• The Korean Journal of Physiology and Pharmacology
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• v.9 no.2
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• pp.131-135
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• 2005

Potassium channels in human skin fibroblast have been studied as a possible site of Alzheimer disease pathogenesis. Fibroblasts in Alzheimer disease show alterations in signal transduction pathway such as changes in $Ca^{2+}$ homeostasis and/or $Ca^{2+}-activated$ kinases, phosphatidylinositol cascade, protein kinase C activity, cAMP levels and absence of specific $K^+$ channel. However, little is known so far about electrophysiological and pharmacological characteristics of large-conductance $Ca^{2+}$-activated $K^+$ ($BK_{Ca}$) channel in human fibroblast (CRL-1474). In the present study, we found Iberiotoxin- and TEA-sensitive outward rectifying oscillatory current with whole-cell recordings. Single channel analysis showed large conductance $K^{+}$ channels (106 pS of chord conductance at +40 mV in physiological $K^+$ gradient). The 106 pS channels were activated by membrane potential and $[Ca^{2+}]_i$, consistent with the known properties of $BK_{Ca}$ channels. $BK_{Ca}$ channels in CRL-1474 were positively regulated by adenylate cyclase activator ($10{\mu}M$ forskolin), 8-Br-cyclic AMP ($300{\mu}M$) or 8-Br-cyclic GMP ($300{\mu}M$). These results suggest that human skin fibroblasts (CR-1474) have typical $BK_{Ca}$ channel and this channel could be modulated by c-AMP and c-GMP. The electrophysiological characteristics of fibroblasts might be used as the diagnostic clues for Alzheimer disease.

• Journal of Life Science
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• v.21 no.9
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• pp.1266-1273
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• 2011

Saponin is a main component of ginseng widely known as an oriental traditional medicinal ingredient. A variety of biological effects of saponin has been reported, but its action related to skin regeneration has remained unclear so far. In this study, the effect of saponin on matrix metalloproteinase as well as its antioxidant effect in cell free system was examined in human dermal fibroblasts. First of all, as a result of investigating the effect of saponin on cell viability using MTT assay, it was shown to increase cell viability below 10 ${\mu}g$/ml, but it also showed cytotoxicity above 25 ${\mu}g$/ml. The antioxidant effect of saponin was exerted by inhibition of $H_2O_2$ in addition to reducing power above 1 ${\mu}g$/ml. In particular, saponin showed a protective effect on DNA oxidation. Furthermore, it was observed that saponin activates MMP-2 and increases MMP-1 activity in gelatin and casein zymography analyses, respectively, indicating that saponin could have potential a therapeutic agent for anti-aging and skin regeneration.

• Archives of Plastic Surgery
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• v.35 no.2
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• pp.115-120
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• 2008

Purpose: Oncostatin M(OSM) is a multifunctional cytokine that belongs to the interleukin(IL)-6 family. Although there have been a number of studies that focused on the role and mechanism of OSM in various organs and tissues, there are few reports on its effect on wound healing. The final purpose of this project is to evaluate the effect of OSM on wound healing. This pilot study was designed to investigate the effect of OSM on proliferation and matrix synthesis of human dermal fibroblasts, which are the major components of the wound healing. Methods: Excess skin that was obtained from patients who underwent skin grafts, was used for this study. From this material, fibroblasts were isolated and cultured. The cultured fibroblasts were treated with one of four concentrations of OSM. The OSM concentrations used were 0, 50, 100, and 200 ng/ml, respectively. After the OSM treatment, cell proliferation was determined by the MTT assay, collagen synthesis by the C1CP method, GAG levels by the Blyscan Dye method. The parameter levels of each group were compared. Results: OSM treatment increased all the components tested in the study. In particular, cell proliferation, GAG synthesis demonstrated statistically significant increases(p<0.05 in the Mann-Whitney U-test). The highest increase in all the components was obtained at a 100 ng/ml concentration of OSM.Conclusion: The results of the present study indicate that OSM stimulates proliferation and matrix synthesis of human dermal fibroblast and the optimal concentration for wound healing is 100 ng/mL.

• Journal of Ginseng Research
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• v.44 no.5
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• pp.738-746
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• 2020

Background: Red ginseng contains components, including microelements, vitamins, essential oils, and fatty acids, that can be used in skincare to delay the aging process. We investigated the effects of red ginseng treatment on skin elasticity by assessing cellular stiffness and measuring collagen protein synthesis. Methods: Human dermal fibroblasts were treated with red ginseng, and the resulting changes in stiffness were investigated using atomic force microscopy. Cytoskeletal changes and mRNA expression of biomarkers of aging, including that of procollagens I and VII, elastin, and fibrillin-1, were investigated. Collagen in a human skin equivalent treated with red ginseng was visualized via hematoxylin and eosin staining, scanning electron microscopy, and atomic force microscopy. Results and conclusion: The stiffness of fibroblasts was significantly reduced by treatment with red ginseng concentrations of ≥ 0.8 mg/mL. The ratio of F-actin to G-actin decreased after treatment, which corresponded to a change in fibroblast stiffness. The storage modulus (G') and loss modulus (G'') of the skin equivalent were both lowered by red ginseng treatment. This result indicates that the viscoelasticity of the skin equivalent can be restored by red ginseng treatment.

