• Archives of Plastic Surgery
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• v.42 no.1
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• pp.11-19
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• 2015

Background Wound healing is an interaction of a complex signaling cascade of cellular events, including inflammation, proliferation, and maturation. $K^+$ channels modulate the mitogen-activated protein kinase (MAPK) signaling pathway. Here, we investigated whether $K^+$ channel-activated MAPK signaling directs collagen synthesis and angiogenesis in wound healing. Methods The human skin fibroblast HS27 cell line was used to examine cell viability and collagen synthesis after potassium chloride (KCl) treatment by Cell Counting Kit-8 (CCK-8) and western blotting. To investigate whether $K^+$ ion channels function upstream of MAPK signaling, thus affecting collagen synthesis and angiogenesis, we examined alteration of MAPK expression after treatment with KCl (channel inhibitor), NS1619 (channel activator), or kinase inhibitors. To research the effect of KCl on angiogenesis, angiogenesis-related proteins such as thrombospondin 1 (TSP1), anti-angiogenic factor, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), pro-angiogenic factor were assayed by western blot. Results The viability of HS27 cells was not affected by 25 mM KCl. Collagen synthesis increased dependent on time and concentration of KCl exposure. The phosphorylations of MAPK proteins such as extracellular-signal-regulated kinase (ERK) and p38 increased about 2.5-3 fold in the KCl treatment cells and were inhibited by treatment of NS1619. TSP1 expression increased by 100%, bFGF expression decreased by 40%, and there is no significant differences in the VEGF level by KCl treatment, TSP1 was inhibited by NS1619 or kinase inhibitors. Conclusions Our results suggest that KCl may function as a therapeutic agent for wound healing in the skin through MAPK signaling mediated by the $K^+$ ion channel.

• Reproductive and Developmental Biology
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• v.31 no.3
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• pp.153-159
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• 2007

In this study, using FIV-based lentivirus vector system, we tried to express hG-CSF in tetracycline-controllable manner. hG-CSF influences the proliferation, differentiation, and survival of cells in the neutrophil lineage. To enhance stability and translation of hG-CSF transcript, WPRE sequence was also introduced into FIV-Tet-On vector at downstream region of either the hG-CSF gene or the sequence encoding rtTA. Primary culture cells (CEF, chicken embryonic fibroblast; PFF, procine fetal fibroblast) infected with the recombinant FIV were cultured in the medium supplemented with or without doxycycline for 48 hours, and induction efficiency was measured by comparing the hG-CSF gene expression level using quantitative real-time PCR, Western blot and ELISA. Higher hG-CSF expression and tighter expression control were observed from the vector in which the WPRE sequence was placed at downstream of the hG-CSF (in CEF) or rtTA (in PEE) gene. This FIV-Tet-On vector system may be helpful in solving serious physiological disturbance problems which has continuously hampered successful production of transgenic animals and gene therapy.

• KSBB Journal
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• v.24 no.4
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• pp.397-402
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• 2009

The Portulaca oleracea (P. oleracea) is a popular herbal medicine in East Asia that was known to possess detoxification, antifebrile and antifungal effects. In the present study, we examined the biological activities of ethanol extracts of P. oleracea under various conditions with NIH3T3, B16F10, and MCF-7 cell line model systems. Extracts of P. oleracea (0.5 mg/ml) showed inhibition of expression of tyrosinase, but does not suppress either of TYRP-1 or DCT expression on B16F10 cells. Extracts of P. oleracea (2 mg/ml) showed anti-inflammatory effects on TNF-$\alpha$-stimulated NIH3T3/$NF{\kappa}B$-Luc cells and increase of the synthesis of collagen on NIH3T3 (wild type) cells. These results suggest that extracts of P. oleracea could be used as a functional biomaterial in developing a skin whitening agent and having the anti-inflammatory, anti-wrinkle, and anti-aging activities.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.27 no.2
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• pp.45-56
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• 2001

