• Proceedings of the SCSK Conference
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• 2003.09a
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• pp.804-819
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• 2003

It has been known that adenine is a very important material in living cells. Because, adenine is a member of nucleotide base, so it takes part in DNA, RNA and ATP synthesis. There are many reports that adenine participated in ingredients, especially DNA, RNA, NADH and ATP, affect on the cell. As well adenosine, conjugated adenine to glycoside, was known to anti-wrinkle compound. But there is no report whether adenine shows a good effect on the skin, especially anti-wrinkle. So, in this study, we tested whether adenine affects cell proliferation, collagen synthesis, collagenase synthesis inhibition in human dermal fibroblasts. In addition, we performed clinical study with adenine cream. Cell proliferation effect was tested by MTT assay. Collagen and collagenase synthesis were measured by Immunoassay with ELISA kit. Clinical study was performed by IECK according to KFDA Functional Cosmetic method. The results of cell proliferation show that 10$^{-6}$ ~10$^{-8}$ ％ of adenine increases cell proliferation about 50 ％ compare with non-treated control. At 10$^{-7}$ ~10$^{-10}$ ％, adenine increases type I collagen synthesis about 50％, decreases type I collagenase about 22％ compare with non-treated control. The results of clinical study show that 0.05％ adenine treated group reduces wrinkle significantly compare with placebo treated group. Therefore adenine may be a new anti-wrinkle candidate, through increases cell proliferation and collagen synthesis dramatically. And it decreases collagenase synthesis. So adenine could be used as a new anti-wrinkle agent.

• Journal of Microbiology and Biotechnology
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• v.33 no.9
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• pp.1250-1256
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• 2023

Herein, different extracts of Scenedesmus deserticola JD052, a green microalga, were evaluated in vitro as a potential anti-aging bioagent. Although post-treatment of microalgal culture with either UV irradiation or high light illumination did not lead to a substantial difference in the effectiveness of microalgal extracts as a potential anti-UV agent, the results indicated the presence of a highly potent compound in ethyl acetate extract with more than 20% increase in the cellular viability of normal human dermal fibroblasts (nHDFs) compared with the negative control amended with DMSO. The subsequent fractionation of the ethyl acetate extract led to two bioactive fractions with high anti-UV property; one of the fractions was further separated down to a single compound. While electrospray ionization mass spectrometry (ESI-MS) and nuclear magnetic resonance (NMR) spectroscopy analysis identified this single compound as loliolide, its identification has been rarely reported in microalgae previously, prompting thorough systematic investigations into this novel compound for the nascent microalgal industry.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.47 no.2
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• pp.147-153
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• 2021

Collagen hydrolysate (CH) is known to prevent skin aging by stimulating skin dermal fibroblasts to promote synthesis of extracellular matrix such as collagen and elastin. Recently, among the various factors that cause skin aging, advanced glycation end products (AGEs) have received particular attention. However, the effect of CH on AGE accumulation has not been studied. Since CH could affect AGE accumulation by promoting production of skin structural proteins, clinical trial was performed using low molecule collagen peptide (LMCP), which were CH containing 25% tripeptide and 4% Gly-Pro-Hyp. Skin autofluorescence (SAF) values were measured using an AGE reader to evaluate accumulation of AGE in skin. As a result of applying 0.5% and 1.0% LMCP solutions to the subject's forearm for 8 weeks, the SAF value at the test site significantly decreased compared to the control site. Additionally, in vitro test was performed using CCD-986sk to evaluate the promotion of collagen synthesis in skin fibroblasts by LMCP. As a result, 800 ㎍/mL of LMCP significantly increased synthesis of human pro-collagen Iα1 (COL1A1) in CCD-986sk. Through this study, we have confirmed that tropical LMCP applications can promote collagen synthesis to help anti-glycation effects, suggesting that LMCP has potential as an anti-aging cosmetic material.

• Korean Journal of Food Science and Technology
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• v.52 no.4
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• pp.369-376
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• 2020

The purpose of this study was to evaluate the antioxidant components and activities of HR02/04(8:2)-W, a mixture of S. quelpaertensis Nakai and F. erecta var. sieboldii. We investigated the p-coumaric acid, total flavonoid, and total phenol contents. To evaluate the antioxidant efficacy, we measured the 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzthiazoline6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity, FRAP activity, reducing power, and ORAC value. We observed the protective effect of hydrogen peroxide against cell damage in human dermal fibroblasts. As a result of the experiment, the p-coumaric acid, total flavonoid, and total phenol contents were 75.62±1.56 mg/100 g, 21.57±0.84 mg rutin equivalent (RE)/g, and 21.25±1.31 mg gallic acid equivalent (GAE)/g, respectively. In the experiments on antioxidant activity, HR02/04(8:2)-W was found to have significantly increased antioxidant activity. In the human dermal fibroblasts, the HR02/04(8:2)-W treated groups could effectively protect cells against oxidative damage. In this study, we confirmed that HR02/04(8:2)-W is a material with effective physiological antioxidant activity.

