• Title/Summary/Keyword: cell surface protein

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Interaction of Different Types of Cells on Poly(L-lactide-co-glycolide) Surface with Wettability Chemogradient

  • Gilson Khang;John M. Rhee;Lee, Jin-Ho;Lee, Ilwoo;Lee, Hai-Bang
    • Macromolecular Research
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    • v.8 no.6
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    • pp.276-284
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    • 2000
  • A wettability chemogradient on poly(L-lactide-co-glycolide) (PLGA) films was prepared by treating the films in air with corona from a knife-type electrode whose power increases gradually along the sample length. The PLGA surfaces oxidized gradually with the increasing corona power, and the wettability chemogradient was created on the surfaces as evidenced by the measurement of water contact angles and electron spectroscopy for chemical analysis. The wettability chemogradient PLGA surfaces were used to investigate the interaction of four different types of cells such as hepatoma (Hep G2), osteoblast (MG 63), bovine aortic endothelial (CPAE), and fibroblast (NIH/3T3) cells in terms of the surface hydrophilicity/hydrophobicity of PLGA. The cells adhered and grown on the chemogradient surface along the sample length were counted and observed by scanning electron microscopy. It was observed that the cells were adhered, spread, and grown more onto the positions with moderate hydrophilicity of the wettability chemogradient PLGA surface than the more hydrophobic or hydrophillic positions, regardless of the cell types used. The maximum adhesion and growth of the cells appeared at around water contact angles of 53~55°. This result seems closely related with the serum protein adsorption on the surface; the serum proteins were also adsorbed more onto the positions with moderate hydrophilicity of the wettability chemogradient surface. It seems that the wettability plays important roles for cell adhesion, spreading and growth on the PLGA surface. The surface modification technique used in this study may be applicable tothe area of tissue engineering for the improvement of tissue compatibility of films- or scaffold-type substrates.

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The Presence in Embryo Extract of a Myotrophic Protein That Affects Proliferation and Fusion of Chick Embryonic Myoblasts in Culture (배양 계배 근원세포의 분화에 미치는 계배 추출물내 Myotrophic Protein의 영향)

  • 유병제;이창호;곽규봉;정진하;하두봉
    • The Korean Journal of Zoology
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    • v.31 no.3
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    • pp.207-217
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    • 1988
  • A myotrophic protein that seemed to he eseentiai for the hision of chick embryonic myoblasts in culture was isolated from chick embryo extrad and was found to be identical or at least similar to the iron-transporting protein, transferrin. Embryo extract seemed to contain, in addition to this myotrophic protein, a heat stable protein that inhibits the fusion of myoblasts. Iron seemed to he necessary for myoblasts to fuse and it was supposed that the role of the myotrophic protein m myoblast fusion is to supply iron to the cell. The numher of the myotrophic protein receptors on myoblast surface membrane decreased immediately after the start of myoblast fusion, supposedly due to the decreased need of iron after the fusion once commenced. It was estimated that endocytosis of myotrophic protein took about 10 minutes and one recycling about 2 hours. The accumulation of iron in myoblasts continued linearly with cultre time and endocytosis of the myotrophic protein occured at a constant rate.

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Comparative Analysis of Envelope Proteomes in Escherichia coli B and K-12 Strains

  • Han, Mee-Jung;Lee, Sang-Yup;Hong, Soon-Ho
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.470-478
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    • 2012
  • Recent genome comparisons of E. coli B and K-12 strains have indicated that the makeup of the cell envelopes in these two strains is quite different. Therefore, we analyzed and compared the envelope proteomes of E. coli BL21(DE3) and MG1655. A total of 165 protein spots, including 62 nonredundant proteins, were unambiguously identified by two-dimensional gel electrophoresis and mass spectrometry. Of these, 43 proteins were conserved between the two strains, whereas 4 and 16 strain-specific proteins were identified only in E. coli BL21(DE3) and MG1655, respectively. Additionally, 24 proteins showed more than 2-fold differences in intensities between the B and K-12 strains. The reference envelope proteome maps showed that E. coli envelope mainly contained channel proteins and lipoproteins. Interesting proteomic observations between the two strains were as follows: (i) B produced more OmpF porin with a larger pore size than K-12, indicating an increase in the membrane permeability; (ii) B produced higher amounts of lipoproteins, which facilitates the assembly of outer membrane ${\beta}$-barrel proteins; and (iii) motility- (FliC) and chemotaxis-related proteins (CheA and CheW) were detected only in K-12, which showed that E. coli B is restricted with regard to migration under unfavorable conditions. These differences may influence the permeability and integrity of the cell envelope, showing that E. coli B may be more susceptible than K-12 to certain stress conditions. Thus, these findings suggest that E. coli K-12 and its derivatives will be more favorable strains in certain biotechnological applications, such as cell surface display or membrane engineering studies.

