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Kei Kanie,
Yuji Narita,
Yingzi Zhao,
Fumiaki Kuwabara,
Makoto Satake,
Susumu Honda,
Hiroaki Kaneko,
Tomohiko Yoshioka,
Mina Okochi,
Hiroyuki Honda,
Ryuji Kato
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ABSTRACT: Controlling the balance of endothelial cells (ECs) and smooth muscle cells (SMCs) in blood vessels is critically important to minimize the risk associated with vascular implants. Extracellular matrix (ECM) plays a key role in controlling the cellular balance, suggesting a promising source of cell-selective peptides. To obtain EC- or SMC-selective peptides, we start by highlighting sequence differences found among ECM molecules as enriched targets for cell-selective peptides. We explored the EC- or SMC-selective performance of tripeptides that are specifically enriched only in collagen type IV, but not in types I, II, III, and V. Collagen type IV was chosen since it is the major ECM in the basement membrane of blood vessels, which separates ECs and SMCs. Among 114 collagen type IV-derived tripeptides pre-screened from in silico analysis, 22 peptides (19%) were found to promote cell-selective adhesion, as determined by peptide array. One of the best performing EC-selective peptides (Cys-Ala-Gly (CAG)) was mixed into an electrospun fine-fiber, a vascular graft material, for practical application. Compared to unmodified fiber, the CAG containing fiber surface was found to enhance adhesion of ECs (+190%) while limiting SMCs (-20%). These results are not only consistent with the hypothesis of ECM as a source of cell selective peptides, but also suggest a new genre of EC- or SMC-selective peptides for tissue engineering applications. Collectively, these findings favorably support the screening approach used to discover new peptides for these purposes.
Biotechnology and Bioengineering 02/2012; 109(7):1808-16. · 3.95 Impact Factor
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ABSTRACT: Both rapid endothelialization and the prevention of intimal hyperplasia are essential to improve the patency of small-caliber vascular grafts (SCVGs). Using the peptide array based screening system, we identified the peptide CAG (cysteine-alanine-glycine), which has a high affinity for endothelial cells and a low adhesive property for smooth muscle cells (SMCs). In this article, we report an in vivo analysis of novel vascular grafts that were constructed with a biodegradable polymer (poly-ε-caprolactone [PCL]) containing CAG peptide.
The novel SCVG, which measured 0.7 mm in diameter and 7 mm in length, was fabricated using the electrospinning technique. Carotid arterial replacement was performed on Sprague-Dawley rats using SCVGs with (group CAG) or without CAG (group C). Histologic and biochemical assessments were performed at 1, 2, and 6 weeks after implantation.
The ratio of endothelialization was significantly higher in group CAG compared with group C (CAG versus C, 64.4±20.0% versus 42.1±8.9% at 1 week; p=0.017; 98.2±2.3% versus 72.7±12.9% at 2 weeks; p=0.001; and 97.4±4.6% versus 76.7±5.4% at 6 weeks; p<0.001). Additionally, Western blot analysis showed that the level of endothelial nitric oxide synthase (eNOS) at 1 week in group CAG was significantly higher than that in group C (CAG versus C, 1.20±0.37 versus 0.34±0.16; p=0.013), and that α-smooth muscle actin (ASMA) at 6 weeks in group CAG was significantly lower than that in group C (CAG versus C, 0.89±0.06 versus 1.25±0.22; p=0.04).
The graft with CAG promoted rapid endothelialization and the potential for inhibition of intimal hyperplasia.
The Annals of thoracic surgery 11/2011; 93(1):156-63; discussion 163. · 3.74 Impact Factor
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ABSTRACT: Peptide arrays represent a powerful research tool, especially for the development of medical applications to screen peptides for the replacement of animal-derived proteins. This minireview focuses on the applicability of cellulose-bound peptide arrays to assay peptide- cell interactions. Moreover, the successful combination of peptide array and bioinformatics to overcome the limitation of library size in array-type screenings is reviewed.
