• Title/Summary/Keyword: Human Fibroblast

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The effects of chitosan on the human periodontal ligament fibroblasts in vitro (키토산이 치주인대 섬유아세포에 미치는 영향)

  • Paik, Jeong-Won;Lee, Hyun-jung;Yoo, Yun-Jung;Cho, Kyoo-Sung;Kim, Chong-Kwan;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.31 no.4
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    • pp.823-832
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    • 2001
  • Periodontal therapy has dealt primarily with attempts at arresting progression of disease, however, more recent techniques have focused on regenerating the periodontal ligament having the capacity to regenerate the periodontium. The effect of chitosan(poly-N-acetyl glucosaminoglycan), a carbohydrate biopolymer extracted from chitin, on periodontal ligament regeneration is of particular interest. The purpose of this study was to evaluate the effect of chitosan on the human periodontal ligament fibroblasts(hPDLFs) in vitro, with special focus on their proliferative properties by M'IT assay, the synthesis of type I collagen by reverse transcription-polymerase chain reaction(RT-PCR) and the activity of alkaline phosphatase(ALP). Fibroblast populations were obtained from individuals with a healthy periodontium and cultured with ${\alpha}MEM$ as the control group. The experimental groups were cultured with chitosan in concentration of 0.01,0.1, 1,2mg/ml. The results are as follows; 1. Chitosan-induced proliferative responses of hPDLFs reached a plateau at the concentration of O.lmg/ml(p<0.05). 2. When hPDLFs were stimulated with 0.lmg/ml chitosan, mRNA expression of type I collagen was up-regulated. 3. When hPDLFs were stimulated with 0.lmg/ml chitosan, ALP activity was significantly up-regulated(p<0.05). In summary, chitosan(0.lmg/ml) enhanced the type I collagen synthesis in the early stage, and afterwards, facilitated differentiation into osteogenic cells. The results of this in vitro experiment suggest that chitosan potentiates the differentiation of osteoprogenitor cells and may facilitate the formation of bone.

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Improvement of the Biocompatibility of Chitosan Dermal Scaffold by Rigorous Dry Heat Treatment

  • Kim, Chun-Ho;Park, Hyun-Sook;Gin, Yong-Jae;Son, Young-Sook;Lim, Sae-Hwan;Park, Young-Ju;Park, Ki-Sook;Park, Chan-Woong
    • Macromolecular Research
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    • v.12 no.4
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    • pp.367-373
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    • 2004
  • We have developed a rigorous heat treatment method to improve the biocompatibility of chitosan as a tissue-engineered scaffold. The chitosan scaffold was prepared by the controlled freezing and lyophilizing method using dilute acetic acid and then it was heat-treated at 110$^{\circ}C$ in vacuo for 1-3 days. To explore changes in the physicochemical properties of the heat-treated scaffold, we analyzed the degree of deacetylation by colloid titration with poly(vinyl potassium sulfate) and the structural changes were analyzed by scanning electron microscopy, Fourier transform infrared (FT-IR) spectroscopy, wide-angle X-ray diffractometry (WAXD), and lysozyme susceptibility. The degree of deacetylation of chitosan scaffolds decreased significantly from 85 to 30% as the heat treatment time increased. FT-IR spectroscopic and WAXD data indicated the formation of amide bonds between the amino groups of chitosan and acetic acids carbonyl group, and of interchain hydrogen bonding between the carbonyl groups in the C-6 residues of chitosan and the N-acetyl groups. Our rigorous heat treatment method causes the scaffold to become more susceptible to lysozyme treatment. We performed further examinations of the changes in the biocompatibility of the chitosan scaffold after rigorous heat treatment by measuring the initial cell binding capacity and cell growth rate. Human dermal fibroblasts (HDFs) adhere and spread more effectively to the heat-treated chitosan than to the untreated sample. When the cell growth of the HDFs on the film or the scaffold was analyzed by an MTT assay, we found that rigorous heat treatment stimulated cell growth by 1.5∼1.95-fold relative to that of the untreated chitosan. We conclude that the rigorous dry heat treatment process increases the biocompatibility of the chitosan scaffold by decreasing the degree of deacetylation and by increasing cell attachment and growth.