Korea mountain ginseng known as oriental miracle drug is an important medicinal plant. The effect of mountain ginseng adventitious roots extract has been described. The valuable root of mountain ginseng contained several kinds of ginsenosides that have been confirmed to have many active functions for the human body. However, the study of mountain ginseng has a limit because the price of wild ginseng is very expensive and rare. The mountain ginseng adventitious roots were derived from mountain ginseng callus that were induced from mountain ginseng roots. Adventitious roots were separated from callus and grown in solid media(Murachige and stoog media). It was cultured in a 20L bioreactor. After culturing for 40days, adventitious roots were harvested. Afterwards the harvested mountain ginseng adventitious roots were dryed and extracted. We examined the effect on melanogenesis of mountain ginseng adventitious roots extract. Here, we report the inhibitory effect of melanin biosynthesis on the adventitious roots extract of In vitro test. Also, we assessed the safety of adventitious roots extract. In vitro, cytotoxicity of adventitious roots extract was assessed in mouse fibroblast using two method: The neutral red uptake assay and the MTT assay. In vivo, the allergic and irritant were Patch teated in 30 patients. Consequently, extract of mountain ginseng adventitious roots have inhibitory effect on melanin biosynthesis in B-16 melanoma cell test, tyrosinase inhibitory test and DOPA auto-oxidation test. There were decreased 86%(0.5% concentration), 45%(1% concentration) and 61%(1% concentration), respectively.

• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.157-162
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• 2006

Endogenous 84 amino acid parathyroid hormone(PTH) is synthesized as a pre-pro hormone by the chief cells of the parathyroid glands. Physiological actions of PTH include regulation of bone metabolism, renal tubular reabsorption of calcium and phosphate, and intestinal calcium absorption. In addition, PTH stimulates new bone formation by extraordinary stimulation of osteoblastic activity and decreasing calcium excretion by the kidney. In this study, we constructed and tested retrovirus vectors designed to express the human parathyroid hormone(hPTH) gene under the control of the tetracycline-inducible promoters. To increase the hPTH gene expression at turn-on state, woodchuck hepatitis virus posttranscriptional regulatory element(WPRE) sequence was also introduced into retrovirus vector at downstream region of either the hPTH gene or the sequence encoding reverse tetracycline-controlled transactivator(rtTA). Transformed primary culture cells(porcine fetal fibroblast, PFF, chicken embryonic fibroblast, CEF) were cultured in the medium supplemented with or without doxycycline(tetracycline derivative) for 48 hours, and induction efficiency was measured by comparing the hPTH gene expression level using two step RT-PCR and ELISA Higher hPTH expression($3{\tims}10^4\;pg/ml,\;5.3{\times}10^4\;pg/ml$) and tighter expression control(up to 8 fold) were observed from the vector in which the WPRE sequence was placed at downstream of the hPTH gene. The resulting tetracycline inducible vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.36 no.1
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• pp.79-87
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• 2010

Ultraviolet is the one of the main environmental factors promoting aging process via increased intracellular generation of reactive oxygen species (ROS) and decreased expression of endogenous antioxidant enzymes and molecules. Therefore, in this study, we tried to search for natural skin-protective antioxidant materials from marine origins (Porphyra Thalli, Laminariae japonicae thallus, Ostreae Concha, Sargassum Thallus, Undaria thallus, Haliotidis Concha, Codium thalli, Syngnathoides biaculeatus, Hippocampus, Stichopus Stichopus, Thalli, Hizikia fusiforme thalli) which exhibit free radical scavenging activity and protect against UVB-induced cytotoxicity and oxidative cell death. Free radical scavenging activity was shown in order of Undaria thallus. Sargassum Thallus, Laminariae japonicae thallus, Hippocampus, Haliotidis Concha, Ostreae Concha, Syngnathoides biacuJeatus. In another experiment, UVB-induced cytotoxicity and cell death were effectively suppressed by treatment of Sargassum Thallus, Haliotidis Concha, Codium thalli, or Hippocampus water extract. Furthermore, UVB-induced cell death was mediated by intracellular accumulation or ROS, which was significantly inhibited by treatment with aforementioned extracts. The protective effect of these marine natural products seemed to be mediated by increased expression of antioxidant enzymes such as catalase, superoxide dismutase, and heme oxygenase-1. These results suggest that Sargassum Thallus, Haliotidis Concha, Codium thalli, and Hippocampus may have preventive and protective potentials as new functional cosmetics against oxidative stress-mediated skin damages and aging with antioxidant properties.