• Archives of Plastic Surgery
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• v.35 no.5
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• pp.501-506
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• 2008

Purpose: In order to overcome the limitations of the conventional cryopreserved fibroblast or keratinocyte allograft method used in the treatment of diabetic foot ulcers, we reported a pilot study in 2004 demonstrating promising results of a fresh fibroblast allograft method in eight patients. However, the number of cases was insufficient for full evaluation and the follow-up duration was not long enough to determine the efficacy and safety of the method. This encouraged us to conduct this follow-up study to fully evaluate the use of noncryopreserved fresh human fibroblast allografts in treating diabetic foot ulcers. Methods: Thirty-seven patients with diabetic foot ulcers were treated using fresh fibroblast allografts. Human dermal fibroblasts from healthy teenagers were cultured in DMEM/F-12 medium supplemented with 10% serum. The cultured cells were applied on the wounds immediately following debridement, with fibrin being used as a cell carrier. In eight weeks, percentages of complete healing, mean healing time, and patient satisfactions were assessed, with follow-up time ranging from 6 to 40 months. Results: Our study showed that 83.8% of the treated patients were complete healed. The time required for complete healing was $30.9{\pm}10.1$ days. Patient satisfaction scores for the experimental treatment were higher than those for the conventional method(mean scores of $8.1{\pm}1.1$ and $4.8{\pm}1.4$, respectively). No adverse events related to the study treatment occurred. Conclusion: The use of fresh human fibroblast allografts was found to be a safe and effective treatment for diabetic foot ulcers.

• Journal of Animal Reproduction and Biotechnology
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• v.37 no.4
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• pp.239-245
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• 2022

Protein and peptide candidates are screened to apply therapeutic application as a drug. Ensuring that these candidates are delivered and maximized effectiveness is still challenging and a variety of studies are ongoing. As drug delivery system vehicles, cell-penetrating peptide (CPP) can deliver various kinds of cargo into the cell cytosol. In a previous study, we developed Ara27 CPP, which are a zinc knuckle family protein of Arabidopsis, and confirmed internalization in human dermal fibroblasts and human dental pulp stem cells at low concentration with short time treatment condition without any toxicity. Ara27, an amphipathic CPP, could be modified and utilized in the biomedical field excluding the risk of toxicity. Therefore, we would like to confirm the non-toxic induced penetrating ability of Ara27 in various cell lines. The purpose of this study was to screen the cell internalization ability of Ara27 in various cell lines and to confirm Ara27 as a promising core CPP structure. First, Ara27 was screened to confirm non-toxicity concentration. Then, fluorescence-labeled Ara27 was treated on human normal cell lines, cancer cell lines and animal cell lines to identify the cellular internalization of Ara27. Ara27 was well intracellular localized in all cell lines and the intensity of fluorescence was remarkably increased in time pass manner. These results indicate that Ara27 has the potential as a core structure for applications in various drug delivery systems.

• KSBB Journal
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• v.20 no.1 s.90
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• pp.40-45
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• 2005

The production of matrix metalloproteinases (MMPs) by the UV irradiated skin fibroblast and the degradation of exracellular matrix (ECM) by these enzymes is known as one of the main reasons of photoaging. In this study, to investigate the relationship between aging and Spatholobi caulis extract, we examined the effects of antioxidant, in vitro MMP inhibition and expression of UVA-induced MMP-1 in human dermal fibroblasts. Spathoiobi caulis extract was found to show scavenging activities of radicals and reactive oxygen species (ROS) with the $IC_{50}$ values of $45.81{\mu}g/ml$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $3.11{\mu}g/ml$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. Spatholobi caulis extract inhibited the activities of MMP-1 in a does-dependent manner and the IC50 value calculated from semi-log plots was $31.96{\mu}g/ml$. Also, UVA induced MMP expression was reduced $74.66\%$ by treatment with Spatholobi caulis extract, and MMP-1 mRNA expression was reduced in a dose-dependent manner. Therefore Spatholobi caulis extract was able to significantly inhibit MMP expression in protein and mRNA level. All these results suggested that Spatholobi caulis extract may act as an anti-aging agent by antioxidation and reducing UVA-induced MMP-1 production.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.22 no.2
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• pp.123-132
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• 2000

