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Antibacterial materials are widely used to prevent hospital-acquired infections. In our previous report, metal (calcium, copper or zinc)-doped raw silk fabrics were shown to possess strong antibacterial activities against Escherichia coli. However, antibacterial materials may occasionally be harmful to the human body; thus, in this study, we investigated the cytotoxicities of extracts from metal-doped raw silk fabrics with respect to fibroblasts and osteoblasts indirectly. Calcium-doped raw silk fabric demonstrated cytocompatibility with fibroblasts. Contrarily, copper- and zinc-doped raw silk fabrics remarkably decreased the cell densities of fibroblasts, indicating their cytotoxic effects. This observation could be attributed to the high concentrations of the released copper or zinc ions. However, calcium-, copper- and zinc-doped raw silk fabrics did not demonstrate any cytotoxic effects on osteoblasts because a high concentration of the serum alleviated the effects of these metal ions released from the fabrics. Thus, calcium-doped raw silk fabric is a promising antibacterial material that does not induce strong cytotoxicity. This study will facilitate the design of materials that are both antibacterial and safe.

In the present work, the formation of zinc whitlockite via a dissolution–precipitation process was investigated in detail. The influence of medium pH, reaction time, temperature and concentration of precursors on the formation of the material was studied.

Octacalcium phosphate (OCP) has received considerable attention in the field of ceramic biomaterials as an advanced functional material. It exhibits a layered structure composed of apatitic and hydrated layers and can incorporate various dicarboxylate ions into the hydrated layer. Saturated dicarboxylic acids (HOOC(CH2)nCOOH) with an odd number of methylene groups (–CH2–) exhibit lower incorporation fractions than those with an even number of methylene groups, possibly owing to a compositional dependence on the synthetic method. In this study, calcium carbonate, phosphoric acid, and various amounts of glutaric acid were used to produce glutarate-ion-incorporated OCP by a wet chemical method, which is different from the conventional synthetic strategy. While utilising 1–20 mmol of glutaric acid during synthesis did not produce the desired product, using 25 mmol of glutaric acid resulted in the formation of single-phase glutarate-ion-incorporated OCP with a Ca/P molar ratio of 1.57 and a 90% incorporation fraction of glutarate ions. This glutarate-ion-incorporation fraction is significantly higher than that reported in the previous studies (35%). Thus, the synthetic procedure proposed herein was able to produce single-phase OCP containing glutarate ions with a high incorporation fraction. Our findings can contribute to development of novel functional ceramic biomaterials in the future.

We performed proteomic analysis of rat serum proteins adsorbed on hydroxyapatite (HAp) and α-alumina (α-Al2O3) in order to identify proteins that specifically adsorb onto HAp and control cellular responses. Proteins with either or both molecular weight of 22-32 kDa and computed isoelectric point of 5.0-5.5 were preferentially adsorbed on HAp. In total, 182 proteins were adsorbed on both HAp and α-Al2O3, of which 14 were highly enriched on HAp, whereas 68 were adsorbed only on HAp. Therefore, 82 (14+68) proteins were further evaluated by bioinformatics and literature-based analyses. We predicted that hepatocyte growth factor and angiopoietin-like protein 3 (ANGPTL3) are candidate proteins responsible for the osteoconductivity of HAp. Although ANGPTL3 promoted the attachment and spreading of MC3T3-E1 cells, it did not promote their proliferation and differentiation. Our results suggest that specific adsorption of ANGPTL3 on HAp induced osteoconductivity by enhancing the attachment and spreading of osteoblasts.