• Journal of Periodontal and Implant Science
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• v.49 no.1
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• pp.2-13
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• 2019

Purpose: The aim of this study was to conduct a histologic evaluation of irradiated calvarial defects in rats 4 weeks after applying fibroblast growth factor-2 (FGF-2) with hyaluronan or biphasic calcium phosphate (BCP) block in the presence or absence of adjunctive hyperbaric oxygen (HBO) therapy. Methods: Twenty rats were divided into HBO and non-HBO (NHBO) groups, each of which was divided into FGF-2 and BCP-block subgroups according to the grafted material. Localized radiation with a single 12-Gy dose was applied to the calvaria of rats to simulate radiotherapy. Four weeks after applying this radiation, 2 symmetrical circular defects with a diameter of 6 mm were created in the parietal bones of each animal. The right-side defect was filled with the materials mentioned above and the left-side defect was not filled (as a control). All defects were covered with a resorbable barrier membrane. During 4 weeks of healing, 1 hour of HBO therapy was applied to the rats in the HBO groups 5 times a week. The rats were then killed, and the calvarial specimens were harvested for radiographic and histologic analyses. Results: New bone formation was greatest in the FGF-2 subgroup, and improvement was not found in the BCP subgroup. HBO seemed to have a minimal effect on new bone formation. There was tendency for more angiogenesis in the HBO groups than the NHBO groups, but the group with HBO and FGF-2 did not show significantly better outcomes than the HBO-only group or the NHBO group with FGF-2. Conclusions: HBO exerted beneficial effects on angiogenesis in calvarial defects of irradiated rats over a 4-week healing period, but it appeared to have minimal effects on bone regeneration. FGF-2 seemed to enhance new bone formation and angiogenesis, but its efficacy appeared to be reduced when HBO was applied.

• Journal of Physiology & Pathology in Korean Medicine
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• v.36 no.4
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• pp.125-129
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• 2022

The purpose of study is to evaluate in vitro anti-oxidant and anti-inflammatory effects of Moringa Folium and Eucommiae Cortex 2:1 (g/g) mixtures (MEMix). HaCaT and human normal dermal fibroblast were treated with 0.01-1 mg/mL of MEMix to monitor cytotoxicity. Radical scavenging activities of MEMix were examined by DPPH assay. To explore anti-inflammatory effect, Raw 264.7 cells were pretreated with MEMix for 1h and subsequently exposed to LPS for 18h. NO release and cytotoxicity of Raw 264.7 cells were measured by adding Griess and MTT reagents, respectively. TNF-α, IL-1β, IL-6, and PGE2 productions were examined by ELISA. Immunoblot analysis was conducted to examine COX-2 expression in MEMix pretreated Raw 264.7 cells. Up to 1 mg/mL concentration, treatment of MEMix for 24 h did not affect normal dermal fibroblast viability and significantly reduced cell viability of HaCaT cells with no concentration dependency. MEMix increased DPPH radical scavenging activity with concentration dependency. Radical scavenging activities by 1 mg/mL of MEMix was comparable with 30 µM of trolox. Pretreatment of MEMix did not change the reduction of Raw 264.7 cell viability. Exposure of LPS in Raw 264.7 cells significantly increased NO, TNF-α, IL-1β, IL-6, and PGE2 productions, and MEMix pretreatment attenuated these productions by LPS concentration dependently. However, pretreatment with MEMix did not change COX-2 expression by LPS in Raw 264.7 cells. MEMix showed in vitro anti-oxidant and anti-inflammatory activities. MEMix would be useful candidate agent against inflammation.