Alteration of Apoptosis during Differentiation in Human Dental Pulp-Derived Mesenchymal Stem Cell

  • Lee, Hyeon-Jeong;Park, Byung-Joon;Jeon, Ryoung-Hoon;Jang, Si-Jung;Son, Young-Bum;Lee, Sung-Lim;Rho, Gyu-Jin;Kim, Seung-Joon;Lee, Won-Jae
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.1
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    • pp.2-9
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    • 2019
  • Because mesenchymal stem cells (MSCs) maintain distinct capacities with respect to self-renewal, differentiation ability and immunomodulatory function, they have been highly considered as the therapeutic agents for cell-based clinical application. Of particular, differentiation condition alters characteristics of MSCs, including cellular morphology, expression of gene/protein and cell surface molecule, immunological property and apoptosis. However, the previous results for differentiation-related apoptosis in MSCs have still remained controversial due to varied outcomes. Therefore, the present study aimed to disclose periodical alterations of pro- and anti-apoptosis in MSCs under differentiation inductions. The human dental pulp-derived MSCs (DP-MSCs) were differentiated into adipocytes and osteoblasts during early (1 week), middle (2 weeks) and late (3 weeks) stages, and were investigated on their apoptosis-related changes by Annexin V assay, qRT-PCR and western blotting. The ratio of apoptotic cell population was significantly (p < 0.05) elevated during the early to middle stages of differentiations but recovered up to the similar level of undifferentiated state at the late stage of differentiation. In the expression of mRNA and protein, whereas expressions of pro-apoptosis-related makers (BAX and BAK) were not altered in any kind and duration of differentiation inductions, anti-apoptosis marker (BCL2) was significantly (p < 0.05) elevated even at the early stage of differentiations. The recovery of apoptotic cell population at the late stage of differentiation is expected to be associated with the response by elevation of anti-apoptotic molecules. The present study may contribute on understanding for cellular mechanism in differentiation of MSCs and provide background data in clinical application of MSCs in the animal biotechnology to develop effective and safe therapeutic strategy.

Porcine Knock-in Fibroblasts Expressing hDAF on α-1,3-Galactosyltransferase (GGTA1) Gene Locus

  • Kim, Ji-Woo;Kim, Hye-Min;Lee, Sang-Mi;Kang, Man-Jong
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.10
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    • pp.1473-1480
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    • 2012
  • The Galactose-${\alpha}1$,3-galactose (${\alpha}1$,3Gal) epitope is responsible for hyperacute rejection in pig-to-human xenotransplantation. Human decay-accelerating factor (hDAF) is a cell surface regulatory protein that serves as a complement inhibitor to protect self cells from complement attack. The generation of ${\alpha}1$,3-galactosyltransferase (GGTA1) knock-out pigs expressing DAF is a necessary step for their use as organ donors for humans. In this study, we established GGTA1 knock-out cell lines expressing DAF from pig ear fibroblasts for somatic cell nuclear transfer. hDAF expression was detected in hDAF knock-in heterozygous cells, but not in normal pig cells. Expression of the GGTA1 gene was lower in the knock-in heterozygous cell line compared to the normal pig cell. Knock-in heterozygous cells afforded more effective protection against cytotoxicity with human serum than with GGTA1 knock-out heterozygous and control cells. These cell lines may be used in the production of GGTA1 knock-out and DAF expression pigs for xenotransplantation.

Crystal structure of the pretense domain of an ATP-independent heat shock protease HtrA

  • Kim, Dong-Young;Kim, Dong-Ryoung;Ha, Sung-Chul;Neratur K.Lokanath;Hwang, Hye-Yeon;Kim, Kyeong-Kyu
    • Proceedings of the Korea Crystallographic Association Conference
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    • 2002.11a
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    • pp.24-24
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    • 2002
  • HtrA (high temperature requirement A), a periplasmic heat shock protein, is known to have molecular chaperone function at low temperatures and proteolytic activity at elevated temperatures. To investigate the mechanism of functional switch to pretense, we have determined the crystal structure of the N-terminal protease domain (PD) of HtrA from Thermotoga maritima. HtrA PD shares the same fold with chymotrypsin-like serine professes. However, crystal structure suggests that HtrA PD is not an active pretense at current state since its active site is not formed properly and blocked by an additional helical lid. On the surface of the lid, HtrA PD has hydrophobic patches that could be potential substrate binding sites for molecular chaperone activity. Present structure suggests that the activation of the proteolytic function of HtrA PD at elevated temperatures might occur by the conformational change.

