PubMed Health⌕ Search

Biomedical subjects

Mitsuru Akashi

Publications and source records attributed to Mitsuru Akashi.

At least 19 recordsLinked to original sources
PubMed2007

Hydroxyapatite formed on/in agarose gel induces activation of blood coagulation and platelets aggregation.

We reported earlier that hydroxyapatite (HA) formed on/in agarose gels (HA/agarose) produced by alternate soaking process is a bone-filling material possessing osteoconductive and hemostatic effects. This process could allow us to make bone-like apatite that was formed on/in organic polymer hydrogel matrices. Here, we investigated the mechanism of hemostasis induced by HA/agarose and found that HA/agarose, but not agarose or HA powder, significantly shortened activated partial thromboplastin time (APTT). While HA/agarose did not show significant platelet aggregation, it markedly enhanced adenosine diphosphate (ADP)-induced platelet aggregation. Moreover, Western blot analysis revealed selective adsorption of vitronectin onto HA/agarose. We also observed marked differences between HA powder and HA/agarose in their XRD patterns. The crystallinity of HA powder was much higher compared to that of HA/agarose. Furthermore, 50-100 nm of tube-form aggregations was observed in HA powder on the other hand 100-200 nm of particles was observed in HA/agarose by SEM observation. Thus 100-200 nm of low crystallized particles on the surface structure of HA/agarose may play an important role in hemostasis. Our results demonstrated a crucial role of HA/agarose in the mechanism of hemostasis and suggested a potential role for HA/agarose as a bone-grafting material.

Adenosine Diphosphate↗
PubMed2006

Environmentally degradable, high-performance thermoplastics from phenolic phytomonomers.

Aliphatic polyesters, such as poly(lactic acid), which degrade by hydrolysis, from naturally occurring molecules form the main components of biodegradable plastics. However, these polyesters have become substitutes for only a small percentage of the currently used plastic materials because of their poor thermal and mechanical properties. Polymers that degrade into natural molecules and have a performance closer to that of engineering plastics would be highly desirable. Although the use of a high-strength filler such as a bacterial cellulose or modified lignin greatly increases the plastic properties, it is the matrix polymer that determines the intrinsic properties of the composite. The introduction of an aromatic component into the thermoplastic polymer backbone is an efficient method to intrinsically improve the material performance. Here, we report the preparation of environmentally degradable, liquid crystalline, wholly aromatic polyesters. The polyesters were derived from polymerizable plant-derived chemicals--in other words, 'phytomonomers' that are widely present as lignin biosynthetic precursors. The mechanical performance of these materials surpasses that of current biodegradable plastics, with a mechanical strength, sigma, of 63 MPa, a Young's modulus, E, of 16 GPa, and a maximum softening temperature of 169 degrees C. On light irradiation, their mechanical properties improved further and the rate of hydrolysis accelerated.

Journal Article↗
PubMed2006

Rapid and precise release from nano-tracted poly(N-isopropylacrylamide) hydrogels containing linear poly(acrylic acid).

We investigated the rapid and precise molecular release from hydrogels in response to dual stimuli. To achieve precise on/off drug release using thermoresponsive poly(N-isopropylacrylamide) hydrogels, we prepared nano-structured semi-IPNs, which consisted of thermosensitive PNIPAAm networks penetrated by pH-responsive poly(acrylic acid) (PAAc) linear chains and perforated to create nano-tracts as a molecular pathway. The present nano-tracted semi-IPNs show a rapid deswelling response to both temperature and pH. Model drug releases were investigated when simultaneous changes in temperature and pH were applied. We observed that the cationic drug was rapidly released and then abruptly discontinued from the nano-tracted semi-IPNs in response to the dual stimuli, and clear release and stopping cycles were repeatedly observed on successive steps. Moreover, the release rates and amount of drug released were controllable by the deswelling speed of the gels and the PAAc content inside the gels. This novel release system using the nano-tracted semi-IPNs may be useful for the high performance, pulsed release of molecules.

Acrylamides↗
PubMed2006

Effects of thermoresponsive coacervation on the hydrolytic degradation of amphipathic poly(gamma-glutamate)s.