• Microbiology and Biotechnology Letters
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• v.40 no.2
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• pp.117-127
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• 2012

The beneficial effects of adipose-derived stem cell conditioned media (ADSC-CM) for skin regeneration have previously been reported, despite the precise mechanism of how ADSC-CM promotes skin regeneration remaining unclear. ADSC-CM contains various secretomes and this may be a factor in it being a good resource for the treatment of skin conditions. It is also known that ADSC-CM produced in hypoxia conditions, in other words Advanced Adipose-Derived Stem cell Protein Extract (AAPE), has excellent skin regenerative properties. In this study, a human primary skin cell was devised to examine how AAPE affects human dermal fibroblast (HDF) and human keratinocyte (HK), which both play fundamental roles in skin regeneration. The promotion of collagen formation by HDFs was observed at 0.32 mg/ml of AAPE. AAPE treatment significantly stimulated stress fiber formation. DNA gene chips demonstrated that AAPE in HKs (p<0.05) affected the expression of 133 identifiable transcripts, which were associated with cell proliferation, migration, cell adhesion, and response to wounding. Twenty five identified proteins, including MMP, growth factor and cytokines such as CD54, FGF-2, GM-CSF, IL-4, IL-6, VEGF, TGF-${\beta}2$, TGF-${\beta}3$, MMP-1, MMP-10, and MMP-19, were contained in AAPE via antibody arrays. Thus, AAPE might activate the HK biological function and induce the collagen synthesis of HDF. These results demonstrate that AAPE has the potential to be used for clinic applications aimed at skin regeneration.

• The Journal of Advanced Prosthodontics
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• v.6 no.5
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• pp.406-414
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• 2014

PURPOSE. The objective of this study was to investigate the biologic effects of enamel matrix derivative (EMD) with different concentrations on cell viability and the genetic expression of human gingival fibroblasts (HGF) to zirconia surfaces. MATERIALS AND METHODS. Immortalized human gingival fibroblasts (HGF) were cultured (1) without EMD, (2) with EMD $25{\mu}g/mL$, and (3) with EMD $100{\mu}g/mL$ on zirconia discs. MTT assay was performed to evaluate the cell proliferation activity and SEM was carried out to examine the cellular morphology and attachment. The mRNA expression of collagen type I, osteopontin, fibronectin, and TGF-${\beta}1$ was evaluated with the real-time polymerase chain reaction (RT-PCR). RESULTS. From MTT assay, HGF showed more proliferation in EMD $25{\mu}g/mL$ group than control and EMD $100{\mu}g/mL$ group (P<.05). HGFs showed more flattened cellular morphology on the experimental groups than on the control group after 4h culture and more cellular attachments were observed on EMD $25{\mu}g/mL$ group and EMD $100{\mu}g/mL$ group after 24h culture. After 48h of culture, cellular attachment was similar in all groups. The mRNA expression of type I collagen increased in a concentration dependent manner. The genetic expression of osteopontin, fibronectin, and TGF-${\beta}1$ was increased at EMD $100{\mu}g/mL$. However, the mRNA expression of proteins associated with cellular attachment was decreased at EMD $25{\mu}g/mL$. CONCLUSION. Through this short term culture of HGF on zirconium discs, we conclude that EMD affects the proliferation, attachment, and cell morphology of HGF cells. Also, EMD stimulates production of extracellular matrix collagen, osteopontin, and TGF-${\beta}1$ in high concentration levels. CLINICAL RELEVANCE. With the use of EMD, protective barrier between attached gingiva and transmucosal zirconia abutment may be enhanced leading to final esthetic results with implants.