Papers by Author: Shigeaki Abe

Abstract: In this study, we investigated in cytocompativility and controlled release behavior of a model drug from nanoporous silica microparticles (NPSM). When Mouse osteoblastic cells (MC3T3-E1) were exposed to NPMSs, they indicated excellent cell viability. From NPSM contained in dental glass ionomer cement (NPMS-GIC), they can release cationic model drug molecules gradually in water for 2 weeks when they were contained in GIC. In contrast, GIC without NPSM specimens released the molecule only a couple of days. In the case of inionic molecules, their release were alos determined only a couple of days. These results suggest that NPSM can be used as a sustainable drug-release system in dental and medical fields

Abstract: We synthesized the inorganic/organic hybrid nanocrystal (EHA) by hydroxyapatite (HA) nanocrystal growth under the existence of tris(2,2,6,6-tetramethyl-3,5-heptanedionato)europium(III) (EuTH) complex. Then, folic acid derivative (FA-NHS: folate N-hydroxysuccinimidyl ester) as the targeting ligand for the HeLa cancer cells was immobilized on the EHA by the mediation of 3-aminopropyltriethoxysilane (APTES) and methyltriethoxysilane (MTES). When the FA-NHS molecular occupancy ratio on the EHA surface is around 3 to 5 %, the strong luminescence from the f-f transition of the Eu³⁺ ion and luminescence associated with energy transfer between the EuTH-FA monomer near 518 nm were observed. Moreover, the dispersibility in phosphate buffer saline was confirmed with immobilizing the positively-charged FA-NHS. The affinity and non-cytotoxicity between the nanocrystals and HeLa cancer cells were confirmed for 3 days. The red luminescence from the cells could be observed by fluorescence microscopy and the luminescence spectra.

Abstract: Controlled ion release property of nanoporous silica particles (NPS) were investigated using cationic fluorescent dye, rhodamine B. The dye was charged into a glass ionomer cements (GIC) pellet containing the particles and then the pellet were immersed into distilled water. The dye-release behavior was observed using a UV-vis. spectrophotometer. GIC containing NPS can release the dye for a couple of weeks, where as other samples released it only a few days. This result suggests that NPS has excellent sustained dye-release property.

Abstract: In this study, we investigated the cytocompatibility of ceramic nanoparticles on different types of cells. All ceramics nanoparticles investigated in this study except Copper oxide (CuO) exhibited good cytocompatibility and cell viability (90% or more) even at 20 ppm concentration. In contrast, CuO nanoparticles caused cell inflammation, and their effect depended on their particle size. Confocal fluorescence microscopy measurements indicated that some particles had penetrated into the cells. These results indicate that except CuO nanoparticles, all other ceramic nanoparticles reported herein exhibited excellent cytocompatibility even for lung epithelial cells.

Abstract: We modified the surface of organically modified montmorillonite (OMMT) with the carboxyl group using the silane coupling reaction and assessed its characteristics and cytocompatibility. Scanning electron microscope observations show that while the size and morphology of the obtained OMMT (OMMT-COOH) was unchanged, the surface of OMMT-COOH was coarser than that of OMMT. Fourier transform infrared spectra showed characteristic strong peaks at 1210 and 1630 cm⁻¹, corresponding to the peaks of the carboxyl group. X-ray diffraction analysis showed that the diffraction peak of OMMT-COOH corresponding to the (001) reflection was located at higher a 2θ value than that of OMMT. Results of the proliferation ratio and cell viability measurements indicated that the OMMT-COOH cytocompatibility is higher than that of OMMT. Based on these results, we conclude that cytocompatibility of montmorillonite would be improved by tuning the properties of the surface.

