Fabrication of quantum dot-immobilized Y2O3 microspheres with effective photoluminescence for cancer radioembolization therapy.

Microspheres composed of Y-containing materials are effective agents for cancer radioembolization therapy using ß-rays. The distribution and dynamics of these microspheres in tissues can be easily determined by providing the microspheres with an imaging function. In addition, the use of quantum dots will enable the detection of microspheres at the individual particle level with high sensitivity. In this study, core - shell quantum dots were bound to chemically modified yttria microspheres under various conditions, and the effect of reaction conditions on the photoluminescence properties of the microspheres was investigated. The quantum dots were immobilized on the surfaces of the microspheres through dehydration - condensation reactions between the carboxy groups of quantum dots and the amino groups of silane-treated microspheres. As the reaction time increased, the photoluminescence peak blue shifted, and the photoluminescence intensity and lifetime decreased. Therefore, a moderate period of the immobilization process was optimal for imparting effective photoluminescence properties. This study is expected to facilitate particle-level tracking of microsphere dynamics in biological tissues for the development of minimally invasive cancer radiotherapy of deep-seated tumors.

We have established a method to immobilize quantum dots on yttria microspheres for cancer radiotherapy and revealed that photoluminescence intensity can be optimized by controlling the immobilization treatment time.

Proliferation and differentiation of MC3T3-E1 cells on polymethyl methacrylate cements containing Fe3O4 and TiO2 for hyperthermic treatment of metastatic bone tumors.

Polymethyl methacrylate (PMMA) bone cement is widely used to relieve pain caused by metastatic bone tumors. We previously found that PMMA bone cement containing 15 mass% or more of TiO2 showed good apatite-forming ability, and 25 mass% or more of Fe3O4 generated sufficient heat for hyperthermia under an alternating current (AC) magnetic field. In this study, the cytocompatibility of PMMA bone cement with Fe3O4:TiO2 weight ratios of 25:15 (F25T15-3/2-42) and 30:15 (F30T15-3/2-42) was evaluated using osteoblastic cells (MC3T3-E1). The proliferation and differentiation of MC3T3-E1 cells were suppressed for F25T15-3/2-42 and F30T15-3/2-42 compared to PMMA bone cement without Fe3O4 and TiO2 (F0T0-3/2-42). The release of methyl methacrylate (MMA) monomers from F25T15-3/2-42 and F30T15-3/2-42 at 7 days was about 33 and 50 times higher than that from F0T0-3/2-42, respectively. The remarkable release of MMA monomers from F25T15-3/2-42 and F30T15-3/2-42 may be responsible for the suppressed proliferation and differentiation of MC3T3-E1 cells. The release of MMA monomers was not reduced when the MMA/PMMA weight ratio was decreased from 3/2 to 1/1, however, it was significantly reduced by increasing the content of benzoyl peroxide (BPO) and N, N-dimethyl-p-toluidine (DMPT) to 8 and 4 mass% against MMA, respectively. Proliferation and differentiation of MC3T3-E1 cells on PMMA-type cements containing Fe3O4 and TiO2 with increased BPO and DMPT contents need to be investigated in the future; however, our findings will be useful for designing PMMA cements for the hyperthermic treatment of metastatic bone tumors.

Tailoring mechanical and in vitro biological properties of calcium‒silicate based bioceramic through iron doping in developing future material.

Dense iron-doped akermanite ceramics with 0.3, 0.6 and 0.9 mol% of Fe3+ were synthesized via high-speed planetary ball milling and subsequently subjected to sintering at 1200 and 1250 °C. The aim of the current work was to investigate the effect of trivalent iron (Fe3+) in tuning the physicomechanical and in vitro biological properties of akermanite. The incorporation of Fe3+ into akermanite host and sintering at a high temperature of 1200 °C resulted in a synergistic effect in enhancing the sinterability and densification of akermanite ceramics. Although varying the Fe3+ content, it was found that similar densification and mechanical properties (i.e., diametral tensile strength, Vickers microhardness and fracture toughness) were observed for the doped ceramics at 1250 °C, indicating that this newly developed formulation is temperature-dependent. Fe3+-doped akermanite ceramics revealed greater in vitro bioactivity as compared to undoped akermanite, demonstrated by better coverage of needle-like apatite precipitates after 21 days of immersion in simulated body fluid. Additionally, Rat-1 cells cultured in direct contact with Fe3+-doped akermanite ceramics showed almost double levels of cell proliferation than their undoped counterpart on both 3 and 7 days of culture. Our finding suggests that 0.9Fe-AK ceramic is a suitable formulation to be considered for future bone substitute material as it provides sufficient mechanical strength as well as good bioactivity and the ability to encourage cell proliferation.

