• 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.

• Korean journal of food and cookery science
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• v.24 no.6
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• pp.729-734
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• 2008

The principal objective of this study was to assess the possibility of transforming platycodin glycosides using various strains of probiotic bateria and edible fungi. Among the experimental microorganisms assess herein, Aspergillus niger KCTC 6909 evidenced the highest level of platycodin glycoside hydrolysis during fermentation. Particularly in cases in which the organism was incubated in the presence of rhamnose and platycodins. In order to produce the enzyme from Aspergillus niger effectively, various incubation conditions were assessd in order to determine the optimal conditions. The cytotoxicity on V79-4 (Chinese- hamster lung fibroblasts, normal cells) of platycodin was reduced significantly after conversion (concentration on $500{\mu}g/mL$, $1000{\mu}g/mL$); DPPH radical scavenging activity before conversion was 35.05%, and was 57.44% afterward. We noted significantly higher conversion activity inhibiting oxidative degradation. In conclusion, these results indicate that the proper combination of food microorganisms -and fermentation conditions can result in an increase in the glycoside hydrolysis of platycodin the resultant products of which reduce cytotoxicity- and increase anti-oxidant activity.

• Journal of the Korean Society of Laryngology, Phoniatrics and Logopedics
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• v.32 no.1
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• pp.15-23
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• 2021

Background and Objectives During speech, the vocal folds oscillate at frequencies ranging from 100-200 Hz with amplitudes of a few millimeters. Mechanical stimulation is an essential factor which affects metabolism of human vocal folds. The effect of mechanical vibration on the cellular response in the human vocal fold fibroblasts cells (hVFFs) was evaluated. Materials and Method We created a culture systemic device capable of generating vibratory stimulations at human phonation frequencies. To establish optimal cell culture condition, cellular proliferation and viability assay was examined. Quantitative real time polymerase chain reaction was used to assess extracellular matrix (ECM) related and growth factors expression on response to changes in vibratory frequency and amplitude. Western blot was used to investigate ECM and inflammation-related transcription factor activation and its related cellular signaling transduction pathway. Results The cell viability was stable with vibratory stimulation within 24 h. A statistically significant increase of ECM genes (collagen type I alpha 1 and collagen type I alpha 2) and growth factor [transforming growth factor β1 (TGF-β1) and fibroblast growth factor 1 (FGF-1)] observe under the experimental conditions. Vibratory stimulation induced transcriptional activation of NF-κB by phosphorylation of p65 subunit through cellular Mitogen-activated protein kinases activation by extracellular signal regulated kinase and p38 mitogen-activated protein kinases (MAPKs) phosphorylation on hVFFs. Conclusion This study confirmed enhancing synthesis of collagen, TGF-β1 and FGF was testified by vibratory stimulation on hVFFs. This mechanism is thought to be due to the activation of NF-κB and MAPKs. Taken together, these results demonstrate that vibratory bioreactor may be a suitable alternative to hVFFs for studying vocal folds cellular response to vibratory vocalization.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.11
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• pp.1492-1498
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• 2009

In the dermis, fibroblast plays an important role in the turnover of the dermal extracellular matrix. Collagen I and III, which are the most important dermal proteins of the extracellular matrix, function as a stabilizing scaffold of dermal connective tissues, as well as a regulator of differentiation and migration of dermal cells. In this study, we investigated the effect of various nutrients, such as ascorbic acid, silicon, Fe, lysine and proline which function as cofactors or building blocks on collagen synthesis. When the physiological concentrations of ascorbic acid (0-100 ${\mu}M$), silicon (0-50 ${\mu}M$), Fe (0-50 ${\mu}M$), lysine (0-150 ${\mu}M$) and proline (0-300 ${\mu}M$) were treated at HS27 for either 3 or 5 days, 5 day treatment of ascorbic acid at the low concentration (5-10 ${\mu}M$) increased the expression of collagen I and III protein by 115-1300% without increasing cell proliferation. 3 or 5 days treatment of Fe increased the expression of collagen I and III proteins up to 323% in parallel with cell proliferation by 164%. However, cell proliferation and expression of collagen I and III protein in silicon treated HS27 did not differ. Proline and lysine only increased cell proliferation up to 247.9%. Taken together, we demonstrate that the physiological concentrations of ascorbic acid and Fe enhance the expression of collagen I and III protein for treatment of 3 or 5 days.