Effects of Sophorae Radix (SFR) on Allergic Inflammation related Atopy Dermatitis (고삼(苦蔘)이 알러지성 염증 반응에 미치는 영향)

  • Yu, Mao-Lun;Koo, Young-Sun;Kim, Dong-Hee
    • Journal of Haehwa Medicine
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    • v.15 no.2
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    • pp.135-148
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    • 2006
  • Sophorae Radix (SFR) is known as a therapeutic drug that has been used in Oriental traditional medicine for the treatment of skin and mucosal ulcers, gastrointestinal hemorrhage, diarrhea, inflammation and arrhythmia. In the present study, we examined the effects of the aqueous extract of SFR on anti-inflammation, anti-allergic and anti-oxidant effect in various cell lines; they include mouse lung fibroblast cells (hFCs), human mast cells (HMC-1), human monocytic cells (THP-1), and RAW 264.7 cells. Treatment with SFR extract at a concentration of 250 ${\mu}g$/ml for 24h showed no significant decrease in the survival rate of the hFCs. SFR decreased the mRNA expression of IL-8, TNF-$\alpha$, and IL-6 in HMC-1 cells. SFR extract treatment significantly inhi-bited the protein expression of IL-6 and, IL-8 induced by mite in THP-1 cells and it also did MCP-1 expression. We examined the alternation of histamine release in HMC-1 cells for investigating anti-allergic effect of SFR. Histamine secretion decreased after the treatment with SFR. In addition, SFR extract treatment at a concentration of 10 ${\mu}g$/ml, 100 ${\mu}g$ /ml, and 200 ${\mu}g$/ml lowered the $\beta$-hexosaminidase to 10.3%, 21.7%, and 50.8%, respectively. IC50 of SFR extract in RBL-2H3 cells was 196.85 ${\mu}g$/ml. Both activity of NF-$\kappa$B promoter in RBL-2H3 cells significantly diminished after the dose-dependent treatment of SFR. Therefore, our results indicate that SFR has anti-inflammatory and it may be useful for treating allergic diseases such as atopic dermatitis.

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Effects of Anthriscus sylvestris Hoffmann Extract on the Biosynthesis of Collagen in Human Dermal Fibroblasts (사람 섬유아세포에서 전호(Anthriscus sylvestris Hoffmann)추출물이 콜라겐 생합성에 미치는 영향)

  • Lee, Woo-Jung;Kim, Yong-Kee;Kim, Su-Nam
    • Korean Journal of Plant Resources
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    • v.25 no.2
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    • pp.240-245
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    • 2012
  • In this study, we investigated the boosting effects on collagen biosynthesis of $Anthriscus$ $sylvestris$ ethanol extract (ASE) in human dermal fibroblasts. To obtain more effective fraction and subfraction for collagen biosynthesis, standard solvent partition and open column chromatography were performed. The EtOH extract, solvent fractions, and 8 EtOAc subfractions were tested for their collagen synthesis capacity by [$^3H$]Proline-incorporation and ELISA assay. ASE increased 25% of total collagen synthesis and 27% of procollagen biosynthesis. The total collagen biosynthesis was increased by EtOAc fraction and E6 subfraction to 28% and 50% respectively. Type I procollagens were also upregulated by EtOAc fraction and E6 subfraction to 30% and 47%, each. Taken together, our data suggest that potential anti-aging effect of ASE on skin is via increasing collagen biosynthesis and effective subtraction is E6 subfraction of EtOAc fraction.

Suppressive Effect of Fermented Angelica tenuissima Root Extract against Photoaging: Possible Involvement of Hemeoxygenase-1

  • Park, Yun-A;Lee, Sung Ryul;Lee, Jin Woo;Koo, Hyun Jung;Jang, Seon-A;Yun, Seung-Won;Kim, Hyun Ju;Woo, Jeong Suk;Park, Myung Rye;Kang, Se Chan;Kim, Youn Kyu;Sohn, Eun-Hwa
    • Journal of Microbiology and Biotechnology
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    • v.28 no.8
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    • pp.1391-1400
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    • 2018
  • Angelica tenuissima root has historically been used as a traditional medicine in Korea. Previous studies have identified the anti-melanogenic effects of the extract of A. tenuissima root fermented by Aspergillus oryzae (FAT). This study investigated the protective effects of FAT against ultraviolet light B exposure (UVB; $30mJ/cm^2$) in HaCaT (human keratinocyte) or Hs68 (human foreskin fibroblast) skin cells. FAT treatment was able to stimulate wound healing rate at the basal condition. FAT also favored the maintenance and/or improvement of extracellular matrix impairment caused by UVB irradiation through: 1) upregulation of procollagen Type-1 synthesis and secretion; 2) suppression of MMP-1 and elastase expression. FAT was able to play a role in the attenuation of inflammatory responses caused by UVB irradiation via upregulation of photo-protective hemeoxygease-1 and suppression of proinflammatory cyclooxygenase-2 expression. After further verification of the anti-photoaging potential of FAT, it could be utilized as an effective ingredient in anti-aging and anti-wrinkle cosmetics.

