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Biomedical subjects

Fumio Watari

Publications and source records attributed to Fumio Watari.

At least 19 recordsLinked to original sources

Mechanical properties and biological behavior of carbon nanotube/polycarbosilane composites for implant materials.

Multiwalled carbon nanotube/polycarbosilane (MWCNT/PCS) composites were fabricated by the spark plasma sintering (SPS) method. The MWCNT/PCS composites consisted of MWCNTs and nanosized SiC particles pyrolyzed from PCS and possessing good mechanical properties for bone tissue repair or dental implantation. The MWCNT/PCS composites were implanted in the subcutaneous tissue and femur of rats at 1 and 4 weeks after implantation. Histological investigations showed that there was little inflammatory response in the subcutaneous tissue, and newly formed bone tissue was observed in the femur. These results indicated that the MWCNT/PCS composite had little prophlogistic effect and good osteoconductivity. The study suggested the possibility that the MWCNT/PCS composite could be a candidate bone-substitute and dental-implant material in the future.

Animals↗

The degradation of the three layered nano-carbonated hydroxyapatite/collagen/PLGA composite membrane in vitro.

OBJECTIVE: The purpose of this paper was to investigate the in vitro biodegradation of a guided tissue regeneration composite membrane, nano-carbonated hydroxyapatite/collagen/poly(lactic-co-glycolic acid) (nCHAC/PLGA). Especially for periodontal therapy, the functional graded material (FGM) nCHAC/PLGA membrane was prepared that consisted of three layers with 8 wt% nCHAC + PLGA/4 wt% nCHAC + PLGA/PLGA, where one face of the membrane is porous, thereby allowing cell growth thereon and the opposite face of the membrane smooth, thereby inhibiting cell adhesion. METHODS: For evaluation, in vitro degradation specimens of nCHAC/PLGA were immersed into artificial saliva solution at 37 degrees C for 1, 2, 4, 8 and 12 weeks to detect the weight loss over the period, and set pure PLGA membrane as control to compare the degraded behaviors. pH value and calcium concentration of the residual solution were measured, and morphology change was investigated by scanning electron microscopy (SEM). RESULTS: During the experimental period in vitro, the whole shape of the membrane could be kept for 4 weeks, after that it became powder at between 8 and 12 weeks. The results demonstrated that weight loss increased continuously with a reduction in mass of 23.1% after 4 weeks and 88% after 12 week for the nCHAC/PLGA three FGM layers composite membrane. The calcium concentration in the residual solution showed a significant increase after 4 weeks, which referred to the nano-carbonated hydroxyapatite degradation. Moreover, the pH value in the solution of the nCHAC/PLGA membrane was a little higher than that of the pure PLGA membrane, which demonstrated the possible neutralization effect from nCHAC composite for the acid outcome of PLGA in the solution. The pore structure of 8 wt% nCHAC + PLGA was enlarged on the porous surface, while the nonporous surface of pure PLGA also showed a small porous structure after increased time. SIGNIFICANCE: Degradation of the composite membrane is appropriate for practical periodontal repair. Moreover, the new mineral formation on the surface of the composite membrane referred to the possible positive effect in vivo for new bone tissue regeneration.

Absorbable Implants↗

Human neutrophils reaction to the biodegraded nano-hydroxyapatite/collagen and nano-hydroxyapatite/collagen/poly(L-lactic acid) composites.

The impact of biodegraded nano-hydroxyapatite/collagen (nHAC) composite and nano-hydroxyapatite/collagen/poly(L-lactic acid) (nHAC/PLA) scaffold composite on neutrophils reaction was evaluated in vitro. Neutrophils were separated from human peripheral blood of healthy subjects. The nHAC and nHAC/PLA materials were immersed in the D-Hanks' Balanced Salt Solution (D-HBSS) for 1 day, 7 days and 2, 4, 8 weeks (37 degrees C) as testing solution, which mixed with the neutrophils for 1 h. Both of the nHAC and nHAC/PLA materials were shown the same cell survival rate as blank control, but the lactate dehydrogenase (LDH) and tumor necrosis factor alpha (TNF-alpha) released from the neutrophils were increased significantly after the 2 weeks in nHAC sample. The possible reason relied on the high concentration of calcium due to the quick biodegradation of the nHAC material. Before 2 weeks, the LDH value of nHAC/PLA is higher than that of nHAC sample that corresponded to the initial PLA degradation in vitro. This study provided the biocompatibility test of neutrophils other than common methods, such as osteoblastic cells for biomimetic materials. Moreover, it demonstrated the calcium concentration stimulating effect for cytokine release from neutrophils.