Hydrolytic properties of thermoresponsive biopolymers with amphiphilic structures, gamma-PGA-P, were investigated. Hydrolysis was monitored in terms of molecular weight changes using GPC and spectroscopic measurements. The hydrolytic degradation of gamma-PGA-P was controlled by a change in the degree of propyl group conversion, reaction temperature, and/or reaction pH. The degradation was classified as the rapid elimination of propyl side chains and the moderate cleavage of peptide linkages in the backbone. Furthermore, hydrophobic environments established by the thermoresponsive coacervation of gamma-PGA-P60 solutions inhibited hydrolytic degradation reactions. Inversely, hydrolytic degradations increased coacervation temperatures. Kinetic studies of hydrolytic reactions suggest that the degradation rate of gamma-PGA-P60 solutions can be controlled by their thermoresponsivity. The hydrolysis reported here represents the first degradation rate controlled by thermoresponsive coacervation.

Hydrogen-Ion Concentration↗
PubMed2006

One-pot synthesis in aqueous system for water-soluble chitosan-graft-poly(ethylene glycol) methyl ether.

Chitosan is functionalized with poly(ethylene glycol) methyl ether (mPEG) at the amino and hydroxyl groups via a single step reaction in a homogeneous aqueous system. A chitosan aqueous solution obtained from the mixture of chitosan and hydroxybenzotriazole (HOBt) in water is a key factor in providing mild conditions to conjugate mPEG by using a carbodiimide conjugating agent. The reaction at ambient temperature for 24 h gives chitosan-g-mPEG with water solubility with mPEG content as high as 42%. This work demonstrates that a water-soluble chitosan-HOBt complex is an effective system for the preparation of chitosan derivatives via the aqueous system without the use of acids or organic solvents.

Biocompatible Materials↗
PubMed2006

One-step advanced preparation of surface-functional peptide nanospheres by the polymerization of L-phenylalanine N-carboxyanhydride with dual initiators.

Surface-grafted peptide nanospheres consisting of hydrophobic poly(L-phenylalanine) with hydrophilic poly(ethylene glycol) (PEG) grafts were successfully prepared by the one-step polymerization of L-phenylalanine N-carboxyanhydride with the dual initiators of hydrophobic n-butylamine and hydrophilic NH2-monoterminated PEG (NH2-PEG). The monodispersed peptide nanospheres were stably self-assembled during polymerization in a mixture of water/dimethyl sulfoxide to create a colloidal solution, but only aggregated in water or organic solvents. When n-butylamine or NH2-PEG was used as a solitary initiator, the peptide nanospheres were not formed. The peptide nanospheres showed high dispersion-stability in water, and their diameter was approximately 300 nm. Furthermore, the peptide nanospheres were well-redispersed in water, retaining the same diameter and monodispersity even after lyophilization. Peptide nanospheres with the functional carboxylic acid on their graft layer were also successfully prepared by the one-step preparation method. This one-step preparation method of surface-grafted peptide nanospheres will be useful as an advanced technology to develop biodegradable functional nanospheres.

Butylamines↗
PubMed2006

Hydrolytic and enzymatic degradation of nanoparticles based on amphiphilic poly(gamma-glutamic acid)-graft-L-phenylalanine copolymers.

