• Title/Summary/Keyword: cell attachment

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Surface Immobilization of $(1{\to}3)(1{\to}6)-{\beta}-glucan$ onto Biodegradable Polymer for Tissue Regeneration (조직 재생을 위한 Poly (D, L-lactide-co-glycolide) 표면에 $(1{\to}3)(1{\to}6)-{\beta}-glucan$ 고정에 대한 세포 점착 및 성장 효과)

  • Lee, S.G.;Lee, J.B.;Yu, S.M.;Park, J.C.;Choi, J.B.;Kim, J.K.
    • Journal of Biomedical Engineering Research
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    • v.27 no.5
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    • pp.218-223
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    • 2006
  • We examined the effects of ${\beta}$-glucan-reinforced PLGA film and scaffold on HDFs (human dermal fibroblast) attachment and proliferation. The PLGA films were prepared by simple solvent-casting method. The prepared films were grafted with $(1{\to}3)(1{\to}6)-{\beta}-glucan$ in various ratios after plasma treatment on surface. The surface of the film was characterized by contact angle measurement, scanning electron microscope (SEM), and Fourier-transform infrared spectrophotometer (FT-IR). The amount of $(1{\to}3)(1{\to}6)-{\beta}-glucan$ in the prepared film was indirectly determined by phenol-sulfuric acid method. The HDFs (Human dermal fibroblasts) were used to evaluate the cell attachment and proliferation on PLGA specimens before and after plasma/${\beta}-glucan$ treatment. The result showed that the plasma treated groups exhibited more mont of ${\beta}-glucan$ might be grafted than the non plasma treated groups. Cell attachment was significantly enhanced in the plasma/${\beta}-glucan$ grafted group after 4 hours incubation (p<0.05) due to the improved hydrophilicity and cytoactivity effect of the ${\beta}-glucan$. The cell proliferation of plasma/${\beta}-glucan$ (2mg/ml) grafted group was the highest rate among the groups (p<0.05).

Plasma Surface Modification of Patterned Polyurethane Acrylate (PUA) Film for Biomedical Applications

  • Yun, Young-Shik;Kang, Eun-Hye;Yun, In-Sik;Kim, Yong-Oock;Yeo, Jong-Souk
    • Proceedings of the Korean Vacuum Society Conference
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    • 2015.08a
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    • pp.223.2-223.2
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    • 2015
  • Polyurethane acrylate (PUA) has been introduced to utilize as a mold material for sub-100 nm lithography as it provides advantages of stiffness for nanostructure formation, short curing time, flexibility for large area replication and transparency for relevant biomedical applications. Due to the ability to fabricate nanostructures on PUA, there have been many efforts to mimic extracellular matrix (ECM) using PUA especially in a field of tissue engineering. It has been demonstrated that PUA is useful for investigating the nanoscale-topographical effects on cell behavior in vitro such as cell attachment, spreading on a substrate, proliferation, and stem cell fate with various types of nanostructures. In this study, we have conducted surface modification of PUA films with micro/nanostructures on their surfaces using plasma treatment. In general, it is widely known that the plasma treated surface increases cell attachment as well as adsorption of ECM materials such as fibronectin, collagen and gelatin. Effect of plasma treatment on PUA especially with surface of micro/nanostructures needs to be understood further for its biomedical applications. We have evaluated the modified PUA film as a culture platform using adipose derived stem cells. Then, the behavior of stem cells and the level of adsorbed protein have been analyzed.

