Ultraviolet B (UVB) exposure is a risk factor for skin damage resulting in oxidative stress, inflammation, and cell death. The purpose of this study was to investigate the physicochemical properties of Platycodon grandiflorum (PG) to improve its biological activities using a three-step steaming process. We investigated the protective effects of PG and steamed PG extracts on human dermal fibroblasts (HDFs) against UVB radiation-induced oxidative stress and inflammation as well as the underlying mechanisms. The antioxidant potential of the PG extracts was evaluated by measuring the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) scavenging activity. ABTS and DPPH were shown by the 0, 30, and 70% ethanol extracts of 2S-PG and 3S-PG ($IC_{50}$, 28~45 and $27{\sim}30{\mu}g/mL$, respectively). Treatment of UVB-irradiated cells with steamed PG ($25{\sim}400{\mu}g/mL$) did not affect their viability. The streamed PG extract suppressed UVB-induced generation of reactive oxygen species (ROS). In addition, streamed PG extract reduced cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) protein expression in UVB-irradiated HDF, regulating nuclear factor $(NF)-{\kappa}B$ expression. These findings suggest that steamed PG extract may be potentially effective against inflammation associated with UVB-induced oxidation stress.
Journal of the Society of Cosmetic Scientists of Korea
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v.39
no.3
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pp.177-186
/
2013
Skin dermal fibroblast is the major collagen-producing cell type in human skin. As aging process continues in human skin, collagen production is reduced and fragmentation is increased, which is initiated by matrix metalloproteinase-1 (MMP-1). This imbalance of collagen homeostasis impairs the structure and function of dermal collagenous extracellular matrix (ECM), thereby promoting skin aging. Cysteine-rich protein 61 (CCN1), a member of the CCN family, negatively regulates collagen homeostasis in primary human skin dermal fibroblast cells. It is known in aging fibroblast cells that elevated CCN1 expression substantially reduces type I procollagen and concurrently increases MMP-1, which initiates fibrillar collagen degradation. And proliferation rate of aging fibroblast cells is reduced compared to the pre-aging fibroblast cells. In this study, we confirmed that the replicative senescence dermal fibroblast cells increased the expression levels of MMP-1 and decreased the production of type I procollagen. Our results also showed that the replicative senescence dermal fibroblast cells increased in the expression of CCN1 and decreased in the proliferation rate. Hydrolyzed Ulva pertusa extracts are the materials to improve photo-aging by reducing the expression of MMP-1 that was increased by ultraviolet and by promoting the synthesis of new collagen from fibroblast cells. In this study, we also investigated the hydrolyzed U. pertusa extract to see whether it inhibits CCN1 protein expression in the senescence fibroblasts. Results showed that the hydrolyzed U. pertusa extract inhibited the expression of MMP-1 and increased the production of type I procollagen in the aging skin fibroblast cells cultured. In addition, the proteins that regulate collagen homeostasis CCN1 expression were greatly reduced. The hydrolyzed U. pertusa extract increased the proliferation rate of the aging fibroblast cells. These results suggest that replicative senescent fibroblast cells may be used in the study of cosmetic ingredients as a model of the natural aging. In conclusion, the hydrolyzed U. pertusa extract can be used in anti-wrinkle functional cosmetic material to improve the natural aging skin care as well as photo-aging.
Mechanical stimulation is known to activate several cellular signal transduction pathways, leading to the induction of signaling molecules and extracellular matrix (ECM) proteins, thereby modulating cellular activities, such as proliferation and survival. In this study, primary human dermal fibroblasts (HDFs) were seeded onto chitosan-based scaffolds, and then cultured for 3 weeks in a bioreactor under a cyclic strain of 1 Hz frequency. Compared to control samples cultured under static conditions, the application of a cyclic strain stimulated the proliferation of HDFs in I week, and by week 3 the thickness of the cell/scaffold composites increased 1.56 fold. Moreover, immunohistochemical staining of the culture media obtained from the cell/scaffold samples subjected to the cyclic strain, revealed increases in the expression and secretion of ECM proteins, such as fibronectin and collagen. These results suggest that the preconditioning of cell/scaffold composites with a cyclic strain may enhance the proliferation of HDFs, and even facilitate integration of the engineered artificial dermal tissue into the host graft site.
