• Title/Summary/Keyword: fibronectin

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A Role of Fibronectin in the Extracellular Matrix during Chick Mvoblast Differentiation (계배 근원세포의 분화에서 Extracellular matrix내 fibronectin의 역할)

  • 문경엽;하두봉정진하강만식
    • The Korean Journal of Zoology
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    • v.38 no.1
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    • pp.78-86
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    • 1995
  • Our previous report has suggested that the decrease of fibronectin level during mvogenesis is due to the decreased Bvailabilitv of receptor (matrix assembly receptor) for 29-kDa fragment of fibronectin. In the present study, we demonstrate that G protein and adenvlate cvclase system are involved in the regulation of fibronectin matrix assembly and that when fibronectin level in extracellular matrix decreases, the postmitotic fusion-capable cells emerge more frequently from the proliferative population. This proposal is based on the following observations. (1) Cholers toxin, which increases intracellular CAMP, caused a decrease in the ability of mvoblasts to incorporate fibronectin into extracellular matrix. (2) Cholera toxin decreased the proliferation of mvoblasts and Induced the precocious fusion. (3) decAMP, which was found to induce the precocious fusion and decrease the proliferation of myoblasts, decreased the fibronectin level in extracellular matrix and matrix assembly receptor for fibronectin- (4) RGOS, whlh inhibits the incorporation of fibronectin into extracellular matrix, induced the precocious fusion and reduced the proliferaton of mvoblasts. These results suggest that CAMP regulates the fibronectin levels in extracellular matrix and that the alteration of fibronectin level is involved in regulation of the proliferation and differentiation of chick embryonic mvoblasts.

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Alterations in the Level of Fibronectin and its Receptors during Chick Myoblast Differentiation (계배 근원세포 분화에 따른 Fibronection의 수준과 그 수용체의 변화)

  • 정창용;강만식
    • The Korean Journal of Zoology
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    • v.31 no.2
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    • pp.95-103
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    • 1988
  • Alterations in the amount of fibronectin during chick myogenesis were investigated. The amount of fibronectin as measured by immunoblotting was found to decrease during the process. As a first step in answering the precise mode of change in the level of ibronedin during myogensis, the interaction of 28,000 dalton(28 kDa) amino terminal fragment of fibronectin as well as 85,000 dalton (85 kDa) fragrxient with myoblasts was examined. The specific binding of 125 l-28 kDa fragment to myoblasts was time-dependent and reached a maximum within 60 min. Unlabelled 28 kDa fragment inhibited the binding of 125 I-28 kDa fragment, whereas 85 kDa fragment containing adhesion promoting activity did not inhibit it. This finding suggests that the 28 kDa fragment interacts with the matrix assembly receptors but not with the cell adhesion receptors. Accordingly, the decrease in the level of fibronectin is likely to correlate with the fall of fibronectin receptors on the myoblasts.

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The fibronectin concentration that optimally maintains porcine satellite cells

  • Jae Ho Han;Si Won Jang;Ye Rim Kim;Hoon Jang;Kwan Seob Shim;Hyun Woo Choi
    • Animal Bioscience
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    • v.36 no.12
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    • pp.1889-1897
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    • 2023
  • Objective: 'Cultured meat' has been suggested as means of solving the problems associated with overpopulation and gas emissions. Satellite cells are a major component in the production of cultured meat; however, these cells cannot be maintained in vitro over long periods. Fibronectin is a glycoprotein that affects biological processes such as cell adhesion, differentiation, and migration. Unfortunately, the characteristics of porcine satellite cells grown in a long-term culture when exposed to fibronectin-coated dishes are unknown. The objective of this study was to investigate the appropriate concentration of fibronectin coated dishes for proliferation and maintenance of porcine satellite cells at long-term culture. Methods: In this study, we isolated the satellite cells and fibroblast cells with pre-plating method. We next analyzed the cell doubling time, cell cycle, and rate of expressed paired box 7 (Pax7) and myogenic differentiation 1 (MyoD1) in porcine satellite cells cultured with 20 ㎍/mL of fibronectin-, gelatin-, and non-coated dishes at early and late passage. We then analyzed the proliferation of porcine satellite cells with various concentrations of mixed gelatin/fibronectin. We next determined the optimal concentration of fibronectin that would encourage proliferation and maintenance of porcine satellite cells in a long-term culture. Results: Doubling time was lowest when 20 ㎍/mL of fibronectin was used (as tested during an early and late passage). Levels of expressed Pax7 and MyoD1, assessed using immunocytochemistry, were highest in cells grown using fibronectin-coated dishes. The proliferation of gelatin/fibronectin mixed coatings had no significant effect on porcine satellite cells. The concentration of 5 ㎍/mL fibronectin coated dishes showed the lowest doubling time and maintained expression of Pax7. Conclusion: Fibronectin with 5㎍/mL effectively maintains porcine satellite cells, a discovery that will be of interest to those developing the next generation of artificial meats.