• Applied Microscopy
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• v.30 no.2
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• pp.213-232
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• 2000

The development of the knee joint was studied by electron microscopy in human fetuses ranging from 20 mm to 260 mm crown-rump length ($7\sim30$ weeks of gestational age). The appearance of the primordium of the meniscus and cruciate ligament was conspicuous as the mesenchymal cells , preceeding that of joint space at 30 mm fetus. The primitive joint cavity was first seen in the interzone from the 40 mm fetus and its intermediate layer proceeded developing as a narrow cleft which was closely incorporated with two chondrogenic layers. Poorly differentiated mesenchymal cells of the meniscus at 40 mm fetus containing predominantly free ribosomes differentiated into fibroblasts at 60 mm fetus. By 100 mm fetus, the fibroblast in inner zone of the meniscus presented as oval profiles with a short cell processes, whereas middle and peripheral zones presented as elongated cells. Differentiation of the synovial membrane coincided with clarification of the joint cavity When dilatation of the synovial cavity occurred, the two types of synovial cells were identified at 60 mm fetus. By 100 mm fetus a majority of the intimal cells were B-type. B-type cells were clearly distinguishable from A-type cells by their content of extensive rough endoplasmic reticula and well developed Golgi complexes. In contrast, A-type cells had numerous filopodia, pinocytotic vesicles, lysosomes and large vacuoles. At 260 mm fetus the B-type cells were also a majority of intimal cells. At 260 mm fetus the inner zone of the meniscus was filled with parallel oriented fascicles of collagenous fibers and oval fibroblasts. The middle zone was constituted of parallel and radially arranged fibers and fibroblasts. The outer zone was populated by elongated fibroblasts encircled by crossed collagenous fibers with the blood vessels. At 30 mm fetus the fibroblasts of the cruciate ligament contained rough endoplasmic reticula and mitochondria. Collagen fibrils were noted within narrow cytoplasmic processes which were continued with the extracellular space. Collagen fibrils of ligament were filled in the bulk of extracellular space at 100 mm fetus. By $150\sim260mm$ fetus, the cruciate ligaments were constituted of longitudinally oriented bundle of collagen fibrils with irregular rows of round cells between.

• Development and Reproduction
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• v.12 no.1
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• pp.31-39
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• 2008

Neural tissue has limited intrinsic capacity of repair after injury, and the identification of alternate sources of neural stem cells has broad clinical potential. We isolated mesechymal-like stem cells from human adipose tissues (AT-MSCs), and studied on transdifferentiation-promoting conditions in neural cells. Dopaminergic and cholinergic neuron induction of AT-MSCs was also studied. Neural differentiation was induced by adding bFGF, EGF, dimethyl sulphoxide (DMSO) and butylated hydroxyanisole(BHA) in N2 Medium and N2 supplement. The immunoreactive cells for $\beta$-tubulin III, a neuron-specific marker, GFAP, an astrocyte marker, or Gal-C, an oligodendrocyte marker, were found. AT-MSCs treated with bFGF, SHH and FGF8 were differentiatied into dopaminergic neurons that were immunopositive for TH antibody. Differentiation of MSCs to cholinergic neurons was induced by combined treatment with basic fibroblast growth factor (bFGF), retinoic acid (RA) and sonic hedgehog (Shh). AT-MSCs treated with DMSO and BHA rapidly assumed the morphology of multipolar neurons. Both immunocytochemistry and RT-PCR analysis indicated that the expression of a number of neural markers including neuro D1, $\beta$-tubulin III, GFAP and nestinwas markedly elevated during this acute differentiation. While the stem cell markers such as SCF, C-kit, and Stat-3 were not expressed after preinduction medium culture, we confirmed the differentiation of dopaminergic and cholinergic neurons by TH/$\beta$-tubulin III or ChAT/ $\beta$-tubulin III positive cells. Conclusively, AT-MSCs can be differentiated into dopaminergic and cholinergic neuronsand these findings suggest that AT-MSCs are alternative cell source of treatment for neurodegenerative diseases.

• Journal of Periodontal and Implant Science
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• v.32 no.3
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• pp.445-456
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• 2002