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Fabrication of Cross-linked Nano-Fibrous Chitosan Membranes and Their Biocompatibility Evaluation

  • Nguyen, Thi-Hiep;Lee, Seong-Jin;Min, Young-Ki;Lee, Byong-Taek
    • Korean Journal of Materials Research
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    • v.21 no.2
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    • pp.125-132
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    • 2011
  • Fibrous chitosan membranes were fabricated as a substrate for skin applications using an electro-spinning process with different solvents and varying concentrations. Scanning electron microscopy (SEM) images confirmed that the formation of the chitosan fibrous membrane in trifluoroacetic acid was better than that in acetic acid. Fourier transform infrared spectroscopy showed that the chitosan fibers were cross-linked with glutaraldehyde, and that the cytotoxicity of the aldehyde groups was reduced by glycine and washing by NaOH and DI water. Chitosan cross-linked fibrous membranes were insoluble in water and could be washed thoroughly to wash away glycine and excess NaOH and prevent the infiltration of other water soluble bio-toxic agents using DI water. MTT assay method was employed to test the cytotoxicity of chitosan membranes during fabricating, treating and washing processes. After the dehydration of cell cultured chitosan membranes, cell attachment behavior on the material was evaluated using SEM method. Effect of the treatment processes on the biocompatibility of the chitosan membranes was shown by comparing of filopodium and lamellipodium of fibroblast cells on grown washed and unwashed chitosan fibrous membrane. The MTT assay and SEM morphology confirmed that the washed chitosan fibrous membrane increased cell attachment and cell growth, and decreased toxicity compared to results for the unwashed chitosan fibrous membrane.

A STUDY ON THE PHYSICAL PROPERTIES AND BIOLOGICAL CHARACTERISTICS OF DENTAL MAGNETIC ATTACHMENTS (치과용 자석부착장치의 물리적, 생물학적 특성에 관한 연구)

  • Lim, Yong-Sik;Kim, Yung-Soo;Kim, Chang-Whe;Kim, Yong-Ho
    • The Journal of Korean Academy of Prosthodontics
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    • v.37 no.1
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    • pp.1-22
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    • 1999
  • In order to investigate various physical and biological properties of dental magnetic attachments studies on retentive characteristics, corrosion properties, cytotoxicity of different magnetic systems for dental applications were done. For the study of retentive characteristics changes of retentive force by increasing air gap, wear properties of various attachment systems and loss of magnetic force by heat treatment were measured. Forte study of corrosion property of magnet covering metal electrochemical corrosion was done in artificial saliva and 0.9% NaCl solution between $-1,000mV_{SCE}\;and\;+1,000mV_{SCE}$. Anodic polarization curves are obtained about 6 types of samples and 2 types of solution. Corroded surfaces were examined with metallurgical microscope, scanning electron microscope and surface profilometer. For the study of cytotoxicity of magnetic attachment and its field cell growth and agar overlay test were done. The results of this study were as follows. 1. In Magnetic attachments using closed circuit retentive force at zero air gap was greater than magnets using open circuit, but decrease of retentive force by increasing air gap was also greater than open systems. 2. After 40,000 cycles of wear test all mechanical attachment resulted in varing degree of retention loss but in magnetic attachments no loss of retentive force was observed. 3. The magnetic attachment using Neodymium magnet showed early loss of retentive force about $200^{\circ}C$ but attachment using Samarium magnet showed some resistance to heat treatment and complete retention loss was observed about $500^{\circ}C$. The keeper was not influenced by heat treatment in retention. 4. In electrochemical corrosion test Dyna magnetic attachment covering metal showed the highiest corrosion resistance and Shiner magnet covering metal showed the least corrosion resistance and examination of corroded surface with metallurgical microscope, scanning electron micro-scope and surface profilometer also showed same results with anodic polarization corves. 5. The result of cell culture tests on the magnet covered with metal showed least recognizable cytotoxicity.

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Biochemical Analysis of Anagrapha falcifera NPV Attachment to Spodoptera frugiperda 21 Cells

  • PARK, JIN O;JAI MYUNG YANG;IN SIK CHUNG
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.361-364
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    • 1999
  • The binding characteristics of Anagrapha falcifera nuclear polyhedrosis virus (AtNPV) to Spodoptera frugiperda 21 (Sf21) cells were investigated. The cells displayed an affinity of 4.7×10/sup 10/M/sup -1/ with about 3,300 binding sites per cell. The biochemical nature of the AfNPV-binding sites on the cell surface was also partially identified. Our findings suggest that the binding-site moiety has a glycoprotein component, but that the direct involvement of oligosacccharides containing N-acetylglucosamine or sialic acid residues in binding is unlikely, and that AfNPV entry into Sf21 cells may be via receptor-mediated endocytosis.

