• 제목/요약/키워드: Extracellular matrices

검색결과 31건 처리시간 0.029초

Effects of Silkworm Hemolymph and Cartilage-specific Extracellular Matrices on Chondrocytes and Periosteum-derived Progenitor Cells

  • Shin, Hyun-Chong;Choi, Yong-Soo;Lim, Sang-Min;Lee, Chang-Woo;Kim, Dong-Il
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제11권4호
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    • pp.364-367
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    • 2006
  • In order to enhance the repair of defects in articular cartilage via cell therapy with autologous chondrocytes, as well as with periosteum-derived progenitor cells (PDPCs), silkworm hemolymph (SH) and a variety of cartilage-specific extracellular matrices (ECMs) including type II collagen, proline, chondroitin 4-sulfate, and chondroitin 6-sulfate were assessed with regard to their efficacy as media supplements. SH, a known anti-apoptotic agent, was found to enhance cell growth, as was shown by the results of a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay. According to the results of reverse transcriptase polymerase chain reaction (RT-PCR) analyses, the cartilage-specific ECMs were found to stimulate the expression of hyaline cartilage-specific genes, most notably type II collagen and Sox9, in monolayer cultures of PDPCs.

Purification of the Candida utilis Extracellular Invertase using Affinity Chromatography

  • Ginalska, G.;Belcarz, A.;Lobarzewski, J.;Leonowicz, A.;Cho, Nam-Seok
    • Journal of the Korean Wood Science and Technology
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    • 제30권3호
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    • pp.12-17
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    • 2002
  • The extracellular invertase (EC 3.2.1.26) (Candida utilis) preparation was obtained from the liquid medium after desalting and freeze drying. This prepared enzyme was used for the comparative purification on 4 activated matrices by liquid column affinity chromatography method. In this method there were used controlled porous glass (CPG) silanized covalently activated by keratin, silanized silica gel and silica gel covalently covered by keratin. It was found that the invertase purification process was better using both CPG matrices (silanized CPG and keratin activated CPG) than these with two silica gel supports. Also the elution coefficient of the invertase from the two CPG columns was about 93 to 94%. Two silica gel supports found to be superior in terms of purification efficiency. The invertase purification process was confirmed by PAGE electrophoresis.

Fibronectin-Dependent Cell Adhesion is Required for Shear-Dependent ERK Activation

  • Park, Heonyong;Shin, Jaeyoung;Lee, Jung Weon;Jo, Hanjoong
    • Animal cells and systems
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    • 제8권1호
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    • pp.27-32
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    • 2004
  • Endothellial cells are subjected to hemodynamic shear stress, the dragging force generated by blood flow. Shear stress regulates endothelial cell shape, structure, and function, including gene expression. Since endothelial cells must be anchored to their extracellular matrices(ECM) for their survival and growth, we hypothesized that ECMs are crucial for shear-dependent activation of extracellular signalactivated regulated kinase(ERK) that is important for cell proliferation. Shear stress-dependent activation of ERK was observed in cells plated on two different matrices, fibronectin and vitronectin(the two most physiologically relevant ECM in endothelial cells). We then treated bovine aortic endothelial cells(BAECs) with Arg-Gly-Asp(RGD) peptides that block the functional activation of integrin binding to fibronectin and vitronectin, and a nonfunctional peptide as a control. Treatment of cells with the RGD peptides, but not the control peptide, significantly inhibited ERK activity in a concentration-dependent manner. This supports the idea that integrin adhesion to the ligands, fibronectin and vitronectin, mediates shear stress-dependent activation of ERK. Subsequently, whereas antagonists of vitronectin(LM 609, an antibody for integrin ${\alpha}_{\gamma}$/${\beta}_3$ and XT 199, an antagonist specific for integrin ${\alpha}_{\gamma}$/${\beta}_3$) did not have any effect on shear-dependent activation of ERK, antagonists of fibronectin(a neutralizing antibody for integrin ${\alpha}_5$/${\beta}_1$or ${\alpha}_4$${\beta}_1$ and SM256) had an inhibitory effect. These results clearly demonstrate that mechanoactivation of ERK requires anchoring of endothelial cells to fibronectin through integrins.

Investigation of Cell-Matrix Interactions Using a FRET Technique

  • Shahbuddin, Munira B.;Park, Hong-Hyun;Lee, Jae-Won;Park, So-Yeon;Lee, Kuen-Yong
    • Bulletin of the Korean Chemical Society
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    • 제30권8호
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    • pp.1817-1820
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    • 2009
  • Controlling cell-matrix interactions is critical in regulating cell phenotypes for tissue engineering applications. Cellular adhesion to synthetic extracellular matrices (ECMs) can be enhanced by introduction of adhesion ligands to the matrices. We tested the hypothesis that biophysical cues such as ligand organization in synthetic ECMs play an important role in modulating cell responses to the microenvironment. To investigate and monitor cell-matrix interactions, we used a fluorescence resonance energy transfer (FRET) technique with cell-interactive polymers generated by conjugating a peptide with the sequence of arginine-glycine-aspartic acid (RGD) to alginate hydrogels.

