• Journal of Microbiology and Biotechnology
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• v.7 no.4
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• pp.267-271
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• 1997

We developed a dermal equivalent (DE) which was engineered using human dermal fibroblasts and a matrix of collagen gel. The in vitro construction of the DE was accomplished by casting a porcine collagen type I solution plus concentrated medium with isolated and cultured fibroblasts. These constructs were attached to culture dishes or left floating in culture medium. Contraction of attached gels results in decreased gel thickness without a change in gel diameter, and contraction of floating gels results in decreased gel thickness and diameter. After contraction, there was no increase in cell number in floating gels, but cells in attached gels began to increase after about 4 days of the lag phase in cell growth curve. At this lag phase, addition of fibroblast growth factor (FGF) at a concentration of $0.1{\mu}$/ml promoted cell proliferation in the attached collagen gels, but no effect in floating gels. These results indicate that the method of contraction had an influence on the extracellular matrix (ECM) organization, and this influenced not only cell growth but also fibroblast responsiveness to FGF. This suggests that attached collagen gel is more suitable as a dermal equivalent than the floating gel. And the final contracted area of attached gel is much larger than that of the floating gel since floating gel is contracted in all directions but attached gel is contracted only vertically.

• The Journal of the Korean Society for Microbiology
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• v.17 no.1
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• pp.7-14
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• 1982

A model of induction of neoplasia by viruses has develpoed from experimental studies in animals and in cultured cells and oncogenic transformation of cells is the result of integration of viral genetic information into the cellular DNA. The evidence for these associations was derived primarily from seroepidemiologic investigation. However, data indicating that the relation between HSV-2 and cervical cancer fits the model derived from experimental animal studies are not yet sufficient to draw conclusion with regard to the etiologic role the virus in the development of the neoplasms. In other hand, the K562 tumor cell is highly susceptible target for natural killer cell lysis by the lymphocytes of human and murine periperal blood. The characteristics of this effector cell type has been investigated. A study on natural killer cell mediated cytotoxicity(NKMC) against $^{51}Cr$-K562 as target cell was studed in HSV-2 infected ICR mouse. We have studied for susceptibility of HSV-2 against mouse embryo fibroblast(MEF) cells and NKMC from HSV-2 infected mouse. The results obtained that the mouse embryo fibroblast cells culture, the number and size of the cells were markedly increased and formed a monolayers relatively rapid, and become complete monolayer sheet around 72 hrs. Duration of cytopathic effect on MEF cells was rapid by serial passing of HSV-2. The morphology of the HSV-2 infected cells appear to be mainly round, ovium, spindle form and some of them was forming large giant cells. The NKMC was decrease in mouse with HSV-2 and comparison between effector/target cells ratio as 25:1 and 50:1 respectively, the NKMC was found to be more significantly decreased than normal control we have concluded that the natural killer cell activity of the viral infected mouse was shown as a suppressed during the HSV-2 infection, day 7th and 14th.

• The Korean Journal of Physiology and Pharmacology
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• v.21 no.3
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• pp.327-334
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• 2017

Epidemiologic interest in particulate matter (PM) is growing particularly because of its impact of respiratory health. It has been elucidated that PM evoked inflammatory signal in pulmonary epithelia. However, it has not been established $Ca^{2+}$ signaling mechanisms involved in acute PM-derived signaling in pulmonary fibroblasts. In the present study, we explored dust particles PM modulated intracellular $Ca^{2+}$ signaling and sought to provide a therapeutic strategy by antagonizing PM-induced intracellular $Ca^{2+}$ signaling in human lung fibroblasts MRC5 cells. We demonstrated that PM10, less than $10{\mu}m$, induced intracellular $Ca^{2+}$ signaling, which was mediated by extracellular $Ca^{2+}$. The PM10-mediated intracellular $Ca^{2+}$ signaling was attenuated by antioxidants, phospholipase blockers, polyADPR polymerase 1 inhibitor, and transient receptor potential melastatin 2 (TRPM2) inhibitors. In addition, PM-mediated increases in reactive oxygen species were attenuated by TRPM2 blockers, clotrimazole (CLZ) and N-(p-amylcinnamoyl) anthranilic acid (ACA). Our results showed that PM10 enhanced reactive oxygen species signal by measuring DCF fluorescence and the DCF signal attenuated by both TRPM2 blockers CLZ and ACA. Here, we suggest functional inhibition of TRPM2 channels as a potential therapeutic strategy for modulation of dust particle-mediated signaling and oxidative stress accompanying lung diseases.

