• Title/Summary/Keyword: RGD

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Surface Modification and Fibrovascular Ingrowth of Porous Polyethylene Anophthalmic Implants

  • Yang, Hee-Seok;Park, Kwi-Deok;Son, Jun-Sik;Kim, Jae-Jin;Han, Dong-Keun;Park, Byung-Woo;Baek, Se-Hyun
    • Macromolecular Research
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    • v.15 no.3
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    • pp.256-262
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    • 2007
  • The purpose of this study was to determine the effect of surface modification on the fibrovascular ingrowth into porous polyethylene (PE) spheres ($Medpor^{(R)}$), which are used as an anophthalmic socket implant material. To make the inert, hydrophobic PE surface hydrophilic, nonporous PE film and porous PE spheres were subjected to plasma treatment and in situ acrylic acid (AA) grafting followed by the immobilization of arginine-glycine-aspartic acid (RGD) peptide. The surface-modified PE was evaluated by performing surface analyses and tested for fibroblast adhesion and proliferation in vitro. In addition, the porous PE implants were inserted for up to 3 weeks in the abdominal area of rabbits and, after their retrieval, the level of fibrovascular ingrowth within the implants was assessed in vivo. As compared to the unmodified PE control, a significant increase in the hydrophilicity of both the AA-grafted (PE-g-PAA) and RGD-immobilized PE (PE-g-RGD) was observed by the measurement of the water contact angle. The cell adhesion at 72 h was most notable in the PE-g-RGD, followed by the PE-g-PAA and PE control. There was no significant difference between the two modified surfaces. When the cross-sectional area of tissue ingrowth in vivo was evaluated, the area of fibrovascularization was the largest with PE-g-RGD. The results of immunostaining of CD31, which is indicative of the degree of vascularization, showed that the RGD-immobilized surface could elicit more widespread fibrovascularization within the porous PE implants. This work demonstrates that the present surface modifications, viz. hydrophilic AA grafting and RGD peptide immobilization, can be very effective in inducing fibrovascular ingrowth into porous PE implants.

RGD Fixation of Film Surface and Synthesis of Copolymer Comprising Malic acid (Malic acid 함유 공중합체의 합성 및 필름표면의 RGD 고정화)

  • 이찬우;송경헌
    • Polymer(Korea)
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    • v.26 no.3
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    • pp.300-306
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    • 2002
  • To study the effect of polymer compositions on the cell adhesion, copolymers of 3-(S)-[(dodecyloxycarbonyl) methyl]-1,4-dioxane-2,5-dione (DMD) and L-lactide were made, where DMD was synthesized form L-malic acid (L-MA) and glycolic acid. Furthermore, the copolymerization of DMD and L-lactide was performed using tin(II) octanoate as a catalyst. As a result of fixing RGD on the copolymer films, the cell adhesive peptide was fixable on the surface of the film. It was found out that the amount of fixation of RGD also increases by the increase in the amount of MA unit introduction. Since it is gradually decomposed over a long period and neither remains nor accumulation occurs, glycolic acid-$\beta$-dodecylmalate -lactic acid (D-PGML) is greatly expected as a potential biomaterial with improved slow degradability.

NMR Structural Studies on Novel Disintegrin, Saxatilin from Gloydius saxatilis Venom

  • Shin, Joon;Lee, Dong-Hee;Hong, Sung-Yu;Chung, Kwang-Hoe;Kim, Doo-Sik;Lee, Weon-Tae
    • Journal of the Korean Magnetic Resonance Society
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    • v.11 no.1
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    • pp.10-23
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    • 2007
  • A new disintegrin protein named saxatilin was purified from Korean snake venom (Gloydius saxatilis). Saxatilin is a 73 residue small ploypeptide, which has a primary recognition motif in extracellular matrix, Arg-Gly-Asp (RGD) sequence. Data from inhibition activity assay for the ${\alpha}_v{\beta}_3$ integrin showed that saxatilin showed about 5000-fold higher activity than those of RGD peptides, suggesting that RGD sequence may not be sufficient to induce full cellular function of this site. The solution structures calculated from NMR data were well converged for backbone atoms except RGD loop. The structure revealed that most of tight turns are stabilized by medium range NOE contacts and the RGD motif is located far from the rigid core of the C-terminal domain. The three-dimensional fold and biological function of saxatilin are discussed with those of salmosin, which is a disintegrin protein derived from Agkistrodon halys brevicaudus.

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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.

