Hiroyoshi Kawakami

Aligned Electrospun Nanofiber Composite Membranes for Fuel Cell Electrolytes

We have synthesized the novel composite membranes composed of sulfonated polyimide nanofibers and sulfonated polyimide for proton exchange membrane fuel cell. It was clear that the polyimides within nanofiber were significantly oriented or aggregated when electrospun; as the result, the membrane stability, such as oxidative and hydrolytic stabilities, of the composite membrane was significantly improved with an increase in nanofiber, and oxygen permeability of the composite membrane also decreased when compared to that determined in the membrane without nanofibers. In addition, the proton conductivity of the membrane in the parallel direction indicated a significantly higher value when compared to that determined for the membrane in the perpendicular direction or for the membrane without nanofibers prepared with conventional solvent-casting method. Consequently, nanofibers proved to be promising materials as a proton exchange membrane and the composite membrane containing nanofibers may have potential application for use in fuel cells.

Gas transport properties in thermally cured aromatic polyimide membranes

Aromatic polyimide derived from 2,2′-bis(3,4-dicarboxyphenyl)hexafluoropropane dianhydride (6FDA) and 3,3′-diaminodiphenylsulfone (m-DDS) has been synthesized to facilitate the study of relationships between the polymer structure and the gas transport properties (permeability and selectivity). The gas permeability and selectivity of CO2, O2, N2, and CH4 for the 6FDA-m-DDS membranes cured at 150, 200, and 250°C have been determined at 35°C and at pressures up to 760 cmHg. The packing density and the fluorescence intensity of the 6FDA-m-DDS polyimide increased sharply with the increasing curing temperature. We propose that this behavior is associated with an increase in intermolecular and/or intramolecular interactions by a charge transfer complex formed in 6FDA-m-DDS containing an alternating sequence of electron donor and electron aceptor molecules. The effect of the microstructure of the thermally cured 6FDA-m-DDS membranes on their gas transport properties is discussed.

Surface modification of poly(L-lactic acid) affects initial cell attachment, cell morphology, and cell growth

The object of this study was to develop a highly porous scaffold to be used in regeneration of blood vessels, nerves, and other hollow tissues with small openings. Using the phase-inversion method and a mixture of water and methanol as a coagulating agent, we prepared highly porous flat membranes from poly(l-lactic acid) (PLLA) with numerous pores both on the surface and in the interior of the membranes. Chinese hamster ovary (CHO) cells were cultured on the membranes to evaluate initial cell adhesion, cell proliferation, and cell morphology. Adhesion of CHO cells to PLLA was poor: the cells adhered at approximately half the rate observed with a tissue culture polystyrene dish (TCPS). In contrast, adhesion of cells to PLLA treated with a low-temperature oxygen plasma was good; the adhesion rate was the same as that on TCPS. The rate of cell proliferation on the treated membranes was no different from that on the nontreated membranes, but cell morphologies were quite different. The cells on the nontreated membranes were small and round and proliferated separately from one another. In contrast, the cells on the plasma-treated membranes proliferated in close contact with other cells, spreading out extensively in sheet-like formations. Since the plasma treatment not only accelerated cell adhesion but also enabled cells to proliferate in the form of sheets resembling biological tissue, we believe that oxygen-plasma treatment is extremely effective for modifying surfaces of materials used for tissue regeneration.

Formation of surface skin layer of asymmetric polyimide membranes and their gas transport properties

Abstract The fabrication and gas-transport properties of asymmetric 6FDA-APPS membranes have been described in this study. The apparent skin-layer thickness strongly depended on the evaporation time, which was of the utmost importance for the formation of an asymmetric membrane with high gas permeance. Surface morphology of the membrane has been determined by atomic force microscopy (AFM). The surface roughness parameters of the membranes analyzed from AFM micrographs increased with a decrease in the apparent skin-layer thickness. Permeance and selectivity of CO 2 , O 2 , N 2 , and CH 4 for the asymmetric membranes at 35°C and at pressures up to 760 cmHg have been measured. The selectivities for (O 2 /N 2 ) and (CO 2 /CH 4 ) in the membrane with a skin layer of 34 nm were 6.1 at an O 2 permeance of 5.6 × 10 −5 cm 3 (STP)/cm 2 s cmHg and 39 at a CO 2 permeance of 2.7 × 10 −4 cm 3 (STP)/cm 2 s cmHg, respectively, without the necessity of an additional coating process. The asymmetric membrane exhibited gas selectivities equal to or even higher than those determined for the dense membrane.