Cytotoxicity evaluation of iron nitride nanoparticles for biomedical applications.

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.

Apatite formation and bacterial growth on raw silk fabric heated in argon gas.

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.

Structural control of magnetite nanoparticles for hyperthermia by modification with organic polymers: effect of molecular weight.

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.

Visible light-induced photocatalytic and antibacterial activity of N-doped TiO2.

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.

In situ synthesis of magnetic iron oxide nanoparticles in chitosan hydrogels as a reaction field: Effect of cross-linking density.

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.

In vitro apatite mineralization and heat generation of magnetite-reduced graphene oxide nanocomposites for hyperthermia treatment.

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.

Spherical porous hydroxyapatite granules containing composites of magnetic and hydroxyapatite nanoparticles for the hyperthermia treatment of bone tumor.

Spherical porous granules of hydroxyapatite (HA) containing magnetic nanoparticles would be suitable for the hyperthermia treatment of bone tumor, because porous HA granules act as a scaffold for bone regeneration, and magnetic nanoparticles generate sufficient heat to kill tumor cells under an alternating magnetic field. Although magnetic nanoparticles are promising heat generators, their small size makes them difficult to support in porous HA ceramics. We prepared micrometer-sized composites of magnetic and HA nanoparticles, and then supported them in porous HA granules composed of rod-like particles. The spherical porous HA granules containing the composites of magnetic and HA nanoparticle were successfully prepared using a hydrothermal process without changing the crystalline phase and heat generation properties of the magnetic nanoparticles. The obtained granules generated sufficient heat for killing tumor cells under an alternating magnetic field (300 Oe at 100 kHz). The obtained granules are expected to be useful for the hyperthermia treatment of bone tumors.

Adsorption of Laminin on Hydroxyapatite and Alumina and the MC3T3-E1 Cell Response.

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.

In vitro apatite formation and drug loading/release of porous TiO2 microspheres prepared by sol-gel processing with different SiO2 nanoparticle contents.

Bioactive titania (TiO2) microparticles can be used as drug-releasing cement fillers for the chemotherapeutic treatment of metastatic bone tumors. Porous anatase-type TiO2 microspheres around 15 µm in diameter were obtained through a sol-gel process involving a water-in-oil emulsion with 30:70 SiO2/H2O weight ratio and subsequent NaOH solution treatment. The water phase consisted of methanol, titanium tetraisopropoxide, diethanolamine, SiO2 nanoparticles, and H2O, while the oil phase consisted of kerosene, Span 80, and Span 60. The resulting microspheres had a high specific surface area of 111.7 m(2)·g(-1). Apatite with a network-like surface structure formed on the surface of the microspheres within 8 days in simulated body fluid. The good apatite-forming ability of the microspheres is attributed to their porous structure and the negative zeta potential of TiO2. The release of rhodamine B, a model for a hydrophilic drug, was rapid for the first 6 h of soaking, but diffusion-controlled thereafter. The burst release in the first 6h is problematic for clinical applications; nonetheless, the present results highlight the potential of porous TiO2 microspheres as drug-releasing cement fillers able to form apatite.

Bisphosphonate release profiles from magnetite microspheres.