Anti-Wrinkle Effects of Enzymatic Oyster Hydrolysate and Its Fractions on Human Fibroblasts (굴 효소 가수분해물과 분자량 분획물의 섬유아세포에 대한 주름 개선 효과)

  • Kim, Hyeon-Ah;Park, Si-Hyang;Lee, Su-Seon;Choi, Yeung Joon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.11
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    • pp.1645-1652
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    • 2015
  • We investigated the anti-wrinkle efficacy of hydrolysate from oyster protein by Protamex and Neutrase for the purpose of finding materials to assist skin health originating from marine organisms. There were about 7.9% free amino acids in the oyster hydrolysate, and contents of urea, taurine, alanine, and glycine were high. Oyster hydrolysate also showed collagenase inhibitory activity and was not toxic to CCD986sk human fibroblast cells. Yield of the fractions according to the molecular weight of oyster hydrolysate was 40% for less than 1,000 Da and 60.4% for less than 5,000 Da, respectively. Antioxidative effect, procollagen production, and matrix metalloproteinase-1 inhibitory activity were highest in 1,000~3,000 Da fractions. We observed that oyster hydrolysate and its less than 5,000 Da fraction are potential functional compounds for skin health and for improving wrinkles.

EXPRESSIONS OF VASCULAR METASTASIS RELATED FACTORS IN MURINE ORTHOTOPIC TUMOR MODELS OF SALIVARY GLANDS (마우스에서 확립된 타액선 동위종양에서 혈관성 전이관련 인자의 발현)

  • Jang, Jae-Hyun;Kwon, Kwang-Jun;Park, Young-Wook
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.29 no.6
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    • pp.499-508
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    • 2007
  • Background and Purpose: Some subtypes of malignant salivary gland tumors such as adenoid cystic carcinoma (ACC) frequently result in distant metastasis of vascular origin, which are main causes of treatment failure. The reasons for the affinity for vascular metastatic potential are unclear. Therefore, molecular characteristics that influence the dissemination of metastatic tumor cells are important for the design of more effective treatment of salivary ACC. Tumor angiogenesis has been known to be essential for the distant metastasis of malignant cells. So, we determined expressions of vascular metastasis related factors in orthotopic (parotid) murine models of parotid ACC and compared with those in ectopic (subcutis) tumors of athymic mice. Experimental Design: Using specimens from murine parotid (orthotopic, experimental group) and subcutaneous (ectopic, control group) tumors, which have developed via transplantation of tumor cells, originated from human parotid ACC, we performed immunohistochemical assays with anti-vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF, FGF2), matrix metalloproteinase (MMP)-9, and interleukin (IL)-8 antibodies. We also performed immunohistochemical assays with VEGF receptor (VEGFR)-1, VEGFR-2, VEGFR-3, and phosphorylated VEGFR-2. Results: Transplantation of human ACC tumor cell $(5{\times}10^5)$ into the parotid and subcutis successfully resulted in orthotopic (parotid) and ectopic (subcutaneous) tumors in athymic mice. Immunohistochemical staining demonstrated higher expression of major angiogenic factors (VEGF, bFGF, MMP-9) in the orthotopic tumors than in ectopic tumors (P<0.05). But the expression level of angiogenic receptors were same in orthotopic and ectopic tumors of parotid ACC. Conclusion: VEGF, bFGF, and MMP-9 could be a good candidates for antiangiogenic therapy for the contol of vascular metastatic lesions of salivary ACC.

Hepatocyte Growth Factor is the Key Cytokine in Stimulating Potential Stem Cells in the Cord Blood into Hepatic Lineage Cells

  • Ryu, Kyung-Ha;Cho, Su-Jin;Woo, So-Youn;Seoh, Ju-Young;Jung, Yun-Jae;Han, Ho-Seong
    • IMMUNE NETWORK
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    • v.7 no.3
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    • pp.117-123
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    • 2007
  • Background: This study was designed to investigate the role of the hepatocyte growth factor (HGF) with regards to differentiation of somatic stem cells originating from the human umbilical cord blood (UCB) into hepatic lineage cells in vitro culture system. Methods: Mononuclear cells from UCB were cultured with and without HGF based on the fibroblast growth factor (FGF)-1, FGF-2, and stem cell factor. The cultured cells were confirmed by immunofluorescent staining analysis with albumin (ALB), cytokeratin-19 (CK-19), and proliferating cell nuclear antigen (PCNA) MoAb. ALB and CK-18 mRNA were also evaluated by reverse transcription-polymerase chain reaction. In order to observe changes in proliferating capacity with respect to the cultured period, CFSE with affinity to proliferating cells were tagged and later underwent flow cytometry. Results: In the HGF-treated group, cultured cells had a large oval shaped appearance with adherent, but easily detachable characteristics. In the HGF-non treated group, these cells were spindle-shaped with strong adherent characteristics. Expressions of ALB and CK-19 were evident in HGF-treated group compared to non-expression of those in to HGF-non treated group. Dual immunostaining analysis of the ALB producing cells showed presence of PCNA in their nuclei, and ALB and CK-18 mRNA were detected on the 21st day of cultured cells in the HGF-treated group. Conclusion: Our findings suggest that HGF has a pivotal role in differentiating somatic stem cells of human UCB into hepatic lineage cells in vitro.