• YAKHAK HOEJI
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• v.54 no.2
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• pp.112-121
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• 2010

The cytotoxic activity of 33,4,5-trihydroxybenzoic acid methylester and related compounds on the growth of normal cell lines, human skin melanoma cells and human oral epithelioid cell line were evaluated by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and 2,3-bis-[2-methoxy-4-nitro-5-sulfo-phenyl]-2-H-tetrazolium-5-caboxanilide (XTT) methods. 3,4,5-Trihydroxybenzoic acid methylester decreased the cell viability of human skin melanoma cells and human oral epithelioid cells shown by the MTT method and the cell adhesion activity of human skin melanoma cells and human oral epithelioid cells shown by the XTT method. In light microscopy, 100 ${\mu}M$ 3,4,5-trihydroxybenzoic acid methylester showed the highest cytotoxic activity. These results suggest that 3,4,5-trihydroxybenzoic acid methylester has a potential anticancer activity.

• 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.

• Korean Journal of Pharmacognosy
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• v.54 no.1
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• pp.1-8
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• 2023

The skin is an important barrier that protects the body from harmful environments such as UV rays. When the skin is repeatedly stimulated, such as UV rays, ROS and pro-inflammatory cytokines are overproduced. As a result, the proteins and nucleic acids that make up the skin are damaged, and aging occurs. Esculetin is known to have anti-inflammatory, antioxidant and UV-induced MMP-1 inhibitory effects. However, the inhibitory effect of MMP-1 on TNF-α-induced fibroblasts is not known. Therefore, in this study, the MMP-1 inhibitory effect of esculetin was confirmed in TNF-α-induced fibroblasts. As a result of confirming the cytotoxicity of esculetin in Hs68 cells by MTT assay, there was no significant toxicity up to 200 µM. As a result of real-time PCR and ELISA, it was confirmed that esculetin inhibited the expression of MMP-1. Esculetin did not inhibit MAPK (ERK, JNK, p38) phosphorylation, but inhibited phosphorylation of the mTOR-p70S6k signaling pathway. In addition, it was confirmed that the phosphorylation of the transcription factor NF-κB was inhibited. These results suggest that esculetin has potential as an anti-aging material.

• 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.

• BMB Reports
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• v.53 no.10
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• pp.539-544
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• 2020

Skin aging appears to be the result of overlapping intrinsic (including genetic and hormonal factors) and extrinsic (external environment including chronic light exposure, chemicals, and toxins) processes. These factors cause decreases in the synthesis of collagen type I and elastin in fibroblasts and increases in the melanin in melanocytes. Collagen Type I is the most abundant type of collagen and is a major structural protein in human body tissues. In previous studies, many products containing collagen derived from land and marine animals as well as other sources have been used for a wide range of purposes in cosmetics and food. However, to our knowledge, the effects of human collagen-derived peptides on improvements in skin condition have not been investigated. Here we isolate and identify the domain of a human COL1A2-derived protein which promotes fibroblast cell proliferation and collagen type I synthesis. This human COL 1A2-derived peptide enhances wound healing and elastin production. Finally, the human collagen alpha-2 type I-derived peptide (SMM) ameliorates collagen type I synthesis, cell proliferation, cell migration, and elastin synthesis, supporting a significant anti-wrinkle effect. Collectively, these results demonstrate that human collagen alpha-2 type I-derived peptides is practically accessible in both cosmetics and food, with the goal of improving skin condition.

• Journal of the Korean Society of Food Culture
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• v.35 no.5
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• pp.478-482
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• 2020

We evaluated the protective effects of cricket methanol extract (CME) on ultra-violet B (UVB)-induced photoaging in human skin fibroblasts. The fibroblast cells were treated with 10, 50, and 100 ㎍/mL of CME for 24 h, and then exposed to UVB (30 mJ/㎠). CME showed a dose-dependent cytoprotective effect without any observable cytotoxicity. CME reduced UVB-induced production of reactive oxygen species (ROS) by 34.4, 34.9, 40.6% at concentrations of 10, 50, 100 ㎍/mL respectively. CME inhibited the release of matrix metalloproteinase (MMP) 1 and 3. Furthermore, CME also reduced UVB-induced collagen degradation in the fibroblast cells. Taken together, our data suggests that CME has a significant protective effect on UVB-induced photoaging of the skin. This benefit occurs through multiple mechanisms. The results also suggest a potential role for CME as an ingredient in anti-photoaging cosmetic products in the future.