In this study, we proposed Cu-doped TiO2, prepared by the alkaline and heat treatment of titanium (Ti) metal, for use in dental applications because of its bone-bonding and antibacterial properties generated by the release of Cu ions and visible-light-responsive photocatalysis. We successfully prepared Cu-doped TiO2 on a Ti chip surface with a Cu content of 7 at.%. The apatite-forming ability of the Cu-doped TiO2 was evaluated using a simulated body fluid and it was found that apatite formation occurred. Hence, we concluded that Cu-doped TiO2 exhibits bone-bonding properties. The antibacterial properties of Cu-doped TiO2 for Escherichia coli were higher than those of non-doped TiO2 under visible light irradiation. The enhanced antibacterial effect was mainly caused by the visible-light-responsive photocatalysis of Cu-doped TiO2. We confirmed that the reactive oxygen species generated by the visible-light-responsive photocatalysis of Cu-doped TiO2 were hydroxyl radicals formed by the reaction of hydroxide ions (OH−) and holes (h+). Our findings are useful for the development of novel bioactive TiO2-based coatings and bulk materials with antibacterial properties by Cu doping.

Poly (methyl methacrylate) (PMMA) bone cement is widely used as a filler for repairing bone defects. To improve the effectiveness of the treatment for bone defects caused by metastatic bone tumours, we propose the formulation of PMMA cement containing titania (TiO2) and magnetite (Fe3O4) that offers high bone affinity, making the cement suitable for use in magnetic hyperthermia. The TiO2 and Fe3O4 contents of the PMMA cement varied from 20 to 45 mass%. The various cement samples were evaluated for their apatite-forming ability and heat-generation characteristics. The samples containing TiO2 in concentrations of 15 mass% or higher formed apatite on their surfaces within 14 days in a simulated body fluid. The heat-generation characteristics of the samples were evaluated by applying an alternating current (AC) magnetic field under the following conditions: |H| = 40 Oe and f = 600 kHz, or |H| = 100 Oe and f = 100 kHz. The surface temperatures of the samples containing 25 and 30 mass% Fe3O4 reached 42.3 and 44.8 °C, respectively, at |H| = 40 Oe and f = 600 kHz. During hyperthermia treatment, cancer cells die at temperatures higher than 42 °C, and the cement samples fabricated in this study could reach this temperature. However, since some degree of heat loss will occur in vivo, it is necessary to ensure that the temperature is higher than 42 °C by varying the AC magnetic field. Nevertheless, the fact that the samples containing Fe3O4 concentrations of 25 mass% or higher generated enough heat under the AC magnetic field makes them suitable for clinical use in hyperthermia. Thus, PMMA cement containing 15 mass% or more of TiO2 and 25 mass% or more of Fe3O4 should be investigated as a bioactive bone cement with a strong hyperthermia effect.

Carbonate-containing hydroxyapatite (CO3-HAp) shows higher osteoconductivity than pure hydroxyapatite but the underlying mechanism is unclear. As specific adsorption of proteins on CO3-HAp may affect cellular responses and osteoconductivity, we examined the contribution of protein adsorption to the high osteoconductivity observed with CO3-HAp. We prepared CO3-HAp powders with different carbonate contents (0.3, 4.8, or 8.3 wt%) and investigated the adsorption behaviour of fibronectin (Fn) on the powders. We found that the Fn adsorption capacity of CO3-HAp decreased with increasing carbonate content. Although the Hill or Langmuir isotherm was the best-fit for the adsorption of Fn onto CO3-HAp with a carbonate content of 0.3 or 4.8 wt%, the Brunauer–Emmett–Teller isotherm was the best-fit for Fn adsorption on CO3-HAp with an 8.3 wt% carbonate content. These results show that the carbonate content of CO3-HAp influences the adsorption behaviour of Fn and suggest that its specific adsorption contributes to the high osteoconductivity of CO3-HAp. The findings do not affect the clinical application of CO3-HAp directly, but they might lead to the development of bone substitutes with high osteoconductivity.

For the elucidation of the mechanism of calcium phosphate formation on commercially pure titanium (CP Ti) in the human body, rutile TiO2 single crystal plates with (001), (110), and (111) facets, n...