Journal of the Society of Cosmetic Scientists of Korea
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v.41
no.1
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pp.1-7
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2015
Cornus walteri Wanger has been used in folk medicine in Korea. Ultraviolet (UV) irradiation has been known as a major cause of photo damage in skin. In the present study, research on how to cure damaged cells by UVB was conducted using an extract of Cornus walteri Wanger leaves (CWE), which was treated with an enzyme. CWE was applied to human dermal fibroblasts (HDFs) affected by UVB. UVB-irradiated HS68 cells showed increased caspase-3 activity, phosphorylation of p53, ${\gamma}H2AX$, cyclobutane pyrimidine dimers (CPDs) formation, and DNA fragmentation compared with non-irradiated cells. However, all these effects were inhibited by treatment with CWE for 12 h after UVB irradiation. Furthermore, CWE has proved not to cause primary skin irritation through the human patch test. Collectively, these results suggest that CWE could be a new potential candidate as photoprotective agent against UVB-induced cellular damage in HDFs.
Jang, Ik-Soon;Jo, Eunbi;Park, Soo Jung;Baek, Su Jeong;Hwang, In-Hu;Kang, Hyun Mi;Lee, Je-Ho;Kwon, Joseph;Son, Junik;Kwon, Ho Jeong;Choi, Jong-Soon
Journal of Ginseng Research
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v.44
no.1
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pp.50-57
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2020
Background: The cellular senescence of primary cultured cells is an irreversible process characterized by growth arrest. Restoration of senescence by ginsenosides has not been explored so far. Rg3(S) treatment markedly decreased senescence-associated β-galactosidase activity and intracellular reactive oxygen species levels in senescent human dermal fibroblasts (HDFs). However, the underlying mechanism of this effect of Rg3(S) on the senescent HDFs remains unknown. Methods: We performed a label-free quantitative proteomics to identify the altered proteins in Rg3(S)-treated senescent HDFs. Upregulated proteins induced by Rg3(S) were validated by real-time polymerase chain reaction and immunoblot analyses. Results: Finally, 157 human proteins were identified, and variable peroxiredoxin (PRDX) isotypes were highly implicated by network analyses. Among them, the mitochondrial PRDX3 was transcriptionally and translationally increased in response to Rg3(S) treatment in senescent HDFs in a time-dependent manner. Conclusion: Our proteomic approach provides insights into the partial reversing effect of Rg3 on senescent HDFs through induction of antioxidant enzymes, particularly PRDX3.
Cytokines are hormone-like proteins which mediate and regulate inflammatory and immune responses. Interleukin-6 (IL-6) is involved in the final differentiation of B cells into antibody-producing cells. Interleukin-8 (IL-8) is a neutrophil chemotactic factor that plays an important role in the recruitment of neutrophil to inflammatory loci. Inflammatory mediators by cells in the gingiva have been implicated in the initiation and progression of periodontitis and oral infection. The purpose of this study was conducted to investigate the effect of lipopolysaccharide (LPS), staphylococcus enterotoxin B (SEB) on production of IL-6 and IL-8 by human gingival and facial dermal fibroblasts. Primary cultured human gingival and facial dermal fibroblasts were incubated with LPS (0.01, 0.1, $1.0{\mu}g/ml$), SEB (0.01, 0.1, $1.0{\mu}g/ml$) or LPS $(0.1{\mu}g/ml)$ plus SEB $(0.1{\mu}g/ml)$. Culture supernatants were collected at 24, 48, and 72 hrs and assessed for IL-6 and IL-8 production by enzyme-linked immunosorbent assay. IL-6 production in gingival fibroblasts stimulated with LPS was higher than that with SEB. IL-6 production by double exposure with LPS plus SEB was amplified in comparison with single exposure of LPS or SEB. IL-6 production in facial dermal fibroblasts was increased only by stimulation with a high concentration of LPS $(1.0{\mu}g/ml)$. Its production in facial dermal fibroblasts by exposure with SEB was decreased in comparison with control, nontreated cells. Therefore, gingival fibroblasts showed higher sensitivity than facial dermal fibroblasts in response to low concentration of LPS. Also, IL-6 production by double exposure with LPS plus SEB was amplified in comparison with single exposure of LPS or SEB. IL-8 production in gingival fibroblasts was enhanced greatly only by stimulation of high concentration of LPS $(1.0{\mu}g/ml)$. That by exposure with SEB was increased only in 24 hrs cultivation. IL-8 production by double exposure with LPS plus SEB was amplified in comparison with single exposure of LPS or SEB. IL-8 production in facial dermal fibroblasts was decreased by LPS and increased only in 48 hrs cultivation by SEB. IL-8 production by double exposure with LPS plus SEB was enhanced only in 48 hrs cultivation in comparison with single exposure of LPS or SEB. therefore, IL-6 and IL-8 production were released at various quantities according to bacterial toxin applied and site of fibroblast harvested. These results suggest that gingival fibroblasts may be concerned with IL-6 and IL-8 related inflammatory response more than facial dermal fibroblasts.