The purpose of this study were to determine that dexamethasone(Dex) induces differentiation of periodontal ligament(PDL) cells to osteoblastic cells and to investigate expression of matrix Gla protein(MGP), which is one of bone matrix protein. The isolated human PDL cells and gingival fibroblasts were prepared and cultured. The fourth or sixth sub-passage cells were used in this experiments. control group, ascorbic acid and ${\beta}$-glycerophosphate treated group, ascorbic acid, ${\beta}$-glycerophosphate and l00nM Dex treated group, ascorbic acid, ${\beta}$-glycerophosphate, and 5 ${\mu}M$ Dex treated group were made for study. The results were as follows: Cellular morphological change of PDL cells according to time was investigated. At first, the cells exhibited confluent monolayer of spindle or polygonal appearance. The multilayer of cells were seen after 7 days of treatment. After 14 days, the cells lost polarity and were densely packed. The mineralized nodule formation was seen at 21 days in the only Dex treated PDL cell groups. In the gingival fibroblast groups and no Dex treated PDL cell groups, the mineralized nodule was not seen. The mineralized nodule formation of 5 ${\mu}M$ Dex treated group was higher than 100 nM Dex treated group. Alkaline phosphatase(ALP) activity was higher in the Dex treated PDL cell groups of 14 and 21 days than 0 and 7 days. MGP was expressed in the control and all experimental groups and the expression was constant at 0,7,14,21 day. The above results confirm that Dex is affected to differentiation of the PDL cells to osteoblastic or cementoblastic cells and has dose-dependent effect for mineralization. And, MGP is expressed in the PDL cells and is not affected to mineralization of PDL cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.32 no.5
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• pp.321-327
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• 2018

The purpose of this study was to investigate the effects of Cuscuta chinensis Lamark ethanol extract (CL) on wrinkle improvement. Cuscuta chinensis Lamark is known to contain dried saccharide, alkaloids, flavonoids, lignans and rein glycoside as major components of dried mature seeds of Cuscuta japonica Choisy. In this study, we evaluated the anti-wrinkle effects of CL and investigated bio-markers (e.g ; MMP-1, MMP-3, MMP-9, TIMP-1, type I procollagen) associated with skin wrinkle improvement. We tested the anti-wrinkle effect of CL using human fibroblast called CCD-986Sk cell. We observed an increase in MMPs, TIMP-1, and type 1 pro-collagen CL in CCD-986Sk cells irradiated with UVB at an intensity of $2mJ/cm^2$ for 60 seconds. As a result, CL decreased UVB-induced MMPs levels and mRNA expressions in CCD-986Sk cell. The levels and mRNA expressions of type I procollagen and TIMP-1 were increased by CL. These results suggest that CL has activities on improvement of skin wrinkle, which is induced by UVB radiation. Taken together, this study proposed the possibility of developing herbal medicine and functional herbal cosmetic materials with wrinkle-improving effects of Cuscuta chinensis Lamark.

• Journal of Periodontal and Implant Science
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• v.34 no.2
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• pp.317-332
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• 2004

Osteoblasts from alveolar bone may have an important role in the bone regeneration for periodontium, but their culture and characterization are not determined yet. The purpose of this study was to investigate the biological characteristics of primary explant cultured osteoblasts(PECO) from alveolar bone. Osteoblasts were isolated and cultured from alveolar socket of extracted tooth in children. To compare the characteristics, osteoblasts and gingival fibroblasts were cultured with DMEM at $37^{\circ}C$, 5% $CO_2$, l00% humidity incubator, and human fetal osteoblasts cell line(hFOB1) were cultured with DMEM at $34^{\circ}C$, 5%, $CO_2$ 100% humidity incubator. To characterize the isolated bone cells, morphologic change, cell proliferation and differentiation were measured. Morphology of PECO was small round body or cuboidal shape on inverted microscope and was similar with hFOB1. PECO became polygonal shape with stellate and had an amorphous shape at 9th passage in culture. PECO had significantly higher activity than that of gingival fibroblasts and hFOB1 in alkaline phosphatase activity. The expression of osteocalcin and bone sialoprotein in PECO was notably increased when compared with hFOB1 and gingival fibroblasts. These result indicated that PECO from alveolar bone in children has an obvious characteristics of osteoblast, maybe applied for the regeneration of bone.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.20 no.1
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• pp.1-13
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• 1994

An antioxidative compound was isolated from pine needles. This compound was identified as 4-hydroxy-5-methyl-3[2H]-furanone on the basis of spectroscopic evidences. It scavenged 1,1-diphenyl-2-picrylhydrazyl free radicals more efficiently than maltol and tocopherol did. It exhibited an inhibitory effect on the lipid peroxidation of rat liver microsome induced by Fe(ll)/ascorbate, and the protective effect against UV cytotoxicity in cultured human fibroblasts. In addition, HMF appeared to prevent the cellular melanogenesis in the cultured murine melanoma cells, more effectively than kojic acid, a well known inhibitor of melanogenesis, while the former was not so effective as the latter for the inhibilion of the tyrosinase. Considering that cellular melanogenesis is a metabolic process triggered by oxidative stress, it was tentatively deduced that the antioxidative property of HMF may afford the effect against cellular pigmentation by alleviating the causative stress. This study provided a novel inhibitor of melanogenesis, that might be useful for the cosmetic applications.