Amphiphilic graft copolymers consisting of poly(gamma-glutamic acid) (gamma-PGA) as the hydrophilic backbone and L-phenylalanine ethylester (L-PAE) as the hydrophobic side chain were synthesized by grafting L-PAE to gamma-PGA. The nanoparticles were prepared by a precipitation method, and about 200 nm-sized nanoparticles were obtained due to their amphiphilic properties. The hydrolytic and enzymatic degradation of these gamma-PGA nanoparticles was studied by gel permeation chromatography (GPC), scanning electron microscopy (SEM), dynamic light scattering (DLS) and (1)H NMR measurements. The hydrolysis ratio of gamma-PGA and these hydrophobic derivatives was found to decrease upon increasing the hydrophobicity of the gamma-PGA derivates. The pH had an effect on the hydrolytic degradation of the polymer. The hydrolysis of the polymer could be accelerated by alkaline conditions. The degradation of the gamma-PGA backbone by gamma-glutamyl transpeptidase (gamma-GTP) resulted in a dramatic change in nanoparticle morphology. With increasing time, the gamma-PGA nanoparticles began to decrease in size and finally disappeared completely. Moreover, the gamma-PGA nanoparticles were degraded by four different enzymes (Pronase E, protease, cathepsin B and lipase) with different degradation patterns. The enzymatic degradation of the nanoparticles occurred via the hydrolysis of gamma-PGA as the main chain and L-PAE as the side chain. In the case of the enzymatic degradation of gamma-PGA nanoparticles with Pronase E, the size of the nanoparticles increased during the initial degradation stage and decreased gradually when the degradation time was extended. Nanoparticles composed of biodegradable amphiphilic gamma-PGA with reactive function groups can undergo further modification and are expected to have a variety of potential pharmaceutical and biomedical applications, such as drug and vaccine carriers.

Guanosine Triphosphate↗
PubMed2006

Enzyme-responsive release of encapsulated proteins from biodegradable hollow capsules.

Biodegradable hollow capsules encapsulating proteins were prepared via layer-by-layer assembly of chitosan and dextran sulfate on protein-entrapping mesoporous silica particles and the subsequent removal of the silica. The enzymatic degradation of the capsules in the presence of chitosanase was explored by scanning electron microscopy (SEM). With increasing time, the chitosan component was degraded by chitosanase, and the capsules began to deform and were finally destroyed. Sustained release of the encapsulated proteins was attained by using the enzymatic degradation of the hollow capsules. The release behavior was successfully manipulated by altering the charge of capsule surface.

Animals↗
PubMed2006

Novel biomineralization for hydrogels: electrophoresis approach accelerates hydroxyapatite formation in hydrogels.

As a first step toward hydroxyapatite (HAp) formation in agarose hydrogels, we have tailored the internal chemistry using an electrophoresis approach. HAp was formed using aqueous solutions of calcium chloride and disodium hydrophosphate, which were set in a conventional agarose electrophoresis apparatus. Calcium and phosphate ions provided cations and anions, respectively, and were shown to migrate into the agarose hydrogel toward the corresponding electrode side. HAp was formed after colliding with each ion. The time needed to reach complete HAp formation was 30 min, and 130 ng of HAp was formed in 1 mg of agarose hydrogel when the equilibrium swelling state was reached. The electrophoresis approach accelerated the HAp formation, and the linear velocity of 1 mm/min was shown to be roughly 15 times larger than that of simple diffusion (0.06 mm/min).

Crystallography, X-Ray↗
PubMed2006

Multifunctional conjugation of proteins on/into bio-nanoparticles prepared by amphiphilic poly(gamma-glutamic acid).

The present study focuses on nanoparticles composed of amphiphilic poly(gamma-glutamic acid) (gamma-PGA) as potential protein carriers. Amphiphilic graft co-polymers composed of y-PGA as the hydrophilic backbone and L-phenylalanine ethylester (L-PAE) as the hydrophobic segment were synthesized by grafting L-PAE to y-PGA using water-soluble carbodiimide (WSC). Due to their amphiphilic properties, the gamma-PGA-graft-L-PAE co-polymer formed monodispersed nanoparticles in water. The particle size of the nanoparticles composed of gamma-PGA-graft-L-PAE (gamma-PGA nanoparticles) was about 200 nm and showed a highly negative zeta potential. To evaluate their potential applications as multifunctional protein carrier, we prepared protein-entrapped gamma-PGA nanoparticles by encapsulation, covalent immobilization or physical adsorption methods. For this purpose, 11 different proteins with various molecular weights and isoelectric points (pI values) were used as model proteins. The encapsulation of the protein into the nanoparticles was observed for all tested proteins. The amount of protein covalently immobilized or adsorbed onto the nanoparticles showed different tends based on the molecular weight and pI of each protein. Positively charged proteins could be adsorbed onto the negatively charged nanoparticles by electrostatic interaction. Moreover, it was found that enzyme-encapsulated nanoparticles showed higher enzymatic activity than surface-immobilized nanoparticles. These results indicated that the enzymatic activity of the enzyme-entrapped nanoparticles was significantly affected by the conjugation method, and that encapsulation was the optimal method for the conjugation of proteins and nanoparticles. It is expected that the y-PGA nanoparticle will have great potential as multifunctional carriers in pharmaceutical and biomedical applications, such as drug and vaccine delivery systems.