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Tissue and Immune Responses on Implanted Nanostructured Biomaterials

  • Khang, Dong-Woo;Kang, Sang-Soo;Nam, Tae-Hyun
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2009.05a
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    • pp.47.1-47.1
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    • 2009
  • Nanostructured biomaterials have increased those potential for utilizing in many medical applications. In this study, benefit of nanotechnology for the response with biological targets will be described in terms of size, effective surface area and surface energy (physical aspect). Also, correlations between physical and biological interactions (greater protein adsorption on nano surface roughness) will be discussed for understanding biocompatibility of nanostructured biomaterials including carbon nanotube composites and nanostructured titanium surfaces. In the application parts, various major tissue cells, such as bone, cartilage, vascular and bladder cell responses will be discussed with suggested nanomaterials. Lastly, immune responses with macrophage (adhesion and several major cytokines) on nanostructured biomaterials will be described for evasive immune response.

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Fibrin affects short-term in vitro human mesenchymal stromal cell responses to magneto-active fibre networks

  • Spear, Rose L.;Symeonidou, Antonia;Skepper, Jeremy N.;Brooks, Roger A.;Markaki, Athina E.
    • Biomaterials and Biomechanics in Bioengineering
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    • v.2 no.3
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    • pp.143-157
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    • 2015
  • Successful integration of cementless femoral stems using porous surfaces relies on effective periimplant bone healing to secure the bone-implant interface. The initial stages of the healing process involve protein adsorption, fibrin clot formation and cell osteoconduction onto the implant surface. Modelling this process in vitro, the current work considered the effect of fibrin deposition on the responses of human mesenchymal stromal cells cultured on ferritic fibre networks intended for magneto-mechanical actuation of in-growing bone tissue. The underlying hypothesis for the study was that fibrin deposition would support early stromal cell attachment and physiological functions within the optimal regions for strain transmission to the cells in the fibre networks. Highly porous fibre networks composed of 444 ferritic stainless steel were selected due to their ability to support human osteoblasts and mesenchymal stromal cells without inducing untoward inflammatory responses in vitro. Cell attachment, proliferation, metabolic activity, differentiation and penetration into the ferritic fibre networks were examined for one week. For all fibrin-containing samples, cells were observed on and between the metal fibres, supported by the deposited fibrin, while cells on fibrin-free fibre networks (control surface) attached only onto fibre surfaces and junctions. Initial cell attachment, measured by analysis of deoxyribonucleic acid, increased significantly with increasing fibrinogen concentration within the physiological range. Despite higher cell numbers on fibrin-containing samples, similar metabolic activities to control surfaces were observed, which significantly increased for all samples over the duration of the study. It is concluded that fibrin deposition can support the early attachment of viable mesenchymal stromal cells within the inter-fibre spaces of fibre networks intended for magneto-mechanical strain transduction to in-growing cells.

Global Transcriptional Analysis Reveals Upregulation of NF-${\kappa}B$-responsive and Interferon-stimulated Genes in Monocytes by Treponema lecithinolyticum Major Surface Protein

  • Lee, Sung-Hoon;Lee, Hae-Ri;Jun, Hye-Kyoung;Choi, Bong-Kyu
    • International Journal of Oral Biology
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    • v.36 no.2
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    • pp.91-101
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    • 2011
  • MspTL is the major surface protein of Treponema lecithinolyticum associated with periodontitis and endodontic infections. Our recent investigation revealed that MspTL induces proinflammatory cytokines and intercellular adhesion molecule 1 in THP-1 cells and periodontal ligament cells. In this study we conducted oligonucleotide microarray analysis to investigate the global transcriptional regulation in THP-1 cells stimulated with purified recombinant MspTL. MspTL upregulated the expression of 90 genes in THP-1 cells at least four fold, and the functions of these genes were categorized into adhesion, apoptosis/antiapoptosis, cell cycle/growth/differentiation, chemotaxis, cytoskeleton organization, immune response, molecular metabolism, proteolysis, signaling, and transcription. The majority of the modified genes are known to be NF-${\kappa}B$-responsive and interferon-stimulated genes (ISGs). The expression of 12 selected genes was confirmed by real-time RT-PCR. Because prostaglandin $E_2(PGE_2)$ is an important inflammatory mediator and Cox-2 was found to be induced by MspTL in the microarray analysis, we determined the level of $PGE_2$ in the culture supernatants of MspTL-treated cells and found that MspTL significantly increased $PGE_2$. Our results provide insight into the gene regulation of host cells in response to MspTL, and may contribute to the understanding of the molecular mechanism in periodontitis.