This study aimed to investigate extensively the full phase formation mechanism from the lowest temperature to form the phases to the optimum temperature to crystallize akermanite. The effects of various milling speeds and sintering temperatures on physico-mechanical properties of akermanite prepared using high-energy planetary milling method were also investigated. The minimum formation temperature of akermanite phase (above 800°C) was determined by X-Ray diffraction (XRD) and differential thermal analysis. XRD analysis revealed akermanite had formed through gradual phase development with the increase in temperature. Below 700C, akermanite was structurally unstable while multiple transient compounds (low clinoenstatite, wollastonite, monticellite, and diopside) coexisted, as indicated by low peak intensities. Single phase akermanite was obtained by heat-treating at 1100C. Physical studies suggested the densest akermanite ceramic feature, with tensile strength range of 25.26 ± 1.41 MPa32.10 ± 2.13 MPa and Vickers microhardness range of 1.39 ± 0.04 GPa4.94 ± 0.26 GPa could be obtained at 1250°C.

This paper reports on the influence of substituting barium (Ba2+) ions on the structural, mechanical and in vitro biological properties of akermanite prepared via high-energy wet planetary ball milling and sintering. The results obtained showed that the properties of the prepared bioceramics are highly dependent on Ba2+ content. It was found that Ba2+ resulted in improved densification (≥90%), and better apatite-forming ability of akermanite ceramic, without altering phase composition. Herein, the enhancement in diametral tensile strength (DTS), Vickers microhardness (HV) and fracture toughness (KIC) for Ba2+-doped akermanite bioceramics are reported for the first time. It was also found that after 21 days of immersion in simulated body fluid (SBF) solution, the doped ceramic surface was completely covered with apatite crystals, indicating Ba2+ encouraged apatite crystals deposition. Therefore, Ba2+-doped akermanite bioceramic could be an attractive alternative bioresorbable material for bone tissue engineering applications.

Hyperthermia treatment using appropriate magnetic materials in an alternating magnetic field to generate heat has been recently proposed as a low-invasive cancer treatment method. Magnetite (Fe3O4) nanoparticles are expected to be an appropriate type of magnetic thermal seed for this purpose, and the addition of organic substances during the synthesis process has been studied for controlling particle size and improving biological functions. However, the role of the properties of the organic polymer chosen as the modifier in the physical properties of the thermal seed has not yet been comprehensively revealed. Therefore, this study clarifies these points in terms of the molecular weight and the charge of the functional groups of the added polymers. Excepting polyethyleneimine, the Fe3O4 crystallite size decreased with increasing polymer molecular weight. Neutral polymers did not suppress the Fe3O4 formation regardless of the difference in molecular weight, while suppression of the Fe3O4 formation was observed for low molecular weight anionic polymers and high molecular weight cationic polymers. Samples with a small amount of Fe3O4 or with crystallite size less than 10 nm induced low heat generation under an alternating magnetic field.

Octacalcium phosphate (OCP), which has a layered structure composed of an apatitic layer and a hydrated layer, is a precursor of hydroxyapatite (HAp). Although the topotactic transformation of OCP ...

Ceramic biomaterials that promote bone tissue regeneration are vital in the treatment of various bone-degenerative diseases and have stimulated intense bioceramic-related research. Salts of the calcium ion and phosphate esters (SCPE) can be transformed into calcium phosphate in aqueous environments containing alkaline phosphatase (ALP). Inspired by the formation process of hydroxyapatite (HAp) in our bones, these SCPEs are promising novel ceramic biomaterials showing a biomolecule-responsive function for bone repairing applications, because our bone tissues contain ALP. In this study, the behaviour of calcium phenylphosphate (CaPP), a typical SCPE, under conditions mimicking an in vivo environment were investigated. A simulated body fluid (SBF) modified with ALP was used as reaction media. The transformation from CaPP to HAp in the SBF containing ALP occurred, while no reaction occurred without ALP present. The transformation process of CaPP can be concluded as follows: first, soaked CaPP dissolves into the SBF and forms calcium ions and phenylphosphate ions. Subsequently, phosphate ions are generated by hydrolysis of phenylphosphate ions. This hydrolytic reaction is mediated by ALP. Finally, HAp is formed by the reaction of calcium, phosphate, and hydroxide ions in the SBF. Interestingly, the transformation reaction of CaPP occurred under conditions outside the optimal pH ranges for ALP. The findings of this study demonstrate that SCPEs are good candidates for a new class of ceramic biomaterials that respond to trigger biomolecules, namely ALP, in this case.