Simple Purification of BA-RGD Protein Based on CaCl2/EDTA Treatment and Inclusion Body Washing (CaCl2/EDTA 및 비이온성 계면활성제 활용 Inclusion Body 정제법을 이용한 BA-RGD 단백질의 생산)

  • Song, Wooho;Byun, Chang Woo;Yoon, Minho;Eom, Ji Hoon;Choi, Yoo Seong
    • KSBB Journal
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    • v.30 no.6
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    • pp.291-295
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    • 2015
  • The limited productivity of natural shell matrix proteins has hampered the investigation of their biochemical properties and practical applications, although biominerals in nature obtained by organic-inorganic assemblies have attractive mechanical and biological properties. Here, we prepared a vector for the expression of a fusion protein of a shell matrix protein from Pinctada fucata (named as GRP_BA) with the GRGDSP residue. The fusion protein of BA-RGD was simply produced in E. coli and purified through sequential steps including the treatment with $CaCl_2$ and EDTA solution for cell membrane washing, mechanical cell disruption and the application of non-ionic surfactant of Triton X-100 for BA-RGD inclusion body washing. The production yield was approximately 60 mg/L, any other protein band was not observed in SDS-PAGE and it was estimated that above 97% endotoxin was removed compared to the endotoxin level of whole cell. This study showed this simple and easy purification approach could be applied to the purification of BA-RGD fusion protein. It is expected that the protein could be utilized for the preparation of biominerals in practical aspects.

Mechanism of Glucose Uptake on PMA Stimulated Neutrophils (PMA에 의한 중성구의 당섭취 기전 연구)

  • Paik, Jin-Young;Ko, Bong-Ho;Yoo, Man-Kil;Jin, Kwang-Ho
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.3
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    • pp.249-255
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    • 2007
  • While respiratory burst enhances neutrophil glucose utilization, many neutrophil functions are critically influenced by extracellular matrix interaction and phosphoinositide-3-OH kinase (PI3K) signaling. We thus evaluated the role of RGD integrin occupancy and PI3K inhibition on respiratory burst and [18F]FDG uptake of stimulated neutrophils. Human neutrophils were stimulated by 100 ng/mL phorbol-myristate-acetate (PMA), and respiratory burst was measured by cumulative luminescence with lucigenin. [18F]FDG uptake and total hexokinase activity was measured 20 min after PMA stimulation in the presence or absence of soluble RGD peptides (200 g/mL) and/or the PI3K inhibitor wortmannin (200 nM). PMA induced a 71.70.9 fold increase in neutrophil oxygen intermediate generation. [18F]FDG uptake was increased to $194.6{\pm} 3.7%$ and hexokinase activity to $145.0{\pm}2.0%$ of basal levels (both p<0.0005). RGD peptides attenuated respiratory burst activation to $35.6{\pm}0.2%$ (p<0.005), but did not inhibit stimulated [18F]FDG uptake or hexokinase activity. In contrast, without affecting respiratory burst activation, wortmannin inhibited PMA stimulated [18F]FDG uptake to $66.9{\pm}1.6%$ and hexokinase activity to $81.0{\pm}4.2%$ (both P<0.0005), demonstrating its dependence on PI3K activity. Neither RGD nor wortmannin reversed the other's inhibitory effect on stimulated [18F]FDG uptake and hexokinase activity or respiratory burst, which suggests the involvement of distinct signaling pathways. Neutrophil [18F]FDG uptake is enhanced by PMA through a mechanism that requires PI3K activity but is independent of integrin receptor occupancy or respiratory burst activation.

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Physical stability of arginine-glycine-aspartic acid peptide coated on anodized implants after installation

  • Huh, Jung-Bo;Lee, Jeong-Yeol;Jeon, Young-Chan;Shin, Sang-Wan;Ahn, Jin-Soo;Ryu, Jae-Jun
    • The Journal of Advanced Prosthodontics
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    • v.5 no.2
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    • pp.84-91
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    • 2013
  • PURPOSE. The aim of this study was to evaluate the stability of arginine-glycine-aspartic acid (RGD) peptide coatings on implants by measuring the amount of peptide remaining after installation. MATERIALS AND METHODS. Fluorescent isothiocyanate (FITC)-fixed RGD peptide was coated onto anodized titanium implants (width 4 mm, length 10 mm) using a physical adsorption method (P) or a chemical grafting method (C). Solid Rigid Polyurethane Foam (SRPF) was classified as either hard bone (H) or soft bone (S) according to its density. Two pieces of artificial bone were fixed in a customized jig, and coated implants were installed at the center of the boundary between two pieces of artificial bone. The test groups were classified as: P-H, P-S, C-H, or C-S. After each installation, implants were removed from the SRPF, and the residual amounts and rates of RGD peptide in implants were measured by fluorescence spectrometry. The Kruskal-Wallis test was used for the statistical analysis (${\alpha}$=0.05). RESULTS. Peptide-coating was identified by fluorescence microscopy and XPS. Total coating amount was higher for physical adsorption than chemical grafting. The residual rate of peptide was significantly larger in the P-S group than in the other three groups (P<.05). CONCLUSION. The result of this study suggests that coating doses depend on coating method. Residual amounts of RGD peptide were greater for the physical adsorption method than the chemical grafting method.