Biocompatible Materials↗

Effect of surface condition of dental zirconia ceramic (Denzir) on bonding.

Yttria partially stabilized zirconia (YPSZ) ceramics are suitable for dental and medical use because of their high fracture toughness and chemical durability. The purpose of this study was to examine the bonding behavior of a dental YPSZ ceramic, Denzir. After being subjected to various surface treatments, Denzir specimens were bonded to each other using an adhesive resin composite, glass ionomer, or zinc phosphate cement. Bonding strength was then determined by the shearing test. No significant differences (p>0.05) were observed between SiC- and Al2O3-blasted specimens. In all surface treatments, the shear bond strength significantly (p<0.05) increased in the order of adhesive resin composite cement > glass ionomer cement > zinc phosphate cement. Moreover, silanization with methacryloxy propyl trimethoxysilane slightly increased the bonding strength of the adhesive resin composite cement.

Dental Bonding↗

In situ quantitative analysis of etching process of human teeth by atomic force microscopy.

Etching is one of the most fundamental steps in the restoration of teeth by adhesion of composite resin in dental clinics. Atomic force microscope (AFM) was used for the in situ observation of the etching process of human enamel, dentin and synthetic hydroxyapatite in the three different acid agents, 2% phosphoric acid, 10% citric acid and 10% polyacrylic acid. To measure the absolute depth from the initial level before etching and to correlate the surface height between the changing AFM images obtained, the depth profiles were recorded with etching time by carrying out the line scan consecutively at the representative place of the observed area. These chronological series of depth profiles enabled us to perform quantitative analysis of etched amount in addition to the surface roughness obtained from relative depth profile within one image. The course of etching process from the dissolution of smear layer, appearance of enamel prizms or dentinal tubules to progress of demineralization could clearly be observed. The depth profile, surface roughness, etching amount, etching rate and smear layer thickness could then be evaluated. The different etching characteristics of three acid agents and the effect of surface roughness produced by different mechanical prepolish were compared and discussed.

Acid Etching, Dental↗

Influence of length on cytotoxicity of multi-walled carbon nanotubes against human acute monocytic leukemia cell line THP-1 in vitro and subcutaneous tissue of rats in vivo.

Carbon nanotubes (CNTs) are single- or multi-cylindrical graphene structures that possess diameters of a few nanometers, while the length can be up to a few micrometers. These could have unusual toxicological properties, in that they share intermediate morphological characteristics of both fibers and nanoparticles. To date, no detailed study has been carried out to determine the effect of length on CNT cytotoxicity. In this paper, we investigated the activation of the human acute monocytic leukemia cell line THP-1 in vitro and the response in subcutaneous tissue in vivo to CNTs of different lengths. We used 220 nm and 825 nm-long CNT samples for testing, referred to as "220-CNTs" and "825-CNTs", respectively. 220-CNTs and 825-CNTs induced human monocytes in vitro, although the activity was significantly lower than that of microbial lipopeptide and lipopolysaccharide, and no activity appeared following variation in the length of CNTs. On the other hand, the degree of inflammatory response in subcutaneous tissue in rats around the 220-CNTs was slight in comparison with that around the 825-CNTs. These results indicated that the degree of inflammation around 825-CNTs was stronger than that around 220-CNTs since macrophages could envelop 220-CNTs more readily than 825-CNTs. However, no severe inflammatory response such as necrosis, degeneration or neutrophil infiltration in vivo was observed around both CNTs examined throughout the experimental period.

Animals↗

Strict preparation and evaluation of water-soluble hat-stacked carbon nanofibers for biomedical application and their high biocompatibility: influence of nanofiber-surface functional groups on cytotoxicity.

Water-soluble H-CNFs modified with a carboxyl group possessed the ability to induce TNF-alpha, whereas CHAPS-treated H-CNFs possessed significantly greater activity and were also found to activate NF-kappaB reporter activity, to a significantly greater level than H-CNFs; furthermore the functional group modified or coated on the surface of H-CNFs was a significant cytotoxic factor that affected cell activation.