Histological, Physical Studies after Xenograft of Porcine Ear Cartilage

  • Ryu, Yong Ah;Jin, Meiying;Kang, Nakheon
    • 대한두개안면성형외과학회지
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    • 제18권3호
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    • pp.155-161
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    • 2017
  • Background: Because of the relatively similar size of organs to human and the physiological and structural similarities, the use of porcine as xenograft donors is progressing very actively. In this study, we analyzed the characteristics of porcine ear cartilage and evaluated its suitability as graft material in reconstructive and cosmetic surgery. Methods: The auricular cartilage was harvested from two pigs, and subjected to histological examination by immunohistochemical staining. To determine the collagen content, samples were treated with collagenase and weight changes were measured. After sterilization by irradiation, the samples were grafted into rats and stained with Hematoxylin and Eosin and Masson Trichrome to observe inflammation and xenograft rejection. Results: In IHC staining, extracellular matrices were mainly stained with type II collagen (20.69%), keratin sulfate (10.20%), chondroitin sulfate (2.62%), and hyaluronic acid (0.84%). After collagenase treatment, the weight decreased by 68.3%, indicating that about 70% of the porcine ear cartilage was composed of collagen. Upon xenograft of the sterilized cartilages in rats, inflammatory cells were observed for up to 2 months. However, they gradually decreased, and inflammation and reject-response were rarely observed at 5 months. Conclusion: The porcine ear cartilage was covered with perichondrium and cellular constituents were found to be composed of chondrocytes and chondroblasts. In addition, the extracellular matrices were mainly composed of collagen. Upon xenograft of irradiated cartilage into rats, there was no specific inflammatory reaction around the transplanted cartilage. These findings suggest that porcine ear cartilage could be a useful alternative implant material for human cosmetic surgery.

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

  • 박병윤;서상우;이원재;류창우;나동균;손현주;박종철
    • Archives of Plastic Surgery
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    • 제32권2호
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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.

Bioactive Polyglycolic Acid (PGA) or Polylactic Acid (PLA) Polymers on Extracellular Matrix Mineralization in Osteoblast-like Mc3T3-E1 Cells

  • Cho, Young-Eun;Kim, Hye-Jin;Kim, Yong-Ha;Choi, Jae-Won;Kim, Youn-Jung;Kim, Gab-Joong;Kim, Jin-Su;Choi, Sik-Young;Kwun, In-Sook
    • Nutritional Sciences
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    • 제9권4호
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    • pp.233-239
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    • 2006
  • Porous matrices of bioactive polymers such as polyglycolic acid (PGA) or polylactic acid (PLA) can be used as scaffolds in bone tissue growth during bone repair process. These polymers are highly porous and serve as a template for the growth and organization of new bone tissues. We evaluated the effect of PGA and PLA polymers on osteoblastic MC3T3-E1 cell extracellular mineralization. MC3T3-E1 cells were cultured in a time-dependent manner -1, 15, 25d as appropriate - for the period of bone formation stages in one of the five culture circumstances, such as normal osteogenic differentiation medium, PGA-plated, fetal bovine serum (FBS)-plated, PGA/FBS-coplated, and PLA-plated For the evaluation of bone formation, minerals (Ca, Mg, Mn) and alkaline phosphatase activity, a marker for osteoblast differentiation, were measured Alizarin Red staining was used for the measurement of extracellular matrix Ca deposit During the culture period, PGA-plated one was reabsorbed into the medium more easily and faster than the PLA-plated one. At day 15, at the middle stage of bone formation, cellular Ca and Mg levels showed higher tendency in PGA- or PLA-plated treatments compared to non-plated control and at day 25, at the early late stage of bone formation, all three cellular Ca, Mg or Mn levels showed higher tendency as in order of PGA-related treatments and PLA-plated treatments, compared to control even without significance. Medium Ca, Mg or Mn levels didn't show any consistent tendency. Cellular ALP activity was higher in the PGA- or PLA-plated treatments compare to normal osteogenic medium treatment PGA-plated and PGA/FBS-plated treatments showed better Ca deposits than other treatments by measurement of Alizarin Red staining, although PLA-plated treatment also showed reasonable Ca deposit. The results of the present study suggest that biodegradable material, PGA and also with less extent for PLA, can be used as a biomaterial for better extracellular matrix mineralization in osteoblastic MC3T3-E1 cells.

Crosstalk between integrin and receptor tyrosine kinase signaling in breast carcinoma progression

  • Soung, Young-Hwa;Clifford, John L.;Chung, Jun
    • BMB Reports
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    • 제43권5호
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    • pp.311-318
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    • 2010
  • This review explored the mechanism of breast carcinoma progression by focusing on integrins and receptor tyrosine kinases (or growth factor receptors). While the primary role of integrins was previously thought to be solely as mediators of adhesive interactions between cells and extracellular matrices, it is now believed that integrins also regulate signaling pathways that control cancer cell growth, survival, and invasion. A large body of evidence suggests that the cooperation between integrin and receptor tyrosine kinase signaling regulates certain signaling functions that are important for cancer progression. Recent developments on the crosstalk between integrins and receptor tyrosine kinases, and its implication in mammary tumor progression, are discussed.