Biological Effects of Fibronectin Type III 10 domain on Human Osteoblast-like cells (Fibronectin type III 10 도메인이 조골양 세포에 미치는 생물학적 영향)

  • Lee, Chang-Seok;Jang, Jun-Hyeog;Kim, Tae-Il;Lee, Yong-Moo;Rhyu, In-Chul;Chung, Chong-Pyoung;Han, Soo-Boo;Ku, Young
    • Journal of Periodontal and Implant Science
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    • v.34 no.2
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    • pp.293-301
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    • 2004
  • 1. 연구 목적 Fibronectin은 세포외기질의 주요성분인 거대 당단백질로서, 조골세포의 부착과 증식 및 이동능에 중요한 역할을 담당한다고 알려져 있다. 이러한 fibronectin의 조골세포에 대한 영향을 실제 임상에 적용하기 위해서, 전체 fibronectin 단백질을 사용하는 것은 면역학적으로나 경제적으로 많은 단점을 안고 있어서, 유효한 반응단위만을 추출하여 활용하는 것이 바람직한 방법으로 알려져 있다. 이 연구의 목적은 세포부착에 주로 관여하는 fibronectin type III분절 중 10번 도메인이 조골양 세포에 미치는 영향을 전체 fibronectin단백질과 fibronectin type III 7-10 도메인 분절과 비교, 관찰하는 것이다. 2. 연구 방법 사람의 fibronectin을 기초로 한 적절한 primer로서, 유전자 재조합법을 이용하여 fibronectin type III 10 도메인과 fibronectin type III 7-10 도메인 분절을 얻었으며, 전체 fibronectin분자는 상용으로 준비하여 24-well 세포배양 용기에 도포하였다. 배양된 조골양세포(HOS cell)를 $1x10^5$ cells/well의 농도로 각 well에 분주하여 $37^{\circ}C$에서 1시간 배양을 하였다. Cell adhesion assay를 실시하기 위해 10% formaldehyde로 고정시키고 1% Crystal Violet으로 염색하여 광학현미경을 관찰 후 2% SDS를 처리하여 microplate reader기를 이용하여 570nm에서 혼탁정도를 측정하였다. 음성대조군으로는 RPMI 용액을 사용하였다. 동일한 방법을 이용하여 준비된 $35mm^2배양접시에 HOS cell을 $37^{\circ}C$에서 4일간 배양 후, MTS assay를 이용하여 세포 증식도에 미치는 영향을 관찰하였다. 6일째 405nm에서 활성화된 세포에서 분비된 p-nitrophenol을 이용한 alkaline phosphatase activity를 측정하였다. 3. 결과 및 고찰 Fibronectin type III 10 도메인은 HOS cell에 대한 생물학적인 효과면에서, 전체 fibronectin 분자 및 fibronectin type III 7-10 분절과 통계적으로 유사한 세포부착도를 보여주었으며, 세포증식도와 alkaline phosphatse 활성도면에서도 큰 차이가 나타나지 않았다. 이상의 연구결과로 볼 때, fibronectin type III 10 도메인이 조골세포의 증식을 목적으로 사용하는 생체재료의 표면개질 부착물질로 응용할 수 있는 가능성이 있다고 하겠다.