Biocompatible Materials↗
PubMed2006

Preparation and characterization of stimuli-resistible hydrogels nano-coated with polyelectrolyte multilayer films.

Alginate (ALG) hydrogels were nano-coated with polyelectrolyte multilayer (PEM) films composed of poly(diallyldimethyl ammonium chloride) (PDDA) and poly(sodium 4-styrenesulfonate) (PSS). Non- and nano-coated ALG hydrogels were immersed into salt solutions (pH 6, ionic strength: 0.01 M), and the swelling behavior of these ALG hydrogels was observed by optical microscopy. The increases in the diameter of the 10-step coated ALG hydrogels were one fourth or less than that of the non-coated hydrogels. These results indicate that the characteristics of nano-coated ALG hydrogels remain unchanged independent of how the external environment is changed (e.g., pH, ionic strength). Consequently, tolerant ionic hydrogels may be prepared by the nano-coating of hydrogel surfaces with PEM films and may be useful for technological and biomedical applications.

Absorption↗
PubMed2006

Non-methylated CpG motif packaged into fusogenic liposomes enhance antigen-specific immunity in mice.

DNA rich in non-methylated CG motifs (CpGs) enhances induction of immune responses against co-administered antigen encoding genes. CpGs are therefore among the promising adjuvants known to date. However, naked plasmid DNA, even which contains CpG motifs, are taken up by antigen presenting cells via the endocytosis pathway. Endocytosed DNAs are thus degraded and their gene expression levels are inefficient. In this context, an effective plasmid delivery carrier is required for DNA vaccine development. We show in the present study that packaging plasmids containing CpGs into fusogenic liposomes (FL) derived from conventional liposomes and Sendai virus-derived active accessory proteins is an attractive method for enhancing the efficacy of a DNA vaccine. These CpG-enhanced plasmids (possessing 16 CpG repeats) that were packaged into FL, enhanced ovalbumin (OVA)-specific T cell proliferation and cytotoxic T cell activity after immunization. In fact, vaccination with CpG enhanced plasmid-loaded FL induced effective prophylactic effects compared with 13 repeats CpG containing plasmid in a tumor challenge experiment. Thus, the development of a CpG-enhanced DNA-FL genetic immunization system represents a promising tool for developing candidate vaccines against some of the more difficult infectious, parasitic, and oncologic disease targets.

Animals↗
PubMed2006

[Development of polymeric nanoparticles-based vaccine].

Traditional vaccines are mainly composed of attenuated or heat-inactivated viruses. These vaccines often generate many unwanted side effects. Subunit protein and peptide vaccines are generally very safe vaccines with well-defined components. However, proteins and peptides are often poorly immunogenic and thus require the use of adjuvants to induce adequate immunity. Therefore, particulate adjuvants have been widely investigated in vaccine delivery systems. In particular, polymeric nanoparticles with entrapped antigens represent an exciting approach to control the release of vaccine antigens and optimize the desired immune response via selective targeting of antigen to professional antigen presenting cells. It is expected that biodegradable polymeric nanoparticles have great potential as carriers for systemic and mucosal vaccine delivery systems.

Antigens↗
PubMed2006

[Study on spectroscopic properties of rare earth complexes with poly (N-vinylacetamide)].

Terbium-lanthanum (or gadolinium)-poly(N-vinylacetamide) complexes were synthesized and characterized by UV-Vis absorption spectroscopy, FTIR, XPS and fluorescence spectroscopy. The results of UV-Vis, FTIR and XPS suggested that terbium and lanthanum (or gadolinium) ions were bonded to amide group of PNVA polymer. Fluorescence experiment indicated that the characteristic emission intensity of terbium ion was greatly increased and possibly sensitized by lanthanum ion (or gadolinium). Moreover, the wavelengths of terbium characteristic emissions were changed slightly. Also the characteristic emission intensity of terbium ion doped with lanthanum ion was better than that with gadolinium ion.

