• Journal of the Society of Cosmetic Scientists of Korea
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• v.23 no.3
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• pp.97-100
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• 1997

These days, the raw materials that have the cell recovering effect are used commonly in cosmetics. In this study, six materials were rested for the characteristics of recovering effect both on vivo and in vitro. Tested raw materials were Soypol, 3-APPA, Apple extract, Polygonatum japonicum extract, Scutellarkd baicalensis extract, Aloe extract. Among these materials, Soypol and 3-APPA were synthesized and others were made by extraction at the Pacific R&D Center. Human forearm skin and cultured skin cell were damaged by sodium lauryl sulfare and then raw materials were applied for open treatment on SLS damaged human skin or cells. The recovering effects of raw materials in vivo were evaluated by measuring transepidermal water loss, skin hydration and erythema and in vitro effects of proliferationg cells were assessed by neutral red uptake assay. In the in vivo study, only the evaluation by TEWL showed correlation with the visual score. Our of six materials, 3-APPA had the most positive effect in both in vivo and in vitro studies and the correlation was r=0.8286 (p=0.042).

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.2 s.33
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• pp.47-67
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• 2007

Objective : As a result of increasing amount of ultraviolet ray, skin problems including sunburn, rapid skin aging, melanoma, and even skin cancer continue to rise. In the present study, the effect of oriental herbal extract, Daehwanggo(大黃膏,DH) and Daehwanggogasnagbakpi(大黃膏加桑白皮,DS), as external application, on the skin damage, was investigated. Methods : 30 mice were equally distributed into 3 groups : control, UVB-control and UVB-irradiated and DS-treated group. Also mouse melanoma cell lines were cultured. Tyrosinase inhibition was measured to analyze the UN-protection effect. Melanogenesis in the UV-irradiated melanoma cell lines was compared in DS-treated cell line and control cell line. Sample skin from the ear tissue of the 3 groups were analyzed to observe the inflammatory response, T cell differentiation, apoptosis of keratinocytes. Results : The tyrosinase was more significantly inhibited in the DS group compared to DH group. Antioxidative effects was more prominent in DS group when superoxide dismutase was measured. Both the DS- and DH-treated cell lines showed significantly reduced melanogenesis. The reduction of external skin damage including erythematous papule, eczema, keratinocyte, pyopoiesis was observed in the DS- and DH-treated sample cells. In terms of the effect on the skin damage, sunburn cell, activated skin mast cells, secretion of IL-12, manifestation of HSP70, hyperplasia of epithelial cells, MMP-9 and destruction of the collagen were all significantly improved in the DS-treated sample cells. Melanin cells and the apoptosis in the melanoma cell line were decreased. Conclusion : DH and DS were traditionally applied externally for the scald in the oriental medicine. The present study elucidated the possibility of herbal extracts to be used as ultraviolet protectives. Further investigations are needed to assure the clinical application.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.25 no.4 s.34
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• pp.57-63
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• 1999

Pericarpium castaneae extracts have variously potent activities, such as anti-oxidative activity and free radical scavenging activity. in vivo and in vitro studies both indicate that pericarpium castaneae extracts acts as a free radical scavenger($IC_{50}:7.6{\mu}g/ml$) stronger than gallic acid($IC_{50}:12.5{\mu}g/ml$) and ellagic acid($IC_{50}:15{\mu}g/ml$) which could prevent cutaneous UV damages and skin aging. The extracts showed a good effect as a anti-oxidant($IC_{50}:50{\mu}g/ml$). It was shown that the appearance of wrinkle in human skin was reduced by topical application of pericarpium castaneae extracts. And the treatment of human skin with the extracts increased the elasticity and moisture of the skin. We investigated the effect of the pericarpium castaneae extracts on production of extracellular matrix using cultured A431 fibroblast cells. The results indicated that the extracts had no detectable effect on collagen synthesis. But synthesis of cell adhesion protein was increased by the extracts. The results suggest that increase of cell adhesion protein synthesis by pericarpium castaneae extracts has closely related to reduction of wrinkle in skin.

• Proceedings of the SCSK Conference
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• 1999.10a
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• pp.57-64
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• 1999

Pericarpium castaneae extracts have variously potent activities, such as anti-oxidative activity and free radical scavenging activity. in vivo and in vivo studies both indicate that pericarpium castaneae extracts acts as a flee radical scavenger ($IC_{50}$/: 7.6$\mu\textrm{g}$/ml) stronger than gallic acid($IC_{50}$/: 12.5$\mu\textrm{g}$/ml) and ellagic acid($IC_{50}$/: 15$\mu\textrm{g}$/ml) which could prevent cutaneous UV damages and skin aging. The extracts showed a good effect as a anti-oxidant ($IC_{50}$/: 50$\mu\textrm{g}$/ml). It was shown that the appearance of wrinkle in human skin was reduced by topical application of pericarpium castaneae extracts. And the treatment of human skin with the extracts increased the elasticity and moisture of the skin. We investigated the effect of tile pericarpium castaneae extracts on production of extracellular matrix using cultured A431 fibroblast cells. The results indicated that the extracts had no detectable effect on collagen synthesis, But synthesis of cell adhesion protein was increased by the extracts. The results suggest that increase of cell adhesion protein synthesis by pericarpium castaneae extracts has closely related to reduction of wrinkle in skin.