$TGF-{\beta}_1$ is a potent chemotactic factor for inflammatory cells and fibroblasts. It also stimulates the celluar source and components of extracellular matrix and the production of proteinase inhibitors. Collectively, these biologic activities lead to the accumulation and stabilization of the nascent matrix, which is vital to infection control. The objective of this study is to investigate production of $TGF-{\beta}$ in vitro fibroblast culture in the presence of Staphylococcus enterotoxin B(SEB) and/or lipopolysaccharide(LPS) and to elucidate the role of $TGF-{\beta}_1$ which may be responsible for infection control. The fibroblasts were originated from gingiva and facial dermis in 26 year-old male patient. In the presence of LPS($0.01{\mu}g$, $0.1{\mu}g$, $1.0{\mu}g$), SEB($0.01{\mu}g$, $0.l{\mu}g$, $1.0{\mu}g$) respectively, $cells(5{\times}10^3ml)$ were cultivated in vitro. At 1, 3, and 5 days after incubation, cells were counted. Also, $cells(2.5{\times}10^5ml)$ were cultivated in EMEM with LPS(0.01, 0.1 and $1.0{\mu}g$), SEB(0.01, 0.1 and $1.0{\mu}g$) respectively and $LPS(0.1{\mu}g)$ and $SEB(0.1{\mu}g)$ in combination for 24, 48, and 72 hours respectively. Culture supernatants were harvested at 1, 2, and 3 days after incubation period and triplicate culture supernatants were pooled and $TGF-{\beta}_1$ was assayed in duplicate. The results were as follows. 1. In gingival fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell Proliferation occurred very significantly since 3 days after incubation, compared with the control and the production of $TGF-{\beta}_1$ occurred very significantly at 1 day after incubation, compared with the control. 2. In facial dermal fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation occurred very significantly at 1 day after incubation, compared with the control. In SEB exposure, the production of $TGF-{\beta}_1$ was decreased very significantly at 1 day after incubation, compared with the control. However, in LPS, SEB and LPS exposure, the production of $TGF-{\beta}_1$ was increased very significantly at 1 day after incubation, compared with the control. In conclusion, the concentration of bacterial toxins and the incubation period correlated with cell proliferation and production of $TGF-{\beta}_1$ very significantly. The gingival and facial dermal fibroblasts have different phenotype each other The orchestrated understanding of fibroblast proliferation and $TGF-{\beta}_1$ production play an important part in host defense against the bacterial Infection and may prevent tissue necrosis such as necrotizing fasciitis and life-threatening syndrome such as multiple organ failure.

• Journal of Life Science
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• v.33 no.4
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• pp.305-314
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• 2023

Ultraviolet B (UVB) irradiation causes skin diseases by inducing cellular oxidative stress, photoaging, and inflammation. This study aimed to investigate the protective effects of morin against UVB-induced oxidative stress in normal human dermal fibroblasts (NHDFs). Morin has been reported to be a potential therapeutic candidate for oxidative stress-mediated diseases, neurodegenerative diseases, and inflammation. Since morin has been identified as a potential antioxidant, we speculated that morin could alleviate UVB-induced apoptosis in NHDFs. Cell viability and intracellular reactive oxygen species (ROS) levels were measured using the MTT assay, H2DCFDA, and the DHE staining method, respectively. Lipid peroxidation and protein carbonyl formation were tested using ELISA kits. DNA fragmentation and comet assay were used to assess DNA damage. Apoptotic bodies were analyzed using Hoechst 33342 staining and TUNEL assay. The expression of apoptosis-related proteins was examined using Western blot analysis. Morin showed a cyto-protective effect by scavenging UVB-induced ROS, increasing the expression of antioxidant-related proteins and inhibiting UVB-induced oxidative alterations such as lipid peroxidation, protein carbonylation, and DNA damage. Morin protects against UVB-induced cell apoptosis by inhibiting Bcl-2-associated X protein, caspase-9, and caspase-3 expression, while increasing the expression of the anti-apoptotic protein Bcl-2. These effects of morin were conferred through decreased phosphorylation of p38 and c-Jun N-terminal kinase 1/2. The results demonstrated that morin may be developed as a preventive/therapeutic drug to be used to prevent UVB-induced skin damage.

• Archives of Plastic Surgery
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• v.32 no.4
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• pp.529-532
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• 2005

Expanding cells ex-vivo is very important in tissue-engineering. Culture medium is usually supplemented with fetal bovine serum(FBS) in most of the experiments. However, cells grown in bovine serum media may posses the possibilities of disseminating bovine diseases and/or stimulating the patient's immune reactions. To overcome these problems, autologous or homologous serum should be used instead of the FBS. The purpose of this study is to compare cell proliferation and collagen synthesis depending on the kind of sera mixed on media and to provide a guideline on applying established experimental data to clinical cases. Human dermal fibroblasts were obtained from four patients. Five thousand cells per well in 96-well plates were incubated DMEM/F-12 Nutrient with varying serum mixture; 10% autologous serum, 10% homologous serum, and 10% FBS. Five days after incubation fibroblast proliferation and collagen production were determined by MTT assay and CICP enzyme immunoassay. The mean cell number were; $3.95{\times}10^4/well$, $2.97{\times}10^4/well$ and $2.30{\times}10^4/well$, respectively. The average amounts of collagen synthesized were; 238.13 ng/ml, 204.88 ng/ml, and 163.88 ng/ml in each. These results show that the use of human serum mixture may contribute to, not only preventing disseminated infection of bovine diseases. but also increase cell proliferation and collagen synthesis without simulating the patient's immune reactions.