English Abstract↗
PubMed2005

Rapid deswelling of semi-IPNs with nanosized tracts in response to pH and temperature.

Porous linear-polymer penetrating networks (semi-IPNs) were prepared by the radical polymerization of acrylic acid (AAc) inside porous poly(N-isopropylacrylamide) hydrogels with "nano-sized tract networks (nano-tracts)." The deswelling behavior was investigated by measuring the time course of the swelling ratio change of the semi-IPNs just after the temperature or pH was changed. Although the incorporation of PAAc linear chains into the PNIPAAm hydrogel networks should facilitate deswelling, nonporous semi-IPNs showed only a slow thermoresponse under low pH conditions, and a slow pH-response under a high temperature. On the other hand, the porous semi-IPNs showed a rapid deswelling in response to either a change in pH or temperature, regardless of the environment, presumably due to the nano-tracts through which the water was rapidly released from the hydrogel without any interference by network shrinking. The novel network design of these nano-tracted semi-IPNs may be useful for high performance, dual molecule-release functions.

Acrylic Resins↗
PubMed2005

AIDS vaccine: Intranasal immunization using inactivated HIV-1-capturing core-corona type polymeric nanospheres.

Polymeric nanospheres have been widely used in biomedical applications, such as drug, gene and vaccine delivery systems. Nanospheres with entrapped antigens have recently been shown to possess significant potential as vaccine delivery systems and adjuvants. We previously reported that concanavalin A-immobilized polystyrene nanospheres (Con A-NS) could efficiently capture HIV-1 particles and intranasal immunization with inactivated HIV-1-capturing nanospheres (HIV-NS) induced vaginal anti-HIV-1 IgA antibody responses in mice. In addition, vaginal washes from intranasally immunized mice were capable of neutralizing HIV-1. Moreover, simian/human immunodeficiency virus KU-2-capturing nanospheres (SHIV-NS) immunized macaques exhibited partial protection when vaginally and systemically challenged with pathogenic viruses. HIV-NS is suggested to be particularly suitable to enhance antigen delivery to dendritic cells (DCs). In this study, we investigated the mucosal antibody response in mice after the intravaginal or intranasal immunization in detail with using different sized (360, 660, 940 and 1230 nm) HIV-NS. The amount of immobilized Con A to NS was dependent on the surface area of the particle. Moreover, Con A-NS with different sizes could equally capture inactivated HIV-1. Intravaginal or intranasal immunization by HIV-NS with diameters ranging 360 to 1230 nm significantly induced vaginal antibody responses. However, significant differences on vaginal anti-HIV-1 gp120 IgA and IgG antibodies were not found after intravaginal or intranasal immunization with different sized HIV-NS. These results suggest that HIV-NS provides an efficient vaccine delivery system for the induction of a mucosal immune response and the development of a mucosal vaccine.

AIDS Vaccines↗
PubMed2005

Reversible thermoresponsive aggregation/deaggregation of water-dispersed polymeric nanospheres exhibiting structural transformation.

Crystalline polymeric nanospheres composed of poly{stearyl methacrylate (SMA)-co-poly(ethylene glycol) monomethacrylate (PEGm)}s were prepared by the dispersion radical polymerization of SMA and PEGm in an ethanol/water solution. Scanning electron microscopy showed that the nanospheres were highly spherical, and had a narrow size distribution. Electron spectroscopy for chemical analysis and X-ray diffraction studies of the nanospheres suggested a core-corona-type structure; the hydrophilic PEGm corona accumulated on the nanosphere surface, while the hydrophobic SMA core formed a layered structure. Heat treatment caused a melting of the SMA layers, but successive cooling allowed it to re-form. Accompanying this reversible order-disorder transition, the nanospheres also showed a reversible aggregation/deaggregation behavior in their water-dispersion state.

Journal Article↗