Abstract: Poly (L-lactic acid)/organically modified montmorillonite (PLLA/OMMT) nanocomposites were fabricated by a solution intercalation method. OMMT, modified with quaternary alkylammonium ion, was prepared by alkyltrialkoxysilane. The differential scanning calorimetry measurement revealed that the crystallization temperatures of PLLA/OMMT nanocomposites were at around 110 °C regardless of the existence of OMMT or the weight fraction of them. X-ray diffraction patterns suggested that the (001) diffraction was around 2θ = 2.5°. The TEM image showed variously expanded interlayer galleries of OMMT and partially exfoliated silicate layer unit in the matrix. Board-shaped specimens for mechanical property tests were fabricated by compression-molding at 190 °C (including 30 min annealing at 110 °C). The flexural modulus of the nanocomposites increased with increasing content of OMMT. Vickers hardness of the nanocomposites were almost same independent on weight fraction of OMMT.

Abstract: The reports on cytotoxic studies of carbon nanotubes (CNTs) increased exponentially. In the present study, we investigate murine macrophage RAW264.7 cell response for the CNTs immobilized on a polystyrene substrate. We prepared CNT-coated dishes, and estimate the interaction of RAW264.7 cells with CNTs by cell adhesion, proliferation assay, and measurement of TNF-α production. As a result, the highest cell adhesion and proliferation was observed on a commercially cell culture polystyrene dish, while CNT-coated dish indicate slightly lower activity of them. Moreover, amount of production of TNF-α on the CNT-coated dishes was considerable lower than that in the case of lipopolysaccharide (LPS) addition as a control. These results indicated that CNT-coated dishes could not show strong cytotoxicity for RAW264.7 cells in vitro.

Abstract: Cytotoxicity and cell behavior to micro / nanoparticles of TiO₂ and CuO was evaluated using the viability measurement and time-lapse observation. After cultured, osteoblastic cells MC3T3-E1 were exposed to particles. After 24 hour exposure, their morphology was observed using a SEM and the viability was measured. Cells exposed to TiO₂ indicated no or very low decrease of viability. The results were independent of the particle size. On the other hand, the viability of cells exposed to CuO decreased with the concentration, and showed the size dependence. The nanosized CuO indicated higher toxicity compared with micro-sized one. Dynamic behavior of cells exposed to nanoparticles, was succeeded to observe in a time-lapse method for 24 hours. The observation showed that the cells exposed to CuO became dead after forming a spherical shape. This is consistent with the image taken by SEM. Time-lapse observation made it possible to see the dynamic reaction process from cell contact to particles at first, the following cell activity response and finally to cell death, which revealed a considerably different morphology from the static cell observed after fixation by conventional method.

Abstract: We succeeded in determination the biodistribution of several nano-sized particles administered to mice through the tail vein. After administration, these particles were observed in the lung, liver and spleen. The distribution behaviors depend upon not only chemical species but also the particles size. To estimate their cytocompatibility, these particles were exposed to osteoblastic cell at several concentrations. When the concentration reached at 10 ppm, their viability remained at 80% or more even nano-sized particle contained rare earth element. Only CuO particles indicated the viability decrease. The effect depended on the particle size. These results suggested that the chemical species played a dominant key in the biodistribution and biocompatibility of nanoparticles compared with the size-effect.

Abstract: For application of carbon nanotubes (CNTs) as biomaterials, it is important to clarify the interaction between CNTs and proteins, which may affect on cell proliferation and differentiation. In this study, the adsorption behavior was investigated for representative proteins, bovine serum albumin, lysozyme and fetal bovine serum by using chromatography system. It was also aimed whether the different treatment conditions of CNTs affected the adsorption of proteins. CNTs used for this study had a straight shape and about 70nm in a diameter from SEM observation. There was not much difference between untreated and treated CNTs from SEM images. In chromatography experiments, all the proteins of albumin, lysozyme and serum were eluted immediately after injected to the column of untreated CNTs. Second elution appeared after buffer was changed from phosphate saline buffer to 25mM sodium hydroxide (NaOH). The same tendency was confirmed for CNTs with only acid treatment. However the eluted peaks became remarkably smaller after the column was changed to CNTs with thermal and twice acid treatments. These results indicated that treatment conditions for CNTs affected the adsorption behavior of proteins.