Iron nitrides (FexNy) have attracted considerable attention as candidates for rare-earth-free hard magnetic material, and recently, the synthesis methods for FexNy nanoparticles have been established. Further, its magnetic properties might be useful for magnetic hyperthermia. Therefore, to assess its utility, the synthesis of FexNy nanoparticles was attempted in this study, and their magnetic properties and heat generation were compared with magnetite nanoparticles (MNPs), which have long been studied as biomaterials. Fe16N2 nanoparticles were successfully synthesized by reducing and nitriding MNPs with a size on the order of a few dozen nanometers, and the sample showed higher estimate heat generation than the MNPs. It was suggested that Fe16N2 nanoparticles might be used as thermal seeds for magnetic hyperthermia with higher heating efficiency.

Dense iron-doped akermanite ceramics with 0.3, 0.6 and 0.9 mol% of Fe

Neutron capture therapy, such as boron neutron capture therapy (BNCT) and gadolinium neutron capture therapy (GdNCT), has been attracting much attention as a next-generation radiotherapy. Nanoparticles containing both boron and gadolinium can accumulate in tumours because of the enhanced permeability and retention (EPR) effect, thereby simultaneously rendering the benefits of BNCT and GdNCT. In this study, we obtained flake-shaped gadolinium borate (GdBO3) nanoparticles, approximately 80 nm in length and 20 nm in width, by hydrothermal treatment of a solution containing gadolinium chloride, boric acid, and polyethylene glycol at 220 °C for 24 h. These GdBO3 nanoparticles did not show any serious cytotoxic effect against human umbilical vein endothelial cells over a concentration range of 3–300 µg·mL–1. The results suggest that GdBO3 nanoparticles can accumulate in tumours because of the EPR effect and can function as boron and gadolinium nanocarriers for neutron capture therapy. However, further in vitro and in vivo studies are needed for the application of these nanoparticles in neutron capture therapy.

Preparing drug-eluting beads (DEBs) from radiopaque materials such as titania (TiO2) can meet clinical need for directly visualizing DEBs during drug-eluting bead transcatheter arterial chemoembolization (DEB-TACE). Porous anatase-type TiO2 microspheres with mean volume diameters of approximately 30 µm were obtained when silica nanoparticles (SiNPs) were introduced into the TiO2 matrix by a sol–gel process involving a water-in-oil emulsion, and the SiNPs were then dissolved by subsequent treatment with NaOH solution. Of special note, microspheres prepared using SiNPs of approximately 20 – 25 nm in diameter had a high specific surface area of ~120 m2·g−1 and a high doxorubicin (DOX)-adsorption capacity of ~150 mg·mL−1, and they gradually released ~10% of the adsorbed DOX within 5 days. The DOX-loaded microspheres were non-cytotoxic, moreover, and exerted anticancer effects on HeLa cells. We propose that the present TiO2 microspheres are potentially useful as novel radiopaque embolic materials for DEB-TACE.

Magnetic iron oxides such as magnetite and γ-hematite have attracted considerable attention as thermoseeds for hyperthermia treatment because of their ability to generate heat under an alternating magnetic field. Control of the particle size and their combination with biocompatible polymers are expected to be beneficial for optimization of the nanoparticles. These processes can be accomplished through the synthesis of magnetite in gels, as the network structure of the polymer gel can control the grain growth of the magnetite. However, the effect of the cross-linking density of the gels remains unclear. In this study, we synthesized magnetic iron oxides in situ in chitosan hydrogels with different cross-linking densities and examined the crystalline structure and heat generation under alternating magnetic field. The crystalline phase and amount of magnetite were observed to be dependent on the cross-linking density of the gel, and the heat generation of the nanoparticles was governed by their crystalline structure and particle size rather than solely the amount of formed iron oxide.