• Biomolecules & Therapeutics
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• v.22 no.4
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• pp.328-333
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• 2014

Vitiligo is a pigmentary disorder induced by a loss of melanocytes. In addition to replacement of pure melanocytes, cocultures of melanocytes with keratinocytes have been used to improve the repigmentation outcome in vitiligo treatment. We previously identified by in vitro studies, that adipose-derived stem cells (ADSCs) could be a potential substitute for keratinocytes in cocultures with melanocytes. In this study, the efficacy of pigmentation including durability of grafted melanocytes and short-term safety was examined in the nude mouse and Sprague-Dawley rat after grafting of primary cultured human melanocytes, with or without different ratios of primary cultured human ADSCs. Simultaneous grafting of melanocytes and ADSCs, which were separately cultured and mixed on grafting at the ratios of 1:1, 1:2, or 1:3, showed better efficacy than that of pure melanocytes. Grafting of melanocytes cocultured with ADSCs resulted in a similar outcome as the grafting of cell mixtures. Skin pigmentation by melanocytes : ADSCs at the ratios of 1:1 and 1:2 was better than at 1:3. No significant difference was observed between the 1-week and 2-week durations in coculturing. Time-course microscopic examination showed that the grafted melanocytes remained a little longer than 6-week post-grafting. No inflammatory cell infiltration was observed in the grafted skin and no melanocytes were detectable in other organs. Collectively, grafting of melanocytes and ADSCs was equally safe and more effective than grafting of melanocytes alone. Despite the absence of significant differences in efficacy between the group of 1:1 and that of 1:2 ratio, 1:2 ratio for 1-week coculturing may be better for clinical use from the cost-benefit viewpoint.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.69-69
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• 2001

The cell cycle phase in which donor nuclei exist prior to nuclear transfer is an important factor governing developmental rates of reconstituted embryos. It was suggested that quiescent G0 and cycling G1 cells could support normal development of reconstituted embryos. In a quest of optimized donor nuclei treatment prior to nuclear transfer, this study was undertaken to examine the cell cycle characteristics of bovine fetal and adult somatic cells when cultured under a variety of culture treatments and the cell cycle change with the lapse of time after trypsinization. This was archived by measuring the DNA content of cells using flow cytometry, Cultured fetal fibroblast cells, adult skin and muscle cells, and cumulus cells were divided by 3 culture treatments; 1) grown to 60-70% confluency (cycling), 2) serum starved culture, 3) culture to confluency. Trypsinized cells were fixed by 70% ethanol and stained with propidium iodide. For one experiment, trypsinized cells were resuspended in DMEM＋10% FBS and incubated for 1.5, 3 and 6 h with occasional shaking before ethanol fixation. Cell cycle phases were determined by flow cytometry enabling calculation of percentages of G0＋G1, S and G2＋M. The majority of cells were in G0＋Gl stage regardless of origin of cells. Cultures that were serum starved or cultured to confluency contained significantly (P<0.05) higher percentages of cells in G0＋G1 (89.5-95.4%). For every cell lines and culture treatments, percentages of cells in existing in G0＋G1 increased with decreasing of the cell size from large to small. In the serum starved and confluency groups, about 98% of small cells were in G0＋G1 Serum starved culture contained higher percentages of small-sized cells (38.5-66.9%) than cycling and confluent cultures regardless of cell lines (P<0.05). After trypsinization of fetal fibroblast and adult skin cells that were serum starved and cultured to confluency, the percentages of cells in G0＋G1 significantly increased by incubation for 1.5(95.7-99.5%) and 3.0 h (95.9-98.6%). The results suggest that the efficient synchronization of bovine somatic cells in G0＋G1 for nuclear transfer can be established by incubation for a limited time period after trypsinization of serum starved or confluent cells.