Fibronectin Induces Pro-MMP-2 Activation and Enhances Invasion in H-Ras-Transformed Human Breast Epithelial Cells

  • Kim, Jong-Sook;Moon, A-Ree
    • Biomolecules & Therapeutics
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    • v.17 no.3
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    • pp.288-292
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    • 2009
  • Interactions between tumor cells and the extracellular matrix (ECM) strongly influence tumor development, affecting cell survival, proliferation and migration. Fibronectin, a major component of ECM, has been shown to interact with integrins especially the ${\alpha}5{\beta}1$ integrin. Cell invasion and metastasis are often associated with matrix metalloproteinases (MMPs) which are capable of digesting the different components of the ECM and basement membrane. MMP-2 is produced as a latent pro-MMP-2 (72 kDa) to be activated, resulting the 62 kDa active MMP-2. In this study, we investigated the effect of fibronectin on activation of pro-MMP-2 and the cellular invasiveness in H-Ras-transformed MCF10A human breast epithelial cells. Here we show that fibronectin induces activation of pro-MMP-2 and up-regulation of MT1-MMP and TIMP-2 in H-Ras MCF10A cells. These results demonstrate that H-Ras MCF10A cells secrete high levels of active MMP-2 when cultured with fibronectin, suggesting a possible interaction between the ECM network and H-Ras MCF10A cells to generate active MMP-2 which is important for proteolysis and ECM remodeling. Invasive and migratory abilities of H-Ras MCF10A cells were enhanced by fibronectin. Fibronectin up-regulated the expression of ${\beta}1$ integrin which may play a role in cellular responses exerted by fibronectin. Since acquisition of pro-MMP-2 activation can be associated with increased malignant progression, this study provides a mechanism for the cell surface-matrix degrading effect of fibronectin which will be crucial to breast cell invasion and migration.

The Experimental Studies Of The Kaejibokryungwhan, Sobokchugeotang And Dohongsamultang On The Mesangial Cell Proliferation And Fibronectin Synthesis (계지복령환(桂枝茯笭丸), 소복축어탕(少腹逐瘀湯), 도홍사물탕(桃紅四物湯)이 메산지움세포 증식과 Fibronectin 합성에 미치는 영향(影響))

  • Ahn Young-Min;Ahn Se-Young;Doo Ho-Kyung;Kwon Young-Ku
    • The Journal of Internal Korean Medicine
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    • v.24 no.1
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    • pp.94-103
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    • 2003
  • In order to investigate the effects of Kaejibokryungwhan, Sobokchugeotang and Dohongsamultang on mesangial cell proliferation and fibronectin synthesis, laboratory study was performed. The results are summarized as follows; 1. Mesangial cell proliferation was significantly decreased in the Kaejibokryungwhan group and the Dohongsamultang group compared with the Control group. In the Sobokchugeotang group, the mesangia1 cell proliferation activity was lesser than Control group, but it was statistically non-significant. In Kaejibokryungwhan group, the Sobokchugeotang group and Dohongsamultang group, mesangial cell proliferation was significantly decreased compared with the Hydrocortisone group 2. In the group, which contained fetal bovine serum, fibronectin synthesis was decreased in the Kaejibokryungwhan group, the Sobokchugeotang group and Dohongsamultang group compared with Control group, but the difference was statistically non-significant. In Kaejibokryungwhan group, Sobokchugeotang group and Dohongsamultang group, fibronectin was less decreased compared with that of Hydrocortisone group. 3. In the group, which contained fetal bovine serum, fibronectin synthesis was significantly decreased in Kaejibokryungwhan group and Sobokchugeotang group than those of Control group. In Dohongsamultang group, the fibronectin synthesis was decreased than Control group, but the difference was statistically non-significant. In Sobokchugeotang group, the fibronectin synthesis was decreased than Hydrocortisone group, but it was statistically non-significant. According to the above results, the mesangial cell proliferation and fibronectin systhesis could be reduced by Kaejibokryungwhan group significantly.