Carbon↗

The preparation and characteristics of a carbonated hydroxyapatite/collagen composite at room temperature.

Nanocarbonated hydroxyapatite/collagen (nCHAC) composite was prepared at room temperature via biomimetic self-assembly method. X-ray diffraction (XRD), thermogravimetric analysis (TGA), and transmission electron microscopy (TEM) were performed. This composite shows the same inorganic phase of natural bone with nanosized level and low degree of crystallinity, and contains 2.8-14.7 wt % of carbonated content. TEM results confirm that the microstructure of this composite is the mineralized collagen fiber bundle like the hierarchical structure of natural bone. The diameter of a single mineralized collagen fiber is about 4 nm. Slightly different assembly units of the composite with different carbonates and collagen were demonstrated. The carbonated percentage affects the mineral crystal size and collagen fibril assembly. Because of the biomimetic component and microstructure, the use of nCHAC composite is promising for hard tissue therapy.

Bone Substitutes↗

Biomimetic porous scaffolds with high elasticity made from mineralized collagen--an animal study.

Histological investigations of a new hydroxyapatite-collagen composite material were carried out to evaluate its possible suitability as a bone substitute. The three-dimensional scaffolds made from biomimetically mineralized collagen exhibit an interconnecting pore structure and elastic mechanical properties. They were implanted into the subcutaneous tissue and bone defects made in the femur of rats and harvested with the surrounding tissue at 1, 2, 4, 8, and 12 weeks after surgery. The materials implanted in the subcutaneous tissue were covered by fibrous connective tissue with a slight inflammatory response, and many foreign-body giant cells were observed on the surface of the scaffolds. Most of the material implanted in the subcutaneous tissue was resorbed at 8 weeks by phagocytosis. In the bone defects, new bone formation was observed on the surface of the material at 1 week. New bone increased with time, and osteoclasts were seen on the surface of the scaffolds at 2 weeks. Resorption and replacement by new bone of many parts of the materials implanted in the femur were observed by 12 weeks. These responses occurred faster than those of other hydroxyapatite-collagen composites. The results suggested that the new biomimetically mineralized collagen scaffolds were suitable as an implant material for bone-tissue reconstruction.

Animals↗

A three-layered nano-carbonated hydroxyapatite/collagen/PLGA composite membrane for guided tissue regeneration.

Functional graded materials (FGM) provided us one new concept for guided tissue regeneration (GTR) membrane design with graded component and graded structure where one face of the membrane is porous thereby allowing cell growth thereon and the opposite face of the membrane is smooth, thereby inhibiting cell adhesion in periodontal therapy. The goal of the present study was to develop a three-layered graded membrane, with one face of 8% nano-carbonated hydroxyapatite/collagen/poly(lactic-co-glycolic acid) (nCHAC/PLGA) porous membrane, the opposite face of pure PLGA non-porous membrane, the middle layer of 4% nCHAC/PLGA as the transition through layer-by-layer casting method. Then the three layers were combined well with each other with flexibility and enough high mechanical strength as membrane because the three layers all contained PLGA polymer that can be easily used for practical medical application. This high biocompatibility and osteoconductivity of this biodegraded composite membrane was enhanced by the nCHAC addition, for the same component and nano-level crystal size with natural bone tissue. The osteoblastic MC3T3-E1 cells were cultured on the three-layered composite membrane, the primary result shows the positive response compared with pure PLGA membrane.

Animals↗

Biological behavior of hat-stacked carbon nanofibers in the subcutaneous tissue in rats.

The tissue response to hat-stacked carbon nanofibers (H-CNFs) was evaluated. H-CNFs were implanted in the subcutaneous tissue of rats. Histological and ultrastructural investigations were carried out by transmission electron microscopy. Although many macrophages and foreign body giant cells were seen around H-CNFs, no severe inflammatory response such as necrosis was observed. Some H-CNFs were observed in lysosomal vacuoles of phagocytes. These results showed that H-CNFs were not strong prophlogistic substances and were englobed in vivo.

Animals↗

Novel bulk carbon nanotube materials for implant by spark plasma sintering.