Effect of red ginseng NaturalGEL on skin aging

  • Kim, Ye Hyang;Park, Hye Rim;Cha, So Yoon;Lee, So Hun;Jo, Jung Wung;Go, Jung Nam;Lee, Kang Hyuk;Lee, Su Yeon;Shin, Song Seok
    • Journal of Ginseng Research
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    • v.44 no.1
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    • pp.115-122
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    • 2020
  • Background: In aged skin, degradation of collagen fibers, which occupy the majority of the extracellular matrix in the dermis, and changes of aquaporin 3 (AQP3) and skin constituents, such as hyaluronic acid and ceramide, cause wrinkles and decrease skin moisturization to contribute to dryness and lower elasticity skin. Red ginseng (RG) is used as a cosmetic and food material and is known to protect from UVB-induced cell death, increase skin hydration, prevent wrinkles, and have an antioxidative effect. But, in general, RG used as a material is the soluble liquid portion in the solvent, and the part that is not soluble in the solvent is discarded. Thus, we made the whole RG into microgranulation and dispersed in water to produce gel form for using entire RG, and it was named red ginseng NaturalGEL (RG NGEL). Methods: RG NGEL was investigated for matrix metalloproteinases inhibitory activity, induction of Type I collagen, AQP3, hyaluronan synthetase 2, serine palmitoyl transferase, ceramide synthase 3, and filaggrin expression and compared with RG water extract. Results: RG NGEL reduced the levels of UV-induced matrix metalloproteinases and increased Type I collagen in human fibroblast cells and upregulated AQP3, hyaluronan synthetase 2, serine palmitoyl transferase, ceramide synthase 3, and filaggrin expressions in human keratinocytes compared with RG water extract. Conclusion: RG NGEL has the potential as an effective reagent for antiaging cosmetics to improve wrinkle formation and skin hydration.

Effects of the Particulate Matter2.5 (PM2.5) on Lipoprotein Metabolism, Uptake and Degradation, and Embryo Toxicity

  • Kim, Jae-Yong;Lee, Eun-Young;Choi, Inho;Kim, Jihoe;Cho, Kyung-Hyun
    • Molecules and Cells
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    • v.38 no.12
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    • pp.1096-1104
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    • 2015
  • Particulate $matter_{2.5}$ ($PM_{2.5}$) is notorious for its strong toxic effects on the cardiovascular, skin, nervous, and reproduction systems. However, the molecular mechanism by which $PM_{2.5}$ aggravates disease progression is poorly understood, especially in a water-soluble state. In the current study, we investigated the putative physiological effects of aqueous $PM_{2.5}$ solution on lipoprotein metabolism. Collected $PM_{2.5}$ from Seoul, Korea was dissolved in water, and the water extract (final 3 and 30 ppm) was treated to human serum lipoproteins, macrophages, and dermal cells. $PM_{2.5}$ extract resulted in degradation and aggregation of high-density lipoprotein (HDL) as well as low-density lipoprotein (LDL); apoA-I in HDL aggregated and apo-B in LDL disappeared. $PM_{2.5}$ treatment (final 30 ppm) also induced cellular uptake of oxidized LDL (oxLDL) into macrophages, especially in the presence of fructose (final 50 mM). Uptake of oxLDL along with production of reactive oxygen species was accelerated by $PM_{2.5}$ solution in a dose-dependent manner. Further, $PM_{2.5}$ solution caused cellular senescence in human dermal fibroblast cells. Microinjection of $PM_{2.5}$ solution into zebrafish embryos induced severe mortality accompanied by impairment of skeletal development. In conclusion, water extract of $PM_{2.5}$ induced oxidative stress as a precursor to cardiovascular toxicity, skin cell senescence, and embryonic toxicity via aggregation and proteolytic degradation of serum lipoproteins.