• Journal of Animal Reproduction and Biotechnology
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• v.36 no.3
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• pp.137-144
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• 2021

In this study, to analyze whether artificial regulation of apoptosis in the development of somatic cells can affect the stable growth and development of cells, 20 alpha-hydroxysteroid dehydrogenase (20α-HSD) and rapamycin were treated to induce apoptosis and autophagy in the both skin and muscle cells. Respectively, and 3-methyladenine was supplemented to inhibit cell death. Our results show that stimulation with rapamycin activated autophagy in both types of cells, but increased apoptosis more than autophagy in the case of skin cells. These results indicate that there was a difference in the expression of survival factors and cell development depending on the type of cell. In particular, in the expression of autophagy-related gene (MAP1LC3A) was higher than that of Casp-3, an apoptosis factor. Furthermore, cell development was the highest in all cell groups cultured by artificially inducing autophagy, however the lowest in the apoptosis-inhibited group. Especially, the noteworthy result of this study was that when apoptosis was induced using 20α-HSD, it was possible to induce apoptosis in both skin and muscle cells. Therefore, the main point of this study is that apoptosis induced during cell culture plays a pivotal role in cell remodeling.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.39 no.2
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• pp.63-70
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• 2013

Objectives: The purpose of this study was to investigate the wound healing effect of primary cultured oral mucosal keratinocytes (OMKs) and to assess their roles in skin wounds. Materials and Methods: OMK labeled with BromodeoxyUridine were scattered onto $1.5{\times}1.5$ cm skin defects of adult female nude mice (OMK group, n=15). For the control, culture media were placed on the wound (control group, n=15). Mice in both groups were sacrificed at three days (n=5), one week (n=5), and two weeks (n=5), and histomorphometric and immunoblot analyses with keratinocyte growth factor (KGF), interleukin (IL)-6, and IL-$1{\alpha}$ antibody were performed for the biopsied wound specimen. To verify the effect of the cytokine, rhIL-$1{\alpha}$ was applied instead of OMK transplantation, and the OMK and control groups were compared with regard to re-epithelialization. Results: Histomorphometric analyses demonstrated faster re-epithelialization in the graft group than in the control group at the third day, first week, and second week. Newly forming epithelium showed maintenance of the histological character of the skin epithelium. The graft group showed superior expression of KGF, IL-6, and IL-$1{\alpha}$ protein, compared with the control group. Similar faster re-epithelialization was observed after treatment with rhIL-$1{\alpha}$ instead of OMK transplantation. Conclusion: We successfully confirmed that the graft of primary cultured OMKs promoted regeneration of skin defects. The mechanism of accelerated wound healing by primary cultured OMKs was attributed to inducement of cytokine expression as required for re-epithelialization.

• Journal of Haehwa Medicine
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• v.28 no.2
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• pp.41-47
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• 2019

Objectives : Atopic dermatitis is an irritable skin disease accompanying rash and itching leading to impaired skin barrier. ATO-ALL is an ethanol extract of natural products comprising 12 herbs and effective on atopic dermatitis. In this study, we aimed to propose that the effect of ATO-ALL on skin regeneration in human keratinocyte cell line, HaCaT cells. Methods : To evaluate the skin regenerating effects of ATO-ALL, scratch wound healing assay, bromodeoxyuridine (BrdU) assay, and propidum iodide (PI) assay were performed using cultured HaCaT cell line. Result : Scratch wound healing assay showed that ATO-ALL was able to enhance the gap filling activity more than 2-fold at 7 ppm concentration compared with control group. BrdU assay demonstrated that ATO-ALL treatment increased the de novo cell proliferation in a dose-dependent manner. Finally, PI assay indicated that the cell cycle of HaCaT cells was modulated by ATO-ALL treatment. Conclusions : These results suggested that ATO-ALL may have skin regenerating effects by increasing cell proliferation via cell cycle regulation. Taken together, ATO-ALL is supposed to have a potential on regeneration of damaged skin or functional disease including atopic dermatitis.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.302-305
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• 2003

It is difficult to obtain sufficient healthy skin for coverage of a wide area of skin wound. In the skin, an additional population of living epithelial cells is located in the outer root sheath (ORS) of hair $follicles.^{1),2)}$ ORS cells should be a good source of epithelium because they are easily obtainable and patients do not have to suffer from scar formation at donor sites. We modified ordinary primary culture technique for the purpose of solving such problem that epithelial cells have a low propagation and easy aging during culture periods. First of all, we improved primary cultivation methods. In the ordinary primary culture, average yield of human ORS cells was $2\;{\times}\;10^3$ cells/follicle by direct incubation with trypsin (0.1%)/EDTA (0.02%) solution for 15 min at $37^{\circ}C$ but we could obtain about $6.5\;{\times}\;10^3$ cells/follicle by two step enzyme digestion method with dispase (1.2 U/ml) and trypsin (0.1%)/EDTA (0.02%) solution. So we could achieve three times higher primary cultured ORS cell yield. Secondly, we could obtain total $2\;{\times}\;10^7$ cells in serum free medium and even more total $6\;{\times}\;10^7$ cells in modified E-medium with mitomycin C-treated feeder cells during 17 days. Using the cultured ORS cells, and we could make bioartificial skin equivalent in vitro and concluded that ORS cells were progenitor cells for skin epithelial cell.