Novel, bulk multiwall carbon nanotubes (MWCNTs) sintered with polycarbosilane (PCS) as a binder agent were fabricated by spark plasma sintering (SPS), and their microstructure and properties were investigated. Sintering was done with 20-60 MPa pressure at 1200 degrees C. SEM and TEM observations showed that the nanosized tube microstructure was preserved even after sintering, and tubes adhered to each other with the nanosized nodules of SiC pyrolyzed form PCS as revealed by X-ray diffraction. Bulk density and Vickers hardness were found to increase, whereas the specific surface area decreased, as PCS content and sintering pressure increased. Through animal experiments, the inflammatory reaction of CNTs/PCS material was found to be slightly increased with increasing PCS content. In conclusion, sintered CNTs had physical and mechanical properties close to bone, and their good biocompatibility based on tissue response served to pave their way as a suitable implant material in the future.

Animals↗

Rare earth oxide-containing fluorescent glass filler for composite resin.

In recent dental care, esthetic restorative materials such as composite resin and porcelain have been widely used and studied. However, their good esthetics makes the visual inspection of restored teeth difficult. In this study, a fluorescent glass filler containing rare earth oxides - which are well-known fluorescent materials - was prepared and used in composite resin to enhance visual inspection capability with small change in color. For example, when irradiated with near ultraviolet light, an Eu2O3-containing filler fluoresced clearly and visibly. The fluorescence intensity of the prepared composite resin increased with increase of Eu2O3 content in the glass filler and with filler/resin ratio in the resin. Despite the clear fluorescence, the effect of Eu2O3 on the color change of composite resin was quite small - even when up to 10 wt% Eu2O3 was added to the glass filler. Tb4O(7-) and Dy2O3-added fillers also showed clear fluorescence, just like Eu2O3-added filler. Therefore, fluorescent glass fillers for composite resins, resulting in small color change in the latter, were successfully prepared in this study.

Composite Resins↗

Caged multiwalled carbon nanotubes as the adsorbents for affinity-based elimination of ionic dyes.

Multiwalled carbon nanotubes (MWCNTs) were used as the active elements for the first time for affinity-based elimination of ionic dyes. MWCNTs were encapsulated in cross-linked alginate (ALG) microvesicles using Ba2+ as the bridging ion. The Ba2+-alginate matrix constitutes a cage which holds the physically trapped MWCNTs. The cage carries negative charges on its surface. The cage restricts the access of anions of large molecular weight, such as humic acids, because of electrostatic repulsion. The cage also restricts the access of colloids of large size, because of size exclusion. Ionic dyes partition into the cage and then are captured by MWCNTs probably on the basis of van der Waals interactions occurring between the hexagonally arrayed carbon atoms in the graphite sheet of MWCNTs and the aromatic backbones of the dyes. As a result of these interactions the target species, namely, the ionic dyes, are eliminated efficiently by the MWCNTs of Ba2+-ALG/MWCNT composite adsorbents. The adsorptive capacities for elimination of acridine orange, ethidium bromide, eosin bluish, and orange G (the model species used for this study) were found as high as 0.44, 0.43, 0.33, and 0.31 micromol, respectively, for 1.0 mg of the caged MWCNTs. Adsorptive experiments with carbon nanofibers and activated carbons as the adsorbents were also performed. The MWCNT-based adsorbents provided the best capability for the affinity-based elimination of these targeted species. Biocompatibility experiments performed in vitro and in vivo provided promising results, suggesting potential applications of the caged MWCNTs in in situ environmental remediation.

Adsorption↗

Three-dimensional topographic scanning electron microscope and Raman spectroscopic analyses of the irradiation effect on teeth by Nd:YAG, Er: YAG, and CO(2) lasers.