Synthesis and Characterization of Alkylated Poly(1-vinylimidazole) to Control the Stability of its DNA Polyion Complexes for Gene Delivery

Poly(1-vinylimidazole) (PVIm) with alkylated imidazole groups has been synthesized as a pH-sensitive polycation to control the stability of its DNA polyion complexes for gene delivery. The resulting alkylated PVIm (PVIm-R) was water-soluble despite deprotonation of the imidazole groups at physiological pH, as determined by acid-base titration and solution turbidity measurements. Agarose gel retardation assay proved that the alkylated imidazole groups worked as anchor groups to retain DNA. Pyrene fluorescence measurement showed that the hydrophobic domain of the DNA complex with butylated PVIm (PVIm-Bu) increased after the protonation of imidazole groups of the PVIm-Bu to enhance the membrane disruptive activity. The PVIm-Bu exhibited no significant cytotoxicity in spite of the existence of cationic groups. The resulting PVIm-Bu/DNA complexes easily released DNA, as compared with the octylated PVIm, which was examined by competitive exchange with dextran sulfate. As a result, the PVIm-R/DNA complexes mediated efficient gene delivery, and the gene expression depended on the length and density of the alkyl chains. These results suggest that pH-sensitive PVIm-Rs control of the stability of DNA polyion complexes enhanced noncytotoxic gene delivery by the optimized alkylated imidazole groups.

Cell death by reactive oxygen species generated from water-soluble cationic metalloporphyrins as superoxide dismutase mimics

We investigated the effect on cell death of reactive oxygen species induced by water-soluble cationic metalloporphyrins with superoxide dismutase (SOD) activity. The SOD activity of 5,10,15,20-tetrakis(4-N-methylpyridyl)]porphine (MPy(4)P) containing Fe, Mn or Cu was measured using a cytochrome c assay by the xanthine/xanthine oxidase system and stopped-flow kinetic analysis. Cell viability of four cell lines treated with metalloporphyrins, mitomycin c (MMC), or cisplatin was estimated by a trypan blue exclusion assay. FeMPy(4)P with a high SOD activity showed a significant cytotoxicity compared with MMC and cisplatin, while CuMPy(4)P without SOD activity exhibited no cytotoxicity. However, MnMPy(4)P showing an SOD activity as high as that of FeMPy(4)P did not indicate cytotoxicity. These findings suggest that FeMPy(4)P as SOD mimic converts intracellular O2(*-) to H(2)O(2) and that it rapidly reacts with H(2)O(2) to form *OH, causing DNA damage and inducing cell death. On the other hand, MnMPy(4)P did not participate in the Fenton reaction, so that DNA damage in the cells treated with MnMPy(4)P was not observed. In addition, the cytotoxicity by the metalloporphyrin was inversely correlated with the SOD activity of the cells and the selective damage at cellular and DNA levels was confirmed. We believe that for an anticancer drug with antioxidant ability O(2)(*-) is useful as a target molecule to induce selective cell death between cancer and normal cells and that metalloporphyrins showing SOD activity and Fenton-like reaction are a new class of anticancer agents.

