• Korea Journal of Cosmetic Science
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• v.1 no.1
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• pp.1-9
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• 2019

The identification of compounds with anti-senescence activity in cell culture system is a first step in aging research. Given that pyruvate can be used energy source by conversion to acetyl-CoA in mitochondria, and protects cultured cell from various stress-induced cell damage and cell death, synthetic media (e.g., DMEM) often includes 1 mM pyruvate, which is very higher than the pyruvate concentration in human blood (approximately 30 ��M). However, the use of medium containing high concentration of pyruvate is not suitable for screening anti-senescence compounds, because pyruvate also protects against the cellular senescence of primary human dermal fibroblasts (NHDFs) through NAD+ generated during conversion to lactate. In this study, four extracts, i.e., Sprouted seed and fruit complex, Poncirus trifoliata fruit extract, Jaum balancing complex, and Prunus mume extract were used for evaluation of different anti-senescence effect in the absence or presence of 0.1 mM pyruvate, similar to the physiological pyruvate concentration. The senescence in NHDFs cultured with DMEM in the presence of 0.1 mM pyruvate (approximately the physiological concentration in human blood) is accelerated, as observed in pyruvate deprivation conditions. The cytotoxicity of the Poncirus trifoliata fruit extract was protected by pyruvate, and Jaum balancing complex and Prunus mume extract had anti-senescence activity in the presence of 0.1 mM pyruvate, but not in the absence of pyruvate. Given that pyruvate is a powerful protector against both cytotoxicity and cellular senescence, the screening of candidate agents for anti-senescence in high pyruvate conditions using an in vitro cell culture system is not valid. Therefore, we recommend the use of a low concentration of pyruvate to evaluate the anti-senescence effects of candidates, which is more similar to in vivo aging conditions than excessive stress-induced senescence models, to exclude the effect of excessive pyruvate in vitro.

• Archives of Pharmacal Research
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• v.26 no.10
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• pp.855-860
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• 2003

The effect of silver sulfadiazine (SSD) on the proliferation of human dermal fibroblast (HDF) was studied to determine the impact of the drug on the wound healing process and dermal mechanical strength. Human dermal fibroblasts were cultured to 80% confluency using DMEM with 10% FBS and viability of the cell was estimated using neutral red assay. In addition, the $2^{nd}$ degree burn wound was prepared on the anterior part of rabbit ear skin and dressings containing SSD were applied for 96 h. Presence of inflammatory cells and degree of re-epithelialization were investigated in the wound. After 15 day of the induction of burn wounds, the treated area was excised and dermal mechanical strength was quantitatively measured with a constant speed tensiometer. SSD was found to be highly cyto-toxic in cultured HDF cells. The topical application of SSD (2%) could control the infection as evidenced by the lack of accumulation of inflammatory cells in histological evaluation. Therefore, these observations suggested that the impairment of dermal regeneration and decreased mechanical strength of dermal tissue was resulted from the cyto-toxic effect of SSD on dermal cells. Since the decreased mechanical strength may lead to reduction in resilience, toughness and maximum extension of the tissue, the identification of optimum dose for SSD that limits infection while minimizes the cyto-toxic effect may be clinically relevant.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.42 no.1
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• pp.87-94
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• 2016

Skin damage is mainly caused by environmental factors such as ultraviolet light, heat, and smoking. It is known that reactive oxygen species production is commonly involved in the pathogenesis of skin damage induced by these factors, causing skin aging. Pyrus pyrifolia Nakai continues to be a popular and highly consumed fruit in many countries with known beneficial effects including antitumor, antioxidative, and anti-inflammatory effects. However, there is no evidence of a therapeutic effect of Pyrus pyrifolia extract (PPE) against skin aging via inhibition of mitochondria-mediated apoptosis. In this study, we investigated PPE protective effect against photoaging induced by UVB ($50mJ/cm^2$) in HS68 human dermal fibroblasts. Lactate dehydrogenase assay showed that PPE significantly protected HS68 cells against UVB-induced damage in a dose-dependent manner. Other assays using DCF-DA demonstrated that PPE protected HS68 cells by regulating reactive oxygen species production. PPE also regulated mitochondrial dysfunction and mitochondrial membrane potential induced by UVB, and inhibited UVB-induced caspase-3 activity. These results indicate that PPE protects human dermal fibroblasts from UVB-induced damage by regulating the oxidative defense system.

• Journal of Life Science
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• v.21 no.11
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• pp.1501-1510
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• 2011

In recent years novel potential pharmocological candidates have been looked for in animal, seaweed, sponge, fungi and marine bacteria resources. In this study, matrix metalloproteinases (MMPs) that play an important role in metastasis, arthritis, chronic inflammation and wrinkle formation were used as target enzymes to screen therapeutic agents. The inhibitory effects of several marine algae including green algae (5 species), red algae (18 species) and brown algae (4 species) methanolic extracts on MMPs were investigated in human dermal fibroblasts and human fibrosarcoma cell line (HT1080 cells) using gelatin zymography. In human dermal fibroblasts, the inhibition of MMP-2 was observed in Laurencia okamurae, Polysiphonia japonica, Grateloupia lanceolate and Sinkoraena lancifolia of red algae. In contrast, MMP-2 activation was enhanced in Enteromorpha compressa and E. linza of green algae, and Peltaronia bighamiae and Sargassum thunbergii of brown algae. In human fibrosarcoma cells, MMP-9 activation was decreased in the presence of S. thunbergii of brown algae, Polysiphonia japonica in red algae and E. compressa and E. linza of green algae. The interesting finding is that E. compressa and E. linza of green algae, and S. thunbergii of brown algae exhibited a positive effect on MMP-2 in normal cells, but a negative effect on MMP-9 in cancer cell lines. These results suggest that E. compressa and E. linza of green algae, and S. thunbergii of brown algae contain potential therapeutic ingredients for cancer treatment.