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The Chemokine SDF-1α Suppresses Fibronectin-mediated In Vitro Lymphocytes Adhesion

  • Ji, LiLi;Sheng, YuChen;Wang, ZhengTao
    • Molecules and Cells
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    • v.22 no.3
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    • pp.308-313
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    • 2006
  • Stromal cell-derived factor (SDF-1) is a CXC chemokine that selectively activates the CXCR4 chemokine receptor. Fibronectin is an intracellular matrix component that binds integrin and mediates cell-matrix adhesion. Activation of the integrin receptor can occur in two ways: by ligand binding (outside-in signaling), and in response to intracellular events (inside-out signaling). In the current study we showed that SDF-$1{\alpha}$ inhibited adhesion of T lymphocyte Jurkat cells resulting from binding high concentrations of fibronectin as well as that of THP-1 monocytes. The effect of SDF-$1{\alpha}$ on fibronectin-mediated adhesion was partly reversed by the CXCR4 receptor antagonist T140. Our results suggest that an SDF-1/CXCR4 signal pathway modulates fibronectin-mediated lymphocytes adhesion.

Immune Response of Alpha-toxin, Capsular Polysaccharide (CPS) and Recombinant Fibronectin-Binding Protein (r-FnBP) of Staphylococcus aureus in Rabbit (황색포도상구균의 Alpha-toxin, Capsular Polysaccharide (CPS)와 재조합 Fibronectin-Binding Protein (r-FnBP) 항원을 이용한 토끼에서의 면역반응)

  • Park, Hee-Myung;Yoo, Han-Sang;Han, Hong-Ryul;Kim, Doo
    • Journal of Veterinary Clinics
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    • v.15 no.2
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    • pp.370-377
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    • 1998
  • 본 연구는 황색포도상구균의 병원성 인자인 alpha-toxin, casular polysaccharides (CPS)와 fibronectin-binding protein (FnBP)을 이용해 실험동물인 토끼에서의 항체가 형성 능과 공격접종 후 방어능에 관한 연구를 위하여 수행되었으며 향후 젖소 유방염 아단위 항원 을 이용한 백신개발 가능성을 탐색하고자 수행되어졌다. 황색포도상구균의 alpha-toxin, capsu18r polysaccharides (CPS)91 fibronectin-binding protein (FnBP) 항원을 이통재 효소면 역측정법을 통한 혈중 IgG 항체가수준을 측정하였으며 alpha-toxin, capsular polysaccharides (CPS)와 fibronectin-binding protein (FnBP)으로 면역시킨 토끼에서 대조군의 토끼보다 혈 중 항체가 수준이 1차 면역 이후 유의성 있게 높았다 (p<0.05). 백신에 사용된 alpha-toxin, capsular polysaccharides (CPS)와 fibronectin-binding protein (FnBP) 항원중 capsular polysaccharides (CPS)가 다른 alpha toxin과 fibronectin-binding protein (FnBP)의 혈중항 체가 수준과 비교하여 볼 때 비교적 낮은 수준이었다. 세균을 혈중으로 공격접종한 후 대조군 과 백신접종군의 혈중내 세균제거율에 있어 대조군에 비해 백신접종군에서 유의성 있게 낮은 균수를 보였다 (p<0.05). 또한 장기내 균수측정실험결과 대조군의 장기보다 백신접종군에서 유의성 있게 세균수가 낮게 출현하였다 (p<0.05). 결론적으로 본 연구결과 황색포도상구균의 3가지 항훤 alpha-toxin, capsular polysaccharide와 재조합 fibronectin binding protein을 이 퐁한 실험동물에서 안단위 유방염 백신은 방어능이 있다고 생각된다.