• Proceedings of the SCSK Conference
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• 2003.09a
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• pp.35-49
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• 2003

In this study, in an attempt to search for functional cosmetic ingredients from higher fungal, we have produced exopolysaccharides (GF-l, approximately carbohydrate 75％, protein 25％) and polysaccharide (GF-2) of mycelium extract, by submerged culture of Grifolafrondosa. For applications in anti-aging cosmetic field, we investigated the diverse biological activities. Antioxidant activity and inhibition of Matrixmetalloproteinases (MMPs) were investigated enzymatic assays by measuring the superoxide scavenging activity using xanthine-xanthine oxidase system and the proteolytic activity of MMPs using EnzChek Collagenase/Gelatinase kits, respectively. GF-l polysacchairde showed inhibition of superoxide radical by 90％ at a concentration of 0.2％ (w/v) and inhibition of collagenase by 45％ at 0.2％ (w/v). GF-2 polysaccharide of mycelium extract also exhibited good antioxidant activity. However, MMPs inhibition activity was relatively lower level compared to GF-l polysaccharides. The treatment of human dermal fibroblast (HDF) with GF-l and GF-2 polysaccharides increased the proliferation of fibroblast by approximately 23-25％ at a concentration of 0.5％ (w/v), also showed collagen synthesis increase in HDF by about 50％ at 0.5％ (w/v) compared to that of untreated control. We also report the moisturizing effects of polysaccharides in cosmetic products (O/W emulation) and its own ingredient, in vitro and in vivo. The GF-1 polysaccharide showed higher moisturizing ability than sodium hyaluronate, which is the most commonly used moisturizers ingredient. These results suggest the GF-l polysaccharide, protein-bound polysaccharide, may be used as an ingredient for new moisturizing and anti-aging cosmeceuticals.

• The Journal of Advanced Prosthodontics
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• v.5 no.3
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• pp.341-350
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• 2013

PURPOSE. To evaluate adherence of human gingival fibroblasts (HGFs) to transmucosal abutment of dental implant with different surface conditions with time and to investigate the roles of focal adhesion linker proteins (FALPs) involved in HGFs adhesion to abutment surfaces. MATERIALS AND METHODS. Morphologies of cultured HGFs on titanium and ceramic discs with different surface were observed by scanning electron microscopy. Biocompatibility and focal adhesion were evaluated by ultrasonic wave application and cell viability assay. FALPs expression levels were assessed by RT-PCR and western blot. RESULTS. There seemed to be little difference in biocompatibility and adhesion strength of HGFs depending on the surface conditions and materials. In all experimental groups, the number of cells remaining on the disc surface after ultrasonic wave application increased more than 2 times at 3 days after seeding compared to 1-day cultured cells and this continued until 7 days of culture. FALPs expression levels, especially of vinculin and paxillin, also increased in 5-day cultured cells compared to 1-day cultured fibroblasts on the disc surface. CONCLUSION. These results might suggest that the strength of adhesion of fibroblasts to transmucosal abutment surfaces increases with time and it seemed to be related to expressions of FALPs.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.429-432
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• 2000

Chitosan scaffold is widely applied to drug delivery and tissue engineering. We have developed chitosan scaffolds, with various pore size, by differing freezing temperature and duration of ultraviolet (UV) irradiation, for reconstructing skin equivalent. Chitosan scaffold was coated with type I collagen, fibronectin and basic fibroblast growth factor (bFGF) in various combinations and concentrations, to evaluate the effect of extracellular matrix (ECM) and bFGF on cell adhesion, growth and differentiation of dermal fibroblasts. Human dermal fibroblasts, isolated from newborn foreskin and passaged between 3 and 5, were seeded on the top of scaffolds and cultivated for 2 weeks. We examined the morphology and the secretion of ECM of fibroblasts using scanning electron microsopy (SEM) and histochemistry. A stellate morphology of fibroblasts were seen in all groups. The scaffold coated with either type I collagen and bFGF or type I collagen and fibronectin, however, showed the best condtion of dermal fibroblasts, in that the highest cell number and ECM secretion were seen. On the contrary, scaffolds coated with all three factors, type I collagen, bFGF and fibronectin, showed lower number of cells and ECM secretion than scaffolds with two factors. There was a tendency of dose-dependence in all three factors for fibroblast growth and ECM secretion. In conclusion, we may suggest that chitosan scaffold coated with either type I collagen/bFGF or type I collagen/fibronectin could provide more favorable environment for the growth and differentiation of dermal fibroblasts.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.11
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• pp.1551-1558
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• 2008