Mini-Reviews in Organic Chemistry 04/2011; 8(2):171-177. · 2.41 Impact Factor
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ABSTRACT: Effective surface modification with biocompatible molecules is known to be effective in reducing the life-threatening risks related to artificial cardiovascular implants. In recent strategies in regenerative medicine, the enhancement and support of natural repair systems at the site of injury by designed biocompatible molecules have succeeded in rapid and effective injury repair. Therefore, such a strategy could also be effective for rapid endothelialization of cardiovascular implants to lower the risk of thrombosis and stenosis. To achieve this enhancement of the natural repair system, a biomimetic molecule that mimics proper cellular organization at the implant location is required. In spite of the fact that many reported peptides have cell-attracting properties on material surfaces, there have been few peptides that could control cell-specific adhesion. For the advanced cardiovascular implants, peptides that can mimic the natural mechanism that controls cell-specific organization have been strongly anticipated. To obtain such peptides, we hypothesized the cellular bias toward certain varieties of amino acids and examined the cell preference (in terms of adhesion, proliferation, and protein attraction) of varieties and of repeat length on SPOT peptide arrays. To investigate the role of specific peptides in controlling the organization of various cardiovascular-related cells, we compared endothelial cells (ECs), smooth muscle cells (SMCs), and fibroblasts (FBs). A clear, cell-specific preference was found for amino acids (longer than 5-mer) using three types of cells, and the combinational effect of the physicochemical properties of the residues was analyzed to interpret the mechanism.
Journal of Peptide Science 02/2011; 17(6):479-86. · 1.80 Impact Factor
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ABSTRACT: Angiotensin II (ang II), an octapeptide (DRVYVHPF), can regulate blood pressure by binding specifically to its receptor, AT1. A peptide (VVIVIY) in the first transmembrane of AT1 has been found, via peptide array technology, to have an affinity for ang II. In this study, the peptide P2, which contained the VVIVIY sequence, was mutated and screened using microbead display technology that utilized emulsion PCR and cell-free protein synthesis. After one round of screening, the binding activities of collected mutants were estimated using flow cytometry and a peptide array. Two of these exhibited improved association rate constants to ang II, compared to the P2 peptide.
Journal of Bioscience and Bioengineering 04/2010; 109(4):411-7. · 1.79 Impact Factor
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ABSTRACT: In a previous study, we reported the effectiveness of a bacterial strain showing monolayer adsorption to oil surfaces on microbial conversion at oil-water interfaces. In the present study, we screened wild type strains from our toluene-degrading bacterial library that showed similar properties and succeeded in obtaining five wild type strains that adsorb to oil surfaces as a cell monolayer. We investigated the effects of cultivation conditions on cell surface hydrophobicity of these five strains. The effects of substrate hydrophobicity and the porous carrier were not significant. By contrast, growth temperature greatly affected the cell surface hydrophobicity of all five strains, especially strain TIS1-127, which was phylogenetically identified as Pseudomonas sp. which is closely related to P. mosselii, P. monteilii, and P. plecoglossicida. Pseudomonas sp. TIS1-127 cells grown at 37 degrees C were determined by the kinetic microbial-adhesion-to-hydrocarbon (MATH) test to be fully hydrophilic (lower than 10% of MATH value) while the cells grown at 28 degrees C were highly hydrophobic (over 90% of MATH value). We investigated the effects of growth temperature on toluene conversion by TIS1-127 resting cells in single-phase batch cultivation and in two-liquid-phase partitioning reactors containing an emulsion consisting of 20% silicone oil and 80% cell suspension. In both cases, the cells grown at 28 degrees C showed much higher conversion ability than those grown at 37 degrees C. Toluene conversion followed Michaelis-Menten kinetics and the K(m) values for the cells grown at 28 degrees C were lower than 1/10 those for the cells grown at 37 degrees C.
Journal of Bioscience and Bioengineering 04/2009; 107(3):250-5. · 1.79 Impact Factor
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ABSTRACT: Prefoldin is a jellyfish-shaped hexameric chaperone that captures a protein-folding intermediate and transfers it to the group II chaperonin for correct folding. In this work, we characterized the organic solvent tolerance of Escherichia coli cells that overexpress prefoldin and group II chaperonin from a hyperthermophilic archeaum, Pyrococcus horikoshii OT3. The colony-forming efficiency of E. coli cells overexpressing prefoldin increased by 1,000-fold and decreased the accumulation of intracellular organic solvent. The effect was impaired by deletions of the region responsible for the chaperone function of prefoldin. Therefore, we concluded that prefoldin endows E. coli cells by preventing accumulation of intracellular organic solvent through its molecular chaperone activity.
Applied Microbiology and Biotechnology 07/2008; 79(3):443-9. · 3.42 Impact Factor