• Archives of Plastic Surgery
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• v.32 no.3
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• pp.343-346
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• 2005

It has been established that a graft of fibroblasts is able to improve wound healing. However, there has been no research on the effect of a graft of bone marrow stromal cells on wound healing. The wound healing process requires cell proliferation and production of extracellular matrix and various growth factors. The purpose of this study was to compare the abilities of human fibroblasts and bone marrow stromal cells, which contains mesenchymal stem cells, to proliferate and to produce collagen. Human bone marrow stromal cells and fibroblasts were isolated from bone marrow and dermis of the same patients and grown in culture respectively. Cell proliferation and production of type I collagen by human bone marrow stromal cells and dermal fibroblasts were examined by MTT method and by ELISA of cell culture media on day 1, 3, and 5 days post-incubating. The human bone marrow stromal cells showed 11-17% higher cell proliferation than fibroblasts at each time interval. The levels of type I collagen in the human bone marrow stromal cell group was also significantly higher than those in the fibroblast group. The results indicate that the grafts of human bone marrow stromal cells can show more promising effect than that of fibroblasts for healing of chronic wounds.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.32 no.2 s.57
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• pp.75-79
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• 2006

In this study. we evaluated anti-aging activity of medical plants that protect the skin cell damage induced by UV irradiation. We have investigated diverse biological activities of Selaginella tamariscina as an anti-aging ingredient of cosmetics. S. tamariscina was found to show scavenging activities of radicals and reactive oxygen species (ROS) with the $IC_{50}$ values of $65.1{\mu}g/mL$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $40.9 {\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. For testing intracellular ROS scavenging activity, the cultured human dermal fibroblasts were analyzed by increase in dichlorofluorescein (DCF) fluorescence upon exposure to UVB $20 mJ/cm^2$ after treatment of S. tamariscina. UVA-induced MMP-1 protein and mRNA expression in human dermal fibroblasts were reduced in a dose-dependent manner by S. tamariscina. Moreover, S. tamariscina inhibited MMP-2 (gelatinase) activity in UVA-irradiated human dermal fibroblasts assayed by zymography and semi-quantitative RT-PCR. Taken together, these results suggest that S. tamariscina may act as an anti-aging agent by Increasing collagen and preventing the skin cell damage induced by UV irradiation, and imply that S. tamariscina nay be useful as a new ingredient for anti-aging cosmetics.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.301-306
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• 2011

Fibroblasts of large animals are easy to isolate and to maintain in vitro culture. Thus, these cells are extensively applied to donor cell for somatic cell nuclear transfer, and to substrate cells to generate induced pluripotent stem cells after transfection of requited genes to be essentially required for direct reprogramming. However, limited mitotic activity of fibroblasts to differentiate along a terminal lineage becomes restrictive for their versatile application. Recently, commercial culture medium and systems developed for primary cells are provided by manufactures. In this study, we examined whether one of the systems developed for primary fibroblasts of human are effective on porcine ear skin fibroblasts. To this end, we performed proliferation assay after five days culture in vitro of porcine fibroblasts in medium DMEM, which is generally used for fibroblasts culture, and medium M106 for human dermal fibroblasts, supplemented with various concentrations of FBS and LSGS contained mainly growth factors, respectively. Consequence was that presence of 15% FBS and 0.1 ${\times}$ concentrations of LSGS in DMEM showed most active proliferation of porcine fibroblasts.

• Journal of Periodontal and Implant Science
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• v.36 no.3
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• pp.757-765
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• 2006

Human periodontal ligament fibroblasts (hPDLF) are very important for curing the periodontal tissue because they can be differentiated into various cells. A tissue engineering approach using a cell-scaffold is essential for comprehending today's periodontal tissue regeneration procedure. This study examined the possibility of using an acellular dermal matrix as a scaffold for human periodontalligament fibroblast (hPDLF). The hPDLF was isolated from the middle third of the root of periodontally healthy teeth extracted for orthodontic reasons. The cells were cultured in a medium containing Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at $37^{\circ}C$ in humidified air with 5% $CO_2$. The acellular dermal matrix(ADM) was provided by the US tissue banks(USA). Second passage cells were used in this study. The hPDLF cells were cultured with the acellular dermal matrix for 2 days, and the dermal matrix cultured by the hPDLF was transferred to a new petri dish and used as the experimental group. The control group was cultured without the acellular dermal matrix, The control and experimental cells were cultured for six weeks. The hPDLF cultured on the acellular dermal matrix was observed by Transmission Electron microscopy (TEM). Electron micrography shows that the hPDLF was proliferated on the acellular dermal matrix. This study suggests that the acellular dermal matrix can be used as a scaffold for hPDLF.

• 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).

• 대한치주과학회:학술대회논문집
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• 2007.11a
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• pp.154-155
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• 2007