The mammalian cellular retinoic acid-binding proteins, CRABP-I and CRABP-II, bind retinoic acid which acts as an inducer of differentiation in several biological systems. To investigate a possible role for CRABP-II in bovine adipogenesis, we have cloned bovine CRABP-II cDNA and the coding region for CRABP-I. The predicted amino acid sequences of CRABP-II were highly conserved among several animal species (human, mouse, and rat at 97%, 93%, and 93%, respectively). The expression pattern of bovine CRABP-II was examined in greater details by applying RT-PCR to various bovine tissues. CRABP-II mRNA was expressed in most adipose-containing tissues. Moreover, the expression of CRABP-I and -II mRNA dramatically increased during the differentiation of adipocytes from bovine intramuscular fibroblast-like cells. The effects of retinoic acid on adipocyte differentiation of bovine intramuscular fibroblast-like cells were concentration-dependent. Retinoic acid activated the formation of lipid droplets at a level of 1 nM, whereas inhibition was observed at a level of $1{\mu}M$. CRABP-I gene was up-regulated and CRABP-II gene down-regulated by retinoic acid during adipocyte differentiation. These results suggest that CRABPs may play an important role in the regulation of intracellular retinoic acid concentrations during adipogenesis.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.14 no.4
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• pp.1857-1862
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• 2013

Dermal fibroblasts produce the many components of the extracellular matrix (ECM) that are needed to maintain connective tissue integrity and repair tissue injuries. This study investigated the effects of Areca catechu extract (ACE) on dermal fibroblast cell activation. Cultured human dermal fibroblasts were treated with ACE, and then ECM production was determined by ELISA, Western blot and RT-PCR. ACE significantly accelerated the production of type 1 collagen, fibronectin, and transforming growth factor (TGF)-${\beta}1$ by ELISA and type 1 collagen by Western blot assay. ACE also increased the gene expression of COL1A1, TGF-${\beta}1$, keratinocyte growth factor (KGF) and insulin growth factor (IGF)-1. These results suggest that ACE has the potential to stimulate ECM production and that it might be suitable for maintaining skin texture.

• Journal of Ginseng Research
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• v.45 no.4
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• pp.510-518
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• 2021

Background: Gintonin is a newly derived glycolipoprotein from the roots of ginseng. The purpose of this study is to investigate the anti-arthritic efficacy of Gintonin on various proteases and inflammatory mediators that have an important role in arthritis. Methods: Fibroblast-like synoviocytes (FLS) were treated with Gintonin and stimulated with interleukin (IL)-1β 1 hour later. The antioxidant effect of Gintonin was measured using MitoSOX and H₂DCFDA experiments. The anti-arthritic efficacy of Gintonin was examined by analyzing the expression levels of inflammatory mediators using RT-PCR, western blot, and ELISA. The phosphorylation of mitogen-activated protein kinase (MAPK) pathways and translocation of nuclear factor kappa B (NF-κB)/p65 into the nucleus were also analyzed using western blot, ELISA, and immunocytochemistry. Collagen-induced arthritis (CIA) mice model was used. Mice were orally administered with Gintonin (25, 50, and 100 mg/kg) every 2 days for 45 days. The body weight, arthritis score, squeaking score, and paw volume were measured as the behavioral parameters. After sacrifice, H&E and safranin-O staining were performed for histological analysis. Results: Gintonin significantly inhibited the expression of inflammatory intermediates. Gintonin prevented NF-κB/p65 from moving into the nucleus through the JNK and ERK MAPK phosphorylation in FLS cells. Moreover, Gintonin suppressed the symptoms of arthritis in the CIA mice model. Conclusion: As a result, the antioxidant and anti-inflammatory effects of Gintonin were demonstrated, and ultimately the anti-arthritic effect was proved. Collectively, Gintonin has a great potential as a therapeutic agent for arthritis treatment.

• Journal of the Korea Institute of Information and Communication Engineering
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• v.25 no.2
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• pp.221-227
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• 2021

Recently, interests in mask-type skin care devices using light-emitting diodes have been increasing and optical stimuli at certain wavelengths have been known to have various therapeutic effects, such as skin whitening, acne treatment, elasticity and wrinkle improvement by controlling the exposure to wavelengths of light and irradiation time. In particular, light sources of different wavelengths are applied in masks for the purpose of suppressing skin aging, inducing cell proliferation, and alleviating skin inflammation. In this paper, we developed a light-emitting diode control system that is actively used in skin regeneration masks using a microcontroller. Optical stimuli with different manners were applied to skin fibroblast cells in a single or complex wavelengths, and then confirmed how they are effective in the cell differentiation. In addition, we found a specific wavelength that has a positive effect on cell proliferation rates, and confirm the effectiveness of cell proliferation by image processing based quantitative analysis.