• Title/Summary/Keyword: Extracellular matrix

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The Effects of Various Extracellular Matrices on Motility of Cultured MC3T3-E1 Cell (다양한 세포외기질이 배양 골아세포의 이동에 미치는 영향)

  • Park, Beyoung Yun;Seo, Sang Woo;Lee, Won Jai;Ryu, Chang Woo;Rah, Dong Kyun;Son, Hyun Joo;Park, Jong Chul
    • Archives of Plastic Surgery
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    • v.32 no.2
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    • pp.143-148
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    • 2005
  • Chemotactic migration of bone forming cell, osteoblast, is an important event during bone formation, bone remodeling, and fracture healing. Migration of cells is mediated by adhesion receptors, such as integrins, that link the cell to extracellular matrix ligands, type I collagen, fibronectin, laminin and depend on interaction between integrin and extracellular ligand. Our study was designed to investigate the effect of extracellular matrix like fibronectin, laminin, type I collagen on migration of osteoblast. Migration distance and speed of MC3T3-E1 cell on extracellular matrix-coated glass were measured for 24 hours using 0.01% type I collagen, 0.01% fibronectin, 100 microliter/ml laminin. The migration distance and speed of MC3T3-E1 cell was compared using a video-microscopy system. To determine migration speed, cells were viewed with a 4 phase- contrast lens and video recorded. Images were captured using a color CCD camera and saved in 8-bit full-color mode. The migration distance on 0.01% type I collagen or 0.01% fibronectin was longer than that on $100{\mu}l/ml$ laminin-coated glass. The migration speed on fibronectin-coated glass was 68 micrometer/hour which was fastest. The migration speed on type I collagen-coated glass was similar with that on fibronectin-coated glass. The latter two migration speeds were faster than that on no-coated glass. On the other hand, the average migration speed on laminin-coated glass was 37micrometer/hour and not different from that of control group. In conclusion, the extracelluar matrix ligands such as type I collagen and fibronectin seem to play an important role in cell migration. The type I collagen or fibronectin coated scaffold is more effective for migration of osteoblast in tissue engineering process.

Cloning, Purification, and Characterization of Recombinant Human Extracellular Superoxide Dismutase in SF9 Insect Cells

  • Shrestha, Pravesh;Yun, Ji-Hye;Kim, Woo Taek;Kim, Tae-Yoon;Lee, Weontae
    • Molecules and Cells
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    • v.39 no.3
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    • pp.242-249
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    • 2016
  • A balance between production and degradation of reactive oxygen species (ROS) is critical for maintaining cellular homeostasis. Increased levels of ROS during oxidative stress are associated with disease conditions. Antioxidant enzymes, such as extracellular superoxide dismutase (EC-SOD), in the extracellular matrix (ECM) neutralize the toxicity of superoxide. Recent studies have emphasized the importance of EC-SOD in protecting the brain, lungs, and other tissues from oxidative stress. Therefore, EC-SOD would be an excellent therapeutic drug for treatment of diseases caused by oxidative stress. We cloned both the full length (residues 1-240) and truncated (residues 19-240) forms of human EC-SOD (hEC-SOD) into the donor plasmid pFastBacHTb. After transposition, the bacmid was transfected into the Sf9-baculovirus expression system and the expressed hEC-SOD purified using FLAG-tag. Western blot analysis revealed that hEC-SOD is present both as a monomer (33 kDa) and a dimer (66 kDa), as detected by the FLAG antibody. A water-soluble tetrazolium (WST-1) assay showed that both full length and truncated hEC-SOD proteins were enzymatically active. We showed that a potent superoxide dismutase inhibitor, diethyldithiocarbamate (DDC), inhibits hEC-SOD activity.

Effect of Extracellular Matrix Proteins on the In Vitro Development of Parthenogenetic Mouse Eggs (세포외 기질 단백질이 생쥐 단위발생란의 체외 발달에 미치는 영향)

  • 곽대오;김선구;김영수;박충생
    • Journal of Embryo Transfer
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    • v.8 no.2
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    • pp.83-90
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    • 1993
  • To investigate the effect of extracellular matrix proteins on the in vitro development of ethanol-induced parthenogenetic eggs of ICR strain mice, those were cultured in vitro in fibronectin, gelatin, or collagen precoated culture dishes containing 1.5 ml of NaH-C03$_3$-BMOC-3 medium at 37$^{\circ}C$ for 96 hrs. under the atmosphere of 5% $CO_2$ and 95% air. Fibronectin, gelatin, or collagen significantly(P$\pm$1.4, 45.4i1.4, and 44.8$\pm$O.9, respectively. And the diameter of those eggs ranged 104.6$\pm$1.9, 102.8$\pm$2.3, and 103.4$\pm$O.8 $\mu$m, respectively.

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Extracellular matrixes and neuroinflammation

  • Jang, Dong Gil;Sim, Hyo Jung;Song, Eun Kyung;Kwon, Taejoon;Park, Tae Joo
    • BMB Reports
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    • v.53 no.10
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    • pp.491-499
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    • 2020
  • The extracellular matrix is a critical component of every human tissue. ECM not only functions as a structural component but also regulates a variety of cellular processes such as cell migration, differentiation, proliferation, and cell death. In addition, current studies suggest that ECM is critical for the pathophysiology of various human diseases. ECM is composed of diverse components including several proteins and polysaccharide chains such as chondroitin sulfate, heparan sulfate, and hyaluronic acid. Each component of ECM exerts its own functions in cellular and pathophysiological processes. One of the interesting recent findings is that ECM is involved in inflammatory responses in various human tissues. In this review, we summarized the known functions of ECM in neuroinflammation after acute injury and chronic inflammatory diseases of the central nerve systems.

