardless of the amount of FBS added.

• Journal of Life Science
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• v.28 no.10
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• pp.1147-1155
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• 2018

Agaricus blazei mycelial liquid culture extract (ABMLCE) promoted the production of testosterone (TS) in TM-3 mouse Leydig testis cells. Now, we report that ABMLCE containing eritadenine (EA) as a minor constituent (15.3 mg/100 g) reduced $5{\alpha}-reductase$ 2 ($5{\alpha}-R2$) enzyme activity and dihydrotestosterone (DHT) content which are key constituents for the benign prostatic hyperplasia (BPH) inductions. RWPE-1 prostate cells were grown in a Keratinocyte serum-free medium (K-SFM) containing ABMLCE (0~50 ppm), EA (0~10 ppm,), and finasteride (FS $10{\mu}M$: a positive control) in a 24-well plate for 24 hr. Supernatants collected from cell-cultured media were used for the assay of $5{\alpha}-R2$, superoxide dismutase (SOD), catalase (CAT) and cyclooxygenase-2 (COX-2) enzyme activities, and for TS, DHT, tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) and $interleukin-1{\beta}$ ($IL-1{\beta}$) contents by their assay kits. The $5{\alpha}-R2$ activity and DHT content were proportionally reduced (p<0.05) to concentrations of ABMLCE. The SOD and CAT enzyme activities were significantly (p<0.05) elevated concomitant with ABMLCE concentrations, while COX-2, $TNF-{\alpha}$ and $IL-1{\beta}$ showed reverse results (p<0.05). Similarly, the effects of EA were similar to those of ABMLCE. Efficacies of ABMLCE 50 ppm and EA 10 ppm in $5{\alpha}-R2$ and DHT reduction were similar to those of $10{\mu}M$ FS. These results suggest that ABMLCE and EA reduced $5{\alpha}-R2$ and DHT through their anti-inflammatory and anti-oxidative actions. This implies that ABMLCE containing EA could be a beneficial material in the cure of BPH in humans.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.38 no.4
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• pp.311-319
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• 2012

Seborrheic dermatitis (SD) is the skin disease occurred because of Malassezia yeast which grows on the skin and scalp, and this yeast lives on sebum lipid, and their metabolite, free lipid acids are thought to be the main irritant on skin. To find out effective cosmeceutical ingredients to treat SD symptoms, we established novel cell-based in vitro model mimicking SD symptoms. This in vitro model adopted the co-culture system with primary sebocyte & HaCaT keratinocyte. We used M. globosa yeast extract, arachidonic acid, linoleic acid and dihydrotestosterone as SD inducers. In the co-culture system with optimized concentrations for SD-inducing cocktail, the production of IL-8 and sebum lipids increased up to 2-fold, and then we screened with commercial essential oils by monitoring IL-8 as a key inflammatory biomarker. Then we found that Cinnamomum zeylanicum oil, Mentha arvensis oil effectively down-regulated IL-$1{\alpha}$, IL-6, IL-8 cytokines which over-produced by SD-inducing cocktail. Additionally, two essential oils also showed inhibitory effect on sebum lipid synthesis from primary sebocyte and growth inhibitory effect to M. globosa yeast (MICs were lower than 0.0625 %). Our recent results suggest that Cinnamomum zeylanicum oil and Mentha arvensis oil could be effective natural herbal remedies to relieve or protect scalp seborrheic dermatitis.