Hyperthermia has been suggested as a novel, minimally invasive cancer treatment method. After implantation of magnetic nano- or microparticles around a tumour through blood vessels, irradiation with alternating magnetic fields facilitates the efficient in situ hyperthermia even for deep-seated tumours. On the basis of this idea, if the microspheres are capable of delivering drugs, they could be promising multifunctional biomaterials effective for chemotherapy as well as hyperthermia. In the present study, magnetite microspheres were prepared by aggregation of the iron oxide colloid in water-in-oil (W/O) emulsion. The release behaviour of alendronate, a typical bisphosphonate, from the microspheres was examined in vitro as a model of the bone tumour prevention and treatment system. The alendronate was successfully incorporated onto the porous magnetite microspheres in vacuum conditions. The drug-loaded microspheres maintained their original spherical shapes even after shaking in ultrapure water for 3 days, suggesting that they have sufficient mechanical integrity for clinical use. It was attributed to high aggregation capability of the magnetite nanoparticles through van der Waals and weak magnetic attractions. The microspheres showed slow release of the alendronate in vitro, resulting from tight covalent or ionic interaction between the magnetite and the alendronate. The release rate was diffusion-controlled type and well controlled by the alendronate concentration in drug incorporation to the microspheres.

MC3T3-E1 and RAW264.7 cell response to hydroxyapatite and alpha-type alumina adsorbed with bovine serum albumin.

Initial cell responses following implantation are important for inducing osteoconductivity. We investigated cell adhesion, spreading, and proliferation in response to native and bovine serum albumin (BSA)-adsorbed disc of hydroxyapatite (HA) or alpha-type alumina (α-Al2O3) using mouse MC3T3-E1 osteoblastic cells and mouse RAW264.7 macrophages. The adsorbed BSA inhibited adhesion and spreading of MC3T3-E1 cells, but did not affect MC3T3-E1 cell proliferation on HA and α-Al2O3 substrates. Thus, MC3T3-E1 cells quickly adhere to original HA before cell binding is impeded by adsorption of BSA in quantities sufficient to inhibit the adhesion of MC3T3-E1 cells. The adsorbed BSA inhibits adhesion of RAW264.7 cells to α-Al2O3, but not to HA. BSA adsorption does not affect RAW264.7 cell spreading and proliferation on both HA and α-Al2O3 substrates. Thus, BSA adsorbed on HA stimulates a different cell response than α-Al2O3. Moreover, quick adherence of osteoblast cells and monocyte-macrophage lineage cells plays a role in HA osteoconductivity.

Carboxymethyldextran/magnetite hybrid microspheres designed for hyperthermia.

Recently, organic-inorganic hybrids composed of derivatives of dextran, a polysaccharide, and magnetite nanoparticles have attracted much attention as novel thermoseeds. If they can be fabricated into microspheres of size 20-30 µm, they are expected to show not only hyperthermia effects but also embolization effects in human liver and kidney cancers. In this study, we examined the fabrication of carboxymethyldextran/magnetite microspheres using a water/oil emulsion as the reaction medium. Improvement of the chemical stability of the microcapsules by coating with silica using a sol-gel process was also investigated. The obtained hollow microspheres contained particles of size 20-30 µm. Silica coating using an appropriate catalyst for hydrolysis and polycondensation of alkoxysilanes was found to be effective for preventing dissolution and collapse in simulated body environments.

Sol-gel synthesis, characterization, and in vitro compatibility of iron nanoparticle-encapsulating silica microspheres for hyperthermia in cancer therapy.

We prepared iron nanoparticle-encapsulating silica (FeSi) microspheres and tested their suitability as thermal seeds for hyperthermia in cancer therapy. These microspheres were prepared by introducing a ferric ion (Fe(3+)) into microspheres of a SiO(2) gel matrix derived from the hydrolysis of tetramethoxysilane in a water-in-oil emulsion that was then heat-treated at 850 °C in an argon atmosphere. The particles obtained were 5-30 µm in size and had a saturation magnetization up to 21 emu g(-1) and a coercive force of 86-133 Oe. Heat generation in an alternating current magnetic field of 300 Oe at 100 kHz was estimated to be 7.7-28.9 W g(-1). The in vitro cell biocompatibility of the microspheres was assessed by culturing rat fibroblast Rat-1 cells in medium supplemented with microspheres containing 6.7 % of iron nanoparticles. At microsphere concentrations of <7.5 g L(-1) proliferation of Rat-1 cells was not significantly inhibited.