Shotgun Phage Display of Lactobacillus casei BL23 Against Collagen and Fibronectin

  • Munoz-Provencio, Diego;Monedero, Vicente
    • Journal of Microbiology and Biotechnology
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    • v.21 no.2
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    • pp.197-203
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    • 2011
  • Lactobacilli are normal constituents of the intestinal microbiota, and some strains show the capacity to bind to extracellular matrix proteins and components of the mucosal layer, which represents an adaptation to persist in this niche. A shotgun phage-display library of Lactobacillus casei BL23 was constructed and screened for peptides able to bind to fibronectin and collagen. Clones showing binding to these proteins were isolated, which encoded overlapping fragments of a putative transcriptional regulator (LCABL_29260), a hypothetical protein exclusively found in the L. casei/rhamnosus group (LCABL_01820), and a putative phage-related endolysin (LCABL_13470). The construction of different glutathione S-transferase (GST) fusions confirmed the binding activity and demonstrated that the three identified proteins could interact with fibronectin, fibrinogen, and collagen. The results illustrate the utility of phage display for the isolation of putative adhesins in lactobacilli. However, it remains to be determined whether the primary function of these proteins actually is adhesion to mucosal surfaces.

Potential Roles of Protease Inhibitors in Cancer Progression

  • Yang, Peng;Li, Zhuo-Yu;Li, Han-Qing
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8047-8052
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    • 2016
  • Proteases are important molecules that are involved in many key physiological processes. Protease signaling pathways are strictly controlled, and disorders in protease activity can result in pathological changes such as cardiovascular and inflammatory diseases, cancer and neurological disorders. Many proteases have been associated with increasing tumor metastasis in various human cancers, suggesting important functional roles in the metastatic process because of their ability to degrade the extracellular matrix barrier. Proteases are also capable of cleaving non-extracellular matrix molecules. Inhibitors of proteases to some extent can reduce invasion and metastasis of cancer cells, and slow down cancer progression. In this review, we focus on the role of a few proteases and their inhibitors in tumors as a basis for cancer prognostication and therapy.

Amine Oxidase Activity of the Human Lysyl Oxidase-Like 3 (LOXL3) Protein

  • Kim Young-Ho
    • Biomedical Science Letters
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    • v.12 no.3
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    • pp.185-190
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    • 2006
  • Lysyl oxidase (LOX) catalyzes the lysine-derived cross-links of fibrillar collagens and elastin in the extracellular matrix. Recent molecular cloning has revealed existence of a LOX family consisting of LOX and four lysyl oxidase-like proteins (LOXL, LOXL2, LOXL3 and LOXL4). Pathological conditions associated with impaired LOX activity in several heritable and acquired disorders lead to severe structural and functional abnormalities of cardiovascular tissues, such as occlusion of coronary arteries and aneurysms, suggesting an essential role for the LOX family proteins in the maintenance of the cardiovascular system. However, the specific roles of the lysyl oxidase-like proteins in normal and pathological conditions of the cardiovascular tissues have not been established yet. Here, I report that LOXL3, a novel member of the LOX family, is predominantly expressed in the aorta, with an amine oxidase activity toward collagen and elastin, suggesting an essential role of LOXL3 in the development and maintenance of the aorta.

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Mechanical/Biochemical Analysis of Cell Adhesion Strengthening (세포흡착 거동의 기계적/생화학적 분석)

  • Shin, Heung-Soo
    • Proceedings of the KSME Conference
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    • 2008.11a
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    • pp.1455-1457
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    • 2008
  • Cell adhesion is a coordinated process involving initial binding of integrin receptors to extracellular matrix (ECM), recruitment of adhesion proteins, and focal adhesion assembly. The formation of mechanically stable focal adhesion assembly of cells within surrounding ECM is a key parameter to direct numerous cellular functions including cell migration, differentiation, and apotosis. With current cell adhesion assays, it is difficult to understand contributions of each coordinated event on evolution of cell adhesion strengthening since cells spontaneously spread upon their adhesion to the substrate, thus remodeling their cytoskeletal structure. In this presentation, novel approaches for analysis of cell adhesion strengthening process based on the combination of mechanical device, micro-patterned substrates, and molecular biological techniques will be discussed.

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Stem cell maintenance in a different niche

  • Lim, Jeong Mook;Ahn, Ji Yeon;Lee, Seung Tae
    • Clinical and Experimental Reproductive Medicine
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    • v.40 no.2
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    • pp.47-54
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    • 2013
  • To overcome the difficulty of controlling stem cell fate and function in applications to regenerative medicine, a number of alternative approaches have been made. Recent reports demonstrate that a non-cellular niche modulating the biophysical microenvironment with chemical factors can support stem cell self-renewal. In our previous studies, early establishment was executed to optimize biophysical factors and it was subsequently found that the microgeometry of the extracellular matrix made huge differences in stem cell behavior and phenotype. We review here a three-dimensional, non-cellular niche designed to support stem cell self-renewal. The characteristics of stem cells under the designed system are further discussed.