Effect of RGD peptide coating of implant titanium surface on human mesenchymal stem cell response (양극산화 티타늄 표면에 서로 다른 RGD 펩타이드 코팅 방법이 인간간엽줄기세포 반응에 미치는 영향)

  • Kim, Min-Su;Jeong, Chang-Mo;Jeon, Young-Chan;Ryu, Jae-Jun;Huh, Jung-Bo;Yun, Mi-Jung
    • The Journal of Korean Academy of Prosthodontics
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    • v.49 no.3
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    • pp.245-253
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    • 2011
  • Purpose: The aim of this in vitro study was to estimate surface characteristic after peptide coating and investigate biological response of human mesenchymal stem cell to anodized titanium discs coated with RGD peptide by physical adhesion and chemical fixation. Materials and methods: Fluorescence isothiocyanate (FITC) modified RGD-peptide was coated on the anodized titanium discs (diameter 12 mm, height 3 mm) using two methods. One was physical adhesion method and the other was chemical fixation method. Physical adhesion was performed by dip and dry procedure, chemical fixation was performed by covalent bond via silanization. In this study, human mesenchymal stem cell was used for experiments. The experiments consisted of surface characteristic evaluation after peptide coating, analysis about cell adhesion, proliferation, differentiation, and mineralization. Obtained data are statistically treated using Kruskal-Wallis test and Bonferroni test was performed as post hoc test (P=.05). Results: The evaluation of FE-SEM images revealed no diffenrence at micro-surfaces between each groups. Total coating dose was higher at physical adhesion experimental group than at chemical fixation experimental group. In cell adhesion and proliferation, RGD peptide coating did not show a statistical significance compared with control group (P>.05). In cell differentiation and mineralization, physical adhesion method displayed significantly increased levels compared with control group and chemical fixation method (P<.05). Conclusion: RGD peptide coating seems to enhance osseointegration by effects on the response of human mesenchymal stem cell. Especially physical adhesion method showed more effective than chemical fixation method on response of human mesenchymal stem cell.

Preliminary evaluation of new 68Ga-labeled cyclic RGD peptides by PET imaging

  • Shin, Un Chol;Jung, Ki-Hye;Lee, Ji Woong;Lee, Kyo Chul;Lee, Yong Jin;Park, Ji-Ae;Kim, Jung Young;Kang, Joo Hyun;An, Gwang Il;Ryu, Young Hoon;Choi, Jae Yong;Kim, Kyeong Min
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.2 no.2
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    • pp.118-122
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    • 2016
  • Integrin ${\alpha}_v{\beta}_3$ plays an important role in the tumor metastases and angiogenesis. Arginine-glycine-aspartate (RGD) peptide motif binds to the integrin ${\alpha}_v{\beta}_3$. General $^{68}Ga$-labeled cyclic RGD peptides was rapidly eliminated from the circulatory system by renal excretion because of its hydrophilic property. The purpose of this study was to develop a novel $^{68}Ga$-labeled cyclic RGD peptides, which could acquire enhanced PET tumor images with improved pharmacokinetics by adopting biphenyl group between chelator and RGD peptides. $^{68}Ga$-DOTA-2P-c(RGDyK) was demonstrated a 12% higher lipophilicity level than $^{68}Ga$-DOTA-c(RGDyK) as a reference compound. In the animal PET, $^{68}Ga$-DOTA-2P-c(RGDyK) represented relatively lower blood-clearance, and an increased signal to noise ratio compared to $^{68}Ga$-DOTA-c(RGDyK). From these perspective, $^{68}Ga$-DOTA-2P-c(RGDyK) could be a good candidate for in integrin ${\alpha}_v{\beta}_3$-expressed tumor imaging.