Previous reports of some studies have described that nitrogen (N)-doped titanium dioxide (TiO2 ) exhibits photocatalytic antibacterial activity under visible light irradiation and that reactive oxygen species (ROS) is involved in its activity. For prevention and treatment of peri-implantitis, an inflammatory lesion caused by the bacterial infection of plaque adhering to the circumference of an implant, we considered that applying N-doped TiO2 to dental implant surfaces can be effective. For this study, we aimed at evaluating visible light-induced antibacterial activity of titanium (Ti) treated with NaOH and hot water, and subsequently heated in an ammonia (NH3 ) gas atmosphere at 500°C for 3 hr to quantify the generated amount of ROS available for antibacterial activity. N-doped anatase-type titania (TiO2 -xNx) is formed on the Ti substrate surface. Under visible light, markedly more hydroxyl radicals were generated with a nitrogen-doped titanium dioxide plate than with a pure titanium plate. Hydrogen peroxide exhibited the same tendency. Furthermore, it showed visible light-induced antibacterial effects over Escherichia coli. Results demonstrate that N-doped TiO2 can be useful as a dental implant surface with low risk of postoperative infection when using visible light irradiation.

Nanocomposites of magnetite (Fe3O4) and reduced graphene oxide (rGO) generate heat under an alternating magnetic field and therefore have potential applications as thermoseeds for cancer hyperthermia treatment. However, the properties of such nanocomposites as biomaterials have not been sufficiently well characterized. In this study, the osteoconductivity of Fe3O4-rGO nanocomposites of various compositions was evaluated in vitro in terms of their apatite-forming ability in simulated body fluid (SBF). Furthermore, the heat generation of the nanocomposites was measured under an alternating magnetic field. The apatite-forming ability in SBF improved as the Fe3O4 content in the nanocomposite was increased. As the Fe3O4 content was increased, the nanocomposite not only rapidly raised the surrounding temperature to approximately 100 °C, but the specific absorption rate also increased. We assumed that the ionic interaction between the Fe3O4 and rGO was enhanced and that Brown relaxation was suppressed as the proportion of rGO in the nanocomposite was increased. Consequently, a high content of Fe3O4 in the nanocomposite was effective for improving both the osteoconductivity and heat generation characteristics for hyperthermia applications.

Octacalcium phosphate (OCP) can incorporate various dicarboxylate ions in the interlayer spaces of its layered structure. Although not proven, these incorporated ions are believed to have a linear structure. In this study, the steric structures of twelve different dicarboxylate ions incorporated into OCP were investigated by comparing the experimentally determined interlayer distance of the OCP with the distance estimated using the molecular sizes of dicarboxylic acids calculated by considering their steric structures. The results revealed that the incorporated succinate, glutarate, adipate, pimelate, suberate, and aspartate ions possessed linear structures, whereas the incorporated azelate, sebacate, methylsuccinate, and malate ions exhibited bent structures. Further, the incorporated mercaptosuccinate ions featured linear, bent, other types of structures. Moreover, the steric structure of the incorporated malonate ion significantly differed from those of other dicarboxylate ions. The computational approach employed in this study is expected to deepen our understanding of the steric structures of dicarboxylate ions incorporated in the OCP interlayer spaces.

Magnetic nanoparticles are widely studied for their use in various therapeutic and diagnostic purposes. As biomaterials, their biocompatibility is as important as their magnetic properties. Iron nitride (Fex Ny ) has excellent magnetic properties, and thus Fex Ny nanoparticles could be useful as potential biomaterials. However, the biocompatibility of Fex Ny nanoparticles is yet to be investigated. In this study, we assessed the biocompatibility of Fex Ny nanoparticles by evaluating their direct-contact cytotoxicity compared with that of magnetite nanoparticles (MNPs). Rat fibroblasts were incubated with the nanoparticle samples dispersed in culture medium at concentrations of 10, 50, and 100 μg/ml. The DNA concentration measurement, MTT assay, and trypan blue exclusion test were conducted after days 1 and 3 of incubation. After day 1, the cell viability decreased, and cell death increased with increasing sample concentration when compared with the control. However, after day 3, there were no significant differences when compared with the control, irrespective of the sample concentrations. Further, there were no significant differences between the Fex Ny nanoparticles and MNPs at the same concentrations in all the cytotoxicity evaluation tests. Therefore, it is suggested that Fex Ny nanoparticles might be as cytocompatible as the conventional MNPs.