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.
A novel retinol derivative, polyethoxylated retinamide (Medimin A) was synthesized, as an anti-aging agent. Collagen synthesis, skin permeation, stability, and toxicity of Medimin A were evaluated and compared with those of retinol and retinyl palmitate. In vitro collagen synthesis was evaluated by quantitative assay of [$^3H$]-proline incorporation into collagenase sensitive protein in fibroblast cultures. For in vitro skin permeation experiments, Franz diffusion cells (effective diffusion area: $1, 766{\;}\textrm{cm}^2$) and the excised skin of female hairless mouse aged 8 weeks were used The stabilities of retlnoids were evaluated at two different temperature ($25{\;}^{\circ}C$ and $40{\;}^{\circ}C$) and under UV in solubilized state and in OW emulsion. To estimate the safety, acute oral toxicity, acute dermal toxicity, primary skin irritation, acute eye irritation and human patch test were performed The effect of Medimin A on collagen synthesis was similar to that of retinol. The skin permeability of Medimin A was higher than those of retinol and retinyl palmitate. The Medimin A was more stable than retinol and retinyl palmitate. Medimin A was nontoxic in various toxicological tests. These results suggest that Medimin A would be a good anti-aging agent for enhancing bioavailability and stability.
Purpose : In this study, we intended to observe the anti-wrinkle and moisturizing effects of dried pomegranate juice concentration powder (PCP) using in vitro test. Materials and methods : Antioxidant effects of PCP were determined by free radical scavenging capacity (DPPH assay) and the cytotoxicity of PCP was examined in human keratinocyte (HaCaT) and human primary dermal fibroblast-neonatal (HDF) cells. To investigate the moisturizing effect of PCP, hyaluronan synthesis was examined in HaCaT cells. Activity of procollagen production were assessed in HDF cells and elastase inhibition properties of PCP were evaluated in cell free condition, to determine their anti-wrinkle effects. Metalloproteinase 1 (MMP-1) activity was also assessed following UVB irradiation, in the current in vitro experiment. Results : No PCP treatment related significant cytotoxic effects were demonstrated against to the both HDF and HaCaT cells. PCP showed favorable free radical scavenging activities in dose-dependent manner. In PCP-treated HaCaT cells, hyaluronan synthesis was non-significantly but markedly increased, and pro-collagen productions were significantly increased in HDF cells, at all three different concentrations (0.25, 0.75 and 1 mg/ml), and elastase inhibitory activities were observed by PCP treatment. A significant decrease in UVB-induced MMP-1 activity was also observed in 1 mg/ml PCP-treated HDF cells as compared to those of UVB-exposed cells. Conclusions : Taken together, these results suggest that PCP has favorable antioxidant, anti-wrinkle and moisturizing effects without meaningful cytotoxicity on HDF and HaCaT cell lines.
Journal of the Korea Academia-Industrial cooperation Society
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v.19
no.3
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pp.229-242
/
2018
The effects of orally administered fermented porcine placenta (FPP) and its major dipeptides, L-Leucyl-Glycine (Leu-Gly) and Glycyl-L-Leucine (Gly-Leu), on UVB-induced wrinkle formation of the skin in hairless mice was studied. Treatment with FPP, Leu-Gly or Gly-Leu increased type I procollagen synthesis and decreased MMP-1 (matrix metalloproteinase-1) in human dermal fibroblast cells (HDF-N). Hairless mice were also exposed UVB irradiation three times a week and fermented porcine placenta extract (FPP), Leu-Gly and Gly-Leu was administered once a day for eight weeks. Daily intake of FPP, Leu-Gly and Gly-Leu for eight weeks decreased wrinkles, erythema and thickness of the skin and increased skin hydration and synthesis of collagen relative to a UVB-control. Moreover, FPP, Leu-Gly or Gly-Leu intake decreased the expression of MMP-3 and MMP-13 mRNA levels and inhibited activation of MMP-2 and MMP-9 induced by UVB irradiation in hairless mice skin. These results suggest that major dipeptides of the placenta, Leu-Gly and Gly-Leu have the potential for use as a functional food ingredient with anti-wrinkling properties.
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