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EFFECTS OF VARIOUS SURFACE TREATMENTS FOR TITANIUM ON SURFACE MICRO ROUGHNESS, STATIC WETTABILITY, FIBRONECTIN ADSORPTION (표면 처리 방법에 따른 타이타늄의 미세 표면 거칠기, 표면 젖음성, fibronectin 흡착량에 미치는 영향)

  • Shin Hwa-Sub;Kim Young-Su;Shin Sang-Wan
    • The Journal of Korean Academy of Prosthodontics
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    • v.44 no.4
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    • pp.443-454
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    • 2006
  • Purpose: This study aims to get the fundamental data which is necessary to the development direction of implant surface treatment hereafter, based on the understanding the surface structure and properties of titanium which is suitable for the absorption of initial tissue fluid by researching effects of additional surface treatments fir sandblasted with large git and acid-etched(SLA) titanium on surface micro-roughness, static wettability, fibronectin adsorption Materials and Method: In the Control groups, the commercial pure titanium disks which is 10mm in diameter and 2mm in thickness were treated with HCI after sandblasting with 50$\mu$m $Al_2O_3$. The experiment groups were made an experiment each by being treated with 1) 22.5% nitric acid according to SLA+ASTM F86 protocol, 2) SLA+30% peroxide, 3) SLA+NaOH, 4) SLA+ Oxalic acid, and 5) SLA+600$^{\circ}C$ heating. In each group, the value of Ra and RMS which are the gauges of surface roughness was measured, surface wettability was measured by analyzing with Sessile drop method, and fibronectin adsorption was measured with immunological assay. The significance of each group was verified by (SPSS, ver.10.0 SPSS Inc.) Kruskal-Wallis Test. (α=0.05) And the correlation significance between Surface micro-roughness and surface wettability. surface roughness and fibronectin adsorption, and surface wettability and fibronectin adsorption was tested by Spearman's correlation analysis. Result: All measure groups showed the significant differences in surface micro-roughness, surface wettability, and fibronectin adsorption. (p<0.05) There was no significance in correlation among the surface micro-roughness, surface wettability, and fibronectin adsorption. (p>0.05) Conclusion: Surface micro-roughness and surface wettability rarely affected the absorption of initial tissue fluid on the surface of titanium.

AN IMMUNOHISTOCHEMICAL STUDY OF THE DISTRIBUTION OF FIBRONECTIN, LAMININ AND TENASCIN IN THE REGENERATING PERIODONTAL TISSUE (재생중인 치주조직내 Fibronectin, Laminin 및 Tensacin의 분포에 관한 면역조직화학적 연구)

  • Chung, Gap-Hwan;Kim, Byung-Ok;Han, Kyung-Yoon
    • Journal of Periodontal and Implant Science
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    • v.25 no.2
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    • pp.321-340
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    • 1995
  • The regeneration of destructed periodontal tissues is one of the ultimate objectives of periodontal therapy. Guided tissue regeneration technique was developed for the ideal regeneration of periodontal tissues. In order to investigate the role of fibronectin, laminin and tenascin in the regenerating process of periodontal tissues, the expanded PTFE barrier membranes(Gore Associates, USA) removed from the patients who had been treated by guided tissue regeneration(GTR) and guided bone regeneration(GBR) techniques were fixed in neutral formalin for 6-24 hours, embedded with paraffin, sectioned at $4-6{\mu}m$ in thickness, and immunohistochemically processed by Avidin-Biotin peroxidase complex method for detecting fibronectin, laminin and tenascin. Monoclonal mouse anti-human fibronectin antibody(Oncogene Science, USA., 1:100), monoclonal mouse anti-human laminin antibody(Oncogene Science, USA., 1:50) and mouse anti-human tenascin antibody(Oncogene Science, USA, 1:10) were used as primary antibodies. The light microscopic findings were as follows: (1) The distribution of fibronectin, laminin and tenascin was various according to the area of barrier membranes. (2) The distribution of fibronectin in case of GBR was extensive in the tissue on the outer surface of barrier membranes, and rare in the intervening space and on the inner surface. In case of GTR it was extensive on the outer surface and in the intervening space, and rare on the inner surface. (3) The distribution of laminin was rare in the tissue on the outer, the inner surface and intervening space of barrier membranes, regardless of GBR or GTR. (4) In case 'of GBR rare distribution of tenascin was observed on the outer surface only, except the inner surface and the intervening space of barrier membranes. In case of GTR the distribution of tenascin was extensive in the tissue on the outer surface, rare in intervening space and the inner surface. The results suggest that fibronectin, laminin and tenascin may play a important role in the regenerating process of periodontal tissue, and they may affect the outcome of healing.

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