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Effect of Citric Acid and Tetracycline HCl Root Conditioning on rhBMP-2 on Human Periodontal Ligament Fibroblast and Osteoblast cell (구연산과 테트라싸이클린으로 처리한 치근면에서 rhBMP-2가 치주인대섬유아세포와 골아세포의 활성에 미치는 영향)

  • Shim, Jung-Min;Han, Soo-Boo;Seol, Yang-Jo;Lee, Yong-Moo;Kim, Kyeong-Hwa;Kye, Seung-Beom;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.31 no.1
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    • pp.21-41
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    • 2001
  • The goal of Periodontal treatment is predictable periodontal regeneration. But until now, many products including GTR materials and growth factors are beyond of complete regeneration. BMP can induce ectopic bone formation when implanted into sites such as rat muscle and can greatly enhance healing of bony defects when applied exogenously. BMP can promote periodontal regeneration by their ability to stimulate new bone and new cementum formation. But little is known about optimal conditions required for the application. Root conditioning is used for bioacive root change so altered root surface provides a substrate that promotes chemotaxis, migration and attachment of peridontal cells encouraging connective attachment to the denuded root surface. The aim of this study is to investigate whether the acid conditioning change effect of rhBMP-2 on human periodontal ligament cell and osteoblast cell line. 288 periodontally involved root dentin slices are divided into 6 groups, each 48, 1)control, 2)treated with BMP, 3)treated with citric acid 4)treated with citric acid+BMP 5)treated with tetracycline 6)treated with TC+BMP. Each group was devided half, so 12 root dentin slices were seeded with periodontal ligament cells and 12 were seeded with osteoblasts. At day 2 and 7, cell number, protein assay, ALP activitiy was measured. To investigate morphology of cultured cells, SEM was employed. Statistical analysis was performed with SPSS 8.0 either t-test or ANOVA test. The results are ; Protein assay and cell number was slightly decreased in CA+BMP group compared to Ca group but it was not statistically significant and ALP activity was much more increased in CA+BMP group compared to CA group so there was no statistically significance between BMP and CA+BMP group and statistically significant compared to control group. Cell number and protein assay was slightly increased in TC group and ALP activity was much less the BMP group and CA group. Cell number and protein and ALP activity was not much increased in TC+BMP group. TC group and TC+BMP group showed cell morphology change in SEM. This results suggested that application of root surface with citric acid before BMP treatment might give better result in periodontal regeneration.

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Cloning of agrobacterium tumefaciens chromosomal virulence region (Agrobacterium tumefaciens의 염색체 DNA내에 존재하는 종양 유발 지역의 클로닝)

  • ;Cangelosi, G.A.;Nester, E.W.
    • Korean Journal of Microbiology
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    • v.28 no.2
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    • pp.104-108
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    • 1990
  • The chromosomal DNA of Agrobacterium tumefaciens contains the genes required for bacterial attachment to plant cell which is an essential atage in crown gall tumorigenesis by Ti-plasmid. In order to clone the genes, Agrobacterium tumefaciens strain A5512 was mutagenized by transposon Tn5 and two Agrobacterium tumefaciens mutants which are attachment-defective and nontumorigenic were isolated. From one of the two mutants, a chromosomal virulence region which was required for attachment to the plant cells was cloned.

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Immobilization of Arg-Gly-Asp (RGD) Sequence in Sugar-Containing Copolymer for Culturing Fibroblast Cells

  • Na, Kun;Park, Keun-Hong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.193-196
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    • 2004
  • The peptide Arg-Gly-Asp (RGD) was immobilized through their amino terminus to ends of a sugar bearing copolymer, producing a functional hybrid copolymer. Poly(N-p-vinylbenzyl-D-maltonamide-co-6-(p-vinylbenzamido)-hexanoic acid-g-GRGDS) [p(VMA-co-VBGRGDS)] promoted the attachment and growth of NIH fibroblast cells. The interaction between fibroblast cells and p(VMA-co- VBGRGDS) copolymer resulted in effective cell attachment, proliferation, and morphological changes by introduction of a GRGDS sequence. Moreover, when pretreated with soluble RGD monomer, attachment of fibroblast cells was suppressed approximately 50% from that of the p(VMA-co-VBGRGDS) surface.