Artificial hydroxyapatite (HAp) is osteoconductive, but the mechanism is still unclear. It is likely that some serum proteins are adsorbed onto HAp and influence its osteoconductivity. We investigated the adsorption behavior of laminin (LN), which was isolated from murine Engelbreth–Holm–Swarm sarcoma, onto HAp and compared it with nonosteoconductive alpha-type alumina (α-Al2O3). Cell adhesion, spreading, and proliferation on native and LN-adsorbed discs of HAp or α-Al2O3 were examined using murine MC3T3-E1 osteoblastic cells. A larger amount of LN adsorbed onto HAp than α-Al2O3 despite the electrostatic repulsion between LN and HAp, suggesting the specific adsorption of LN onto HAp. The LN adsorbed onto HAp remarkably enhanced initial attachment and spreading of MC3T3-E1 cells, but subsequent proliferation of MC3T3-E1 cells was influenced by the type of material rather than LN adsorption. These fundamental findings imply that LN adsorbed on HAp could trigger osteoconductivity in vivo, aiding in the development of novel biomaterials that specifically adsorb LN and effectively enhance cell attachment and spreading.

Octacalcium phosphate (OCP; Ca8(HPO4)2(PO4)4 ∙ 5H2O) is a precursor of hydroxyapatite found in human bones and teeth, and is among the inorganic substances critical for hard tissue formation and regeneration in the human body. OCP has a layered structure and can incorporate carboxylate ions into its interlayers. However, studies involving the incorporation of tetracarboxylic and multivalent (pentavalent and above) carboxylic acids into OCP have not yet been reported. In this study, we investigate the incorporation of pyromellitic acid (1,2,4,5-benzenetetracarboxylic acid), a type of tetracarboxylic acid, into OCP. We established that pyromellitate ions could be incorporated into OCP by a wet chemical method using an acetate buffer solution containing pyromellitic acid. The derived OCP showed a brilliant blue emission under UV light owing to the incorporated pyromellitate ions. Incorporation of a carboxylic acid into OCP imparted new functions, which could enable the development of novel functional materials for biomedical applications. Octacalcium phosphate is a precursor to a key component of human bone and tooth enamel which can incorporate carboxylate species, but the effect of multivalent carboxylates is not well understood. Here the incorporation of tetracarboxylic acids into OCP is shown to influence the structural and optical properties of the resultant composite.

Raw silk has the potential to be a flexible, osteoconductive material because it forms bone-like apatite on its surface in acellular simulated body fluid with ion concentrations nearly 1.5 times greater than that of human plasma (1.5SBF). It has been reported that silk—which has many similarities to raw silk—develops antibacterial properties when heated in inert gas, which may be advantageous in preventing bacterial infection. Hence, raw silk heated in inert gas may be a flexible, osteoconductive material with antibacterial activity. Thus, we examined the effect of the heat treatment of raw silk fabric on its apatite-forming ability in 1.5SBF and on the growth of Escherichia coli. Raw silk fabric was heated in argon gas at several temperatures, to a maximum of 500 °C. The results of soaking tests in 1.5SBF indicate that the apatite-forming ability of raw silk decreases with increasing temperature. This may be because favourable structures for apatite formation, such as carboxyl groups, are thermally decomposed. The results of bacterial tests indicate that raw silk fabrics heated to 300 °C or 500 °C exhibit reduced bacterial growth compared to those that were not heated or were heated only to 100 °C. This might be because hydrophobic surfaces inhibit bacterial adhesion, or because the thermal decomposition of sericin—a component of raw silk—leads to a lack of available nutrients for the bacteria. Although this study did not demonstrate the expected material properties needed for clinical applications, this research contributes to a better understanding of silk biomaterials.