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Biomedical subjects

Kazunori Shimizu

Publications and source records attributed to Kazunori Shimizu.

7 recordsLinked to original sources
PubMed2007

Construction of multi-layered cardiomyocyte sheets using magnetite nanoparticles and magnetic force.

Heart tissue engineering requires construction of three-dimensional (3-D) tissues composed of cardiomyocytes (CMs) that are tightly connected to each other. The aim of this study was to construct "scaffold-less" multi-layered 3-D CM sheets using magnetic force-based tissue engineering (Mag-TE) and to evaluate the cell-to-cell functional connections within the CM sheets. Original magnetite cationic liposomes (MCLs) with a positive surface charge (which facilitate adsorption to the target cell surface) were taken up by CMs that were isolated from 2-day-old Wistar rats. When MCLs were added to the medium of CMs at magnetite concentrations of 25, 50, and 100 pg per cell, subsequent measurements showed that 7.2, 13.2, and 27.3 pg of magnetite were taken up per cell, respectively, after 4 h incubation at 37 degrees C. Further, no toxicity was observed after a 24 h incubation period. Using magnetically labeled CMs (magnetite concentration, 100 pg/cell), multi-layered CM sheets were constructed. Immunofluorescent staining of connexin43 demonstrated the presence of gap junctions within the CM sheets that were constructed by Mag-TE. Moreover, electrical connections within the CM sheets constructed by Mag-TE were confirmed using extracellular potential mapping. These results indicate that Mag-TE is a viable methodology for heart tissue engineering.

Animals↗
PubMed2006

Increase of organic solvent tolerance by overexpression of manXYZ in Escherichia coli.

Transcriptional analysis has been investigated to detect the genes involved in organic solvent tolerance. A time course of gene expression profiles of Escherichia coli after exposure to organic solvents revealed that the expression levels of manX, manY, and manZ genes were strongly upregulated by 13.2-, 10.0-, and 7.0-folds, respectively, after 2 min and then decreased after 10 min. Organic solvent tolerance of E. coli was investigated by inducing overexpression of manX, manY, and manZ genes and manXYZ operon that encode a sugar transporter of the phosphotransferase system. Although the expression of manX, manY, and manZ alone was not effective, the organic solvent tolerance level was increased by the expression of manXYZ. The intracellular hexane level was kept lower in E. coli cells overexpressing manXYZ after 4 h of incubation with hexane.

Bacterial Adhesion↗
PubMed2006

Enhanced cell-seeding into 3D porous scaffolds by use of magnetite nanoparticles.

To engineer functional tissues, a large number of cells must be successfully seeded into scaffolds. We previously proposed a methodology for tissue engineering using magnetite nanoparticles and magnetic force, which we termed "Mag-TE." In the present study, we applied the Mag-TE technique to a cell seeding process and have termed the technique "Mag-seeding." The cell-seeding efficiency of NIH/3T3 fibroblasts (FBs) by Mag-seeding was investigated using six types of commercially available scaffolds (5 collagen sponges and 1 D,D-L,L polylactic acid sponge) having various pore sizes. FBs were magnetically labeled with our original magnetite cationic liposomes (MCLs), which have a positive surface charge, to improve adsorption onto the cell surface. FBs labeled with MCLs were seeded onto a scaffold, and a magnet (4 kG) was placed under the scaffold. Mag-seeding facilitated successful cell seeding into the deep internal space of the scaffolds. Cell-seeding efficiency increased significantly in all scaffolds when compared to those without magnetic force. Moreover, when a high-intensity magnet (10 kG) was used, cell-seeding efficiency was significantly enhanced. These results suggest that Mag-seeding is a promising approach for tissue engineering.

Animals↗
PubMed2005

Magnetic force-based mesenchymal stem cell expansion using antibody-conjugated magnetoliposomes.

Recently, there has been an accumulation of evidence indicating that human mesenchymal stem cells (MSCs, multipotent cells resident in the bone marrow) are useful for autologous cell transplantation. However, only small numbers of MSCs have been obtained in bone marrow aspirates. We have developed a novel methodology for enriching and proliferating MSCs from bone marrow aspirates using antibody-conjugated magnetoliposomes (AMLs). The AMLs are liposomes conjugated to anti-CD105 antibody (immunoliposomes) and contain magnetite nanoparticles (diameter 10 nm). In the present study, the AMLs were added to a small volume (1 mL) of human bone marrow aspirate. After a 1-h incubation period, the bone marrow aspirates containing AMLs were seeded into 10-cm tissue culture dishes, and a disk-shaped magnet (diameter 2.2 cm; height 1 cm; 4000 Gauss) was positioned under the dish to enrich MSCs by magnetic force. The MSCs proliferated, forming colonies at the site where the magnet was positioned. In contrast, no colonies and very few viable cells were observed in ordinary culture based on plastic-adherent tendencies of cells without use of AMLs. These results suggest that this AML culture method can rapidly and efficiently expand a small number of MSCs into numbers suitable for clinical application.

Animals↗
PubMed2005

Discovery of glpC, an organic solvent tolerance-related gene in Escherichia coli, using gene expression profiles from DNA microarrays.

Gene expression profiles were collected from Escherichia coli strains (OST3410, TK33, and TK31) before and after exposure to organic solvents, and the six genes that showed higher gene expression were selected. Among these genes, glpC encoding the anaerobic glycerol-3-phosphate dehydrogenase subunit C remarkably increased the organic solvent tolerance.

Cyclohexanes↗
PubMed2005

[Aortic aneurysm].

Thoracic and abdominal aortic aneurysms have potential risk of sudden death, because they have a tendency of rupture. The risk of rupture is high, if they are saccular type or the diameters of them are large enough. Acute aortic dissection has a high risk of sudden death due to cardiac tamponade, if the dissection involves the ascending aorta.

Aortic Aneurysm↗