A three-dimensional analyzer installed in a scanning electron microscope was used to evaluate the morphology and surface roughness using noncontact profilometry. Observations were carried out on the enamel and dentin surface irradiated by three different lasers: Nd:YAG (wavelength 1.06 microm), Er:YAG (2.94 microm), and CO(2) (10.6 microm). Spectroscopic analysis was done by Raman spectroscopy for nonirradiated and laser-irradiated surfaces. The lasers were applied perpendicularly to vertically sectioned and polished human extracted caries-free molars. The tooth was sectioned at each cavity for cross-section analysis after laser irradiation. Irradiation by Nd:YAG and CO(2) lasers of the enamel surface showed an opaque white color, different from dentin where the surface turned black. The Er:YAG laser induced no changes in color of the dentin. Numerous cracks associated with thermal stress were observed in the CO(2) laser-irradiated dentin. Noncontact surface profile analysis of Er:YAG laser-irradiated enamel and dentin showed the deepest cavities, and direct cross-sectional observations of them showed similar cavity outlines. The CO(2) laser-irradiated dentin had the least surface roughness. Raman spectroscopic analysis showed that fluorescence from the laser-irradiated tooth was generally greater than from nonirradiated teeth. Bands in dentin attributed to organic collagen matrix were lost after Nd:YAG and CO(2) laser irradiation, and a broad peak due to amorphous carbon appeared. The Er:YAG laser-irradiated dentin showed no sign of a carbon band and had more suitable results for dental ablation. Noncontact surface profile analysis was effective to evaluate the structural change in the tooth in the microarea of study after laser irradiation.

Humans↗

Microparticle formation and its mechanism in single and double emulsion solvent evaporation.

The emulsification is the first step of the emulsification solvent evaporation method and has been extensively investigated. On the contrary the second step, the solvent transport out from the emulsion droplets that determine the particle morphology and with great influence on the microparticles encapsulation and release behavior has been scarcely studied. This study investigates the mechanism of the solvent elimination from the emulsion droplets and its influence on the particle morphology, encapsulation and release behavior. Usually, the solvent is highly volatile that makes the solvent elimination process very fast thus difficult to observe. In order to observe in detail the microparticle formation, the initial emulsion was monitored by optical microscope under controlled solvent evaporation conditions. The results from the optical microscopic observations corroborated with laser diffractometry analysis showed that in single emulsion formulations, spherical microparticles are formed by accelerated solvent elimination due to the combined effects of high solvent volatility and polymer precipitation. The solvent expulsion accompanied by important shrinkage generates on the microparticle surface a thin layer of nanoparticles attested by scanning electron microscopy and laser diffractometry. During the intense solvent elimination, the encapsulated substance is drained, affecting the loading efficiency. Furthermore, it will concentrate towards the microparticle surface contributing to the initial burst release. In double emulsion formulations, microparticles with different morphologies are generated due to the presence of the aqueous-phase microdroplets inside the emulsion droplet. During the solvent elimination, these microdroplets generally coalesce under the pressure of the precipitating polymer. Depending mainly on the polymer concentration and emulsification energies, the final microparticles will be a mixture of honeycomb, capsule or plain structure. During the shrinkage due to the incompressibility of the inner microdroplets, the precipitating polymer wall around them may break forming holes through which the encapsulated substance is partly expulsed. Through these holes, the encapsulated substance is further partitioning with the external aqueous phase during solvent evaporation and contributes to the initial burst release during the application.

Capsules↗

Quantitative analysis of biologic specimens by X-ray scanning analytic microscopy.

X-ray scanning analytic microscopy (XSAM) can be used to visualize the elemental distribution in biologic specimens. In this article, the authors prepared standard specimens for XSAM and performed quantitative analysis of various elements dissolved in soft tissues. Two different types of standard specimens were prepared. Methylmethacrylate (MMA) resin-based standard specimens were prepared with organic compounds of elements for low-concentration standards and lithium borate glass-based standard specimens were prepared with oxides of elements for higher concentration standards. Using these standard specimens, the P and Ca concentrations in normal rat tissue and dissolved Ni, Fe, and Ni concentrations around metal-implanted tissues were quantitatively analyzed. The estimated concentrations of dissolved Fe, Cu, and Ni from the implants were 1000, 40, and 20 mM, respectively. From the concentration levels causing inflammation around these implants, the high toxicity for soft tissue of Ni and Cu at low concentrations, for example, 10 mM, was confirmed. The toxicity of Cu was estimated as next to that of Ni. In contrast, Fe had low toxicity despite high concentrations of dissolved Fe of as much as 1000 mM. In this article, it was possible to estimate the nonmetallic elements and low-concentration metallic elements dispersed in soft tissue by XSAM.

Animals↗