Effect of polyethylene glycol on preparation of rifampicin-loaded PLGA microspheres with membrane emulsification technique

Monodisperse poly(lactide-co-glycolide) (PLGA) microspheres containing rifampicin (RFP), anti-tubercle drug, as hydrophobic model drug were prepared by solvent evaporation method with a membrane emulsification technique using Shirasu Porous Glass (SPG) membranes. Five kinds of rifampicin-loaded PLGA (RFP/PLGA) microspheres with different sizes were prepared by changing pore size of the membranes. Effect of polyethylene glycol (PEG) added to polyvinyl alcohol (PVA) solution (continuous phase) upon the monodispersity of microspheres was studied. PEG was used as a stabilizer for microspheres dispersing in PVA solution. The most suitable molecular weight of PEG as a stabilizer was 20,000. RFP/PLGA microspheres prepared with PEG20000 were apparently more uniform than those prepared without PEG. The yield of RFP/PLGA microspheres was 100%. The initial burst observed in the release of RFP from RFP/PLGA microspheres was suppressed by the addition of PEG.

Selective cell death by water-soluble Fe-porphyrins with superoxide dismutase (SOD) activity

We investigated the effect on cell death of reactive oxygen species induced by [[5,10 (or 5,15)-bis(N-methyl-4-pyridyl)-15,20 (or 10,20) diphenyl]porphinato]iron (cis-FeMPy(2)P(2)P or trans-FeMPy(2)P(2)P) with SOD activity. The SOD activities of the cis-FeMPy(2)P(2)P and trans-FeMPy(2)P(2)P were measured using stopped-flow kinetic analysis. The cell viability of four cell lines treated with cis-Fe-porphyrin, trans-Fe-porphyrin, mitomycin c (MMC), or cisplatin was estimated by the alamar blue exclusion assay of the modified MTT method. The amount of cis-FeMPy(2)P(2)P and trans-FeMPy(2)P(2)P in the Walker 256 cultured for 24 h was 4.0 and 2.6 fmolcell(-1), respectively, indicating that the plasma membrane permeability of the Fe-porphyrins depended on their structure. Cis-FeMPy(2)P(2)P selectively killed Walker 256 and H-4-II-E as cancer cells but not FR and BRL-3A as normal cells and showed a significant cytotoxicity for the cancer cells compared with trans-FeMPy(2)P(2)P, MMC and cisplatin. We believe that cis-FeMPy(2)P(2)P as an SOD mimic converts intracellular O(2)(*-) to H(2)O(2) and that H(2)O(2) or *OH causes DNA damage and induces cell death. This result suggests that for the SOD mimic, O(2)(*-) may be useful as a target molecule to induce selective cell death between cancer and normal cells and that a metalloporphyrin having SOD activity is a new class of anticancer agents.

Phosphoric acid-doped sulfonated polyimide and polybenzimidazole blend membranes: high proton transport at wide temperatures under low humidity conditions due to new proton transport pathways

We now demonstrate a new class of a polymer proton exchange membrane for fuel cells, capable of combining low cost and high proton conductivity under low humidity conditions. The novel phosphoric acid (PA)-doped sulfonated polyimide (SPI) and polybenzimidazole (PBI) blend membrane enables proton conductivities of approximately 0.5 and 0.1 S cm−1 at 120 °C and 45%RH and at 30 °C and 30%RH, respectively. These conductivities were approximately two orders of magnitude higher than those of Nafion 117 due to a new proton transport pathway between phosphoric acid and sulfonic acid in the blend membrane. In addition, the membrane showed no signs of performance degradation at 120 °C and 0%RH after 1000 hours of operation, and maintained a proton conductivity value of more than 0.01 S cm−1. Consequently, this material shows promise as a novel proton exchange membrane and may have potential applications for use in fuel cells.

Carboxymethyl poly(L-histidine) as a new pH-sensitive polypeptide to enhance polyplex gene delivery.

Carboxymethyl poly( l-histidine) (CM-PLH) as a new pH-sensitive polypeptide has enhanced polyplex gene delivery. Agarose gel retardation assay and zeta potential measurement proved that the anionic CM-PLH at physiological pH coated the PEI/DNA binary complexes. The resulting CM-PLH/PEI/DNA ternary complexes showed the gene expression value 300 times higher than that of the PEI/DNA binary complexes. These results suggest that the synergistic effect of the pH-sensitive imidazole groups at endosomal pH and the anionic carboxymethyl groups at physiological pH in the CM-PLH enhanced polyplex gene delivery.