Effects of $TGF-{\beta}1$ on Cellular Activity of Minocycline-Pretreated Human Periodontal Ligament Cells (($TGF-{\beta}$)이 Minocycline을 전처리한 사람 치주인대세포의 활성에 미치는 영향)

  • Yang, Seung-Oh;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.469-490
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    • 1996
  • The initial events required for periodontal regeneration is the attachment, spreading, and proliferation of appropriated cells at the healing sites. These have been reported that minocycline stimulates the attachment of periodontal ligament cells, and also $TGF-{\beta}1$ enhances the proliferation of periodontal ligament cells. The purpose of the present study was to evaluate the effects of $TGF-{\beta}1$ on the cellular activity of minocycline treated human periodontal ligament cells. Periodontal ligament cells were obtained from the explants of healthy periodontal ligaments of extracted 3rd molars or premolar teeth extracted from the patients for orthodontic treatment. The cells were cultured in minimal essential medium(${\alpha}-MEM$) supplemented with 10.000units/ml penicillin, $10,000{\mu}g/ml$ streptomycin and 10% FBS(fetal bovine serum) at $37^{\circ}C$ in a humidified atmosphere of 5% carbon dioxide and the 5th to the 8th passages of the cells were used. To evaluate the effect of minocycline on cell attachment, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After trypsinization, the cells were counted with hemocytometer and were taken photographs for observation of cellular morphology. To evaluate the effect of $TGF-{\beta}1$ on minocycline-pretreated periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4$ cells/ well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After incubation, 1 and 10ng/ml of $rh-TGF-{\beta}1$ were also added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay, DNA synthesis($^3H-thymidine\;assay$), and protein and collagen assay(3H-proline assay) were carried out. In the MIT assay, after 200ul MTT solutionlconeentration of 5mg/ml) were added to the each well of the 24-well plates and incubated for 3 hours, and 200 ul DMSO were added so as to dissolve insoluble blue formazan crystals which was formed in incubated period. Then it read plates on a ELISA reader. For mitogenic assay, 1 uCi/ml $^3H-thymidine$ was added to each well for the final 2 hours of the incubation periods. After labeling, the wells were washed 3 times with ice cold PBS and 4 times with 5% TCA to remove unincorporated label and precipitate the cellular DNA. DNA, with the incorporated $^3H-thymidine$, was solubilized with 500 ul of 0.1% NaOH/0.1% SDS. A 250 ul aliquot was removed from each well and placed in a scintillation vial with 4ml of scintillation cocktail. Using an liguid scintillation counter, counts per minute(CPM) were determined for each samples. 3 uCi/ml $^3H-proline$ was added to each well for the final 4 hours of the incubation periods and total protein and percent collagen synthesis were carried out. The results indicate that minocycline treated group with $100{\mu}g/ml$ concentration for 1.5 hours significantly increased than that of control in cell attachment, and cell process is also evident compared with that of control in cell morphology, and the cellular activity and DNA synthesis rate of cells treated minocycline and $TGF-{\beta}1$ significantly increased than that of control values, but were below to values of the $TGF-{\beta}1$ only treated group in MIT assay and $^3H-thymidine\;assay$, and the total protein synthesis of minocycline and $TGF-{\beta}1$ treated group also significantly increased than that of control values, but the percent collagen synthesis of tested group significantly decreased to compared with control. On the above the findings, the tested group of minocycline and $TGF-{\beta}1$ did not increase the effect on the cell activity than $TGF-{\beta}1$ only tested group and the tested group of minocycline inhibited cell activity. This results indicate that minocycline was effective on cell attachment in early stage, but it is harmful to cell activity, that inhibitory effect of minocycline was compensated with stimulatory effect of $TGF-{\beta}1$.

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