In vitro assessment of poly(methylmethacrylate)-based bone cement containing magnetite nanoparticles for hyperthermia treatment of bone tumor.

Poly(methylmethacrylate) (PMMA)-based cements containing magnetite (C-PMMA/Fe(3)O(4)) is useful in hyperthermia treatment for bone tumor. We have prepared C-PMMA/Fe(3)O(4) by incorporating Fe(3) O(4) powders of different diameters (means of 300, 35, and 11 nm) into the polymerization reaction of methyl methacrylate monomer to develop a new bone cement with high heating efficiencies in alternating current (AC) magnetic fields. Further, we have investigated the in vitro heating capability of the cements in different AC magnetic fields. The mechanical strength and biocompatibility of the resultant cements were also assessed. Their heat generation strongly depends on the magnetite nanoparticle sizes and applied magnetic fields. The cement containing Fe(3)O(4) with mean diameter around 35 nm exhibited the highest heating capability in AC magnetic fields of 120 and 300 Oe at 100 kHz while that with mean diameter around 11 nm exhibited optimum heating capability in AC magnetic fields of 40 Oe at 600 kHz. The incorporation of Fe(3)O(4) into cement-30 wt % of the total amount of cement-did not significantly change the compressive strength of cement, and the proliferation of rat fibroblast Rat-1 cells on cement discs was not inhibited. Our investigations are useful for designing new PMMA/Fe(3)O(4) bone cement with high heating efficiencies and biocompatibilities for bone tumor treatments.

Current progress in inorganic artificial biomaterials.

In this review, recent advances in bioceramics, metallic biomaterials, and their composites are discussed in terms of their material properties and new medical applications. Porous calcium phosphate ceramics have attracted a lot attention as scaffolds for tissue-engineering purposes since the porous structure allows bone ingrowth. The addition of biodegradable polymers like chitosan, gelatin, and collagen have modified the degradability of the ceramics and their mechanical properties. Titanium (Ti) alloys are being developed for the fabrication of medical devices for the replacement of hard tissue such as artificial hip joints, bone plates, and dental implants because they are very reliable from the viewpoint of mechanical performance. Physical treatment such as grooving or setting a spatial gap on the surface of materials is applicable to improve the apatite formation on the Ti alloys. Blood-compatible polymers such as poly(ethylene glycol) have been successfully fixed on the surface of Ti via chemical bonding by an electrodeposition method. New functions have been explored in Ni-free, Co-Cr-Mo alloys and Mg alloys. In addition, yttrium-containing or phosphorus-containing glass microspheres (20-30 µm in diameter) and ferrimagnetic ceramic particles have exhibited great potential to realize minimally invasive treatment of cancer without surgical operation via in situ radiotherapy or hyperthermia of cancer, but it is still a major challenge to clarify the biological reaction between the artificial implants and living body before their application.

Preparation, structure, and in vitro chemical durability of yttrium phosphate microspheres for intra-arterial radiotherapy.

Chemically durable microspheres containing yttrium and/or phosphorus are useful for intra-arterial radiotherapy. In this study, we attempted to prepare yttrium phosphate (YPO4) microspheres with high chemical durability. YPO4 microspheres with smooth surfaces and diameters of around 25 µm were successfully obtained when gelatin droplets containing yttrium and phosphate ions were cooled and solidified in a water-in-oil emulsion and then heat-treated at 1100°C. The chemical durability of the heat-treated microspheres in a simulated body fluid at pH = 6 and 7 was high enough for clinical application of intra-arterial radiotherapy.

Preparation of magnetic iron oxide nanoparticles for hyperthermia of cancer in a FeCl2-NaNO3-NaOH aqueous system.

Magnetic iron oxide nanoparticles (MIONPs) were synthesized in a FeCl2-NaNO3-NaOH aqueous system under various initial Fe(2+)/NO⁻3 molar ratios (α) and Fe(2+)/OH- molar ratios (ß) in order to clarify the effects of the initial molar ratio of reactants on the reaction mechanism. The Fe(2+)/NO⁻3 /OH(-) molar ratio of 3 : 1 : 5 led to the formation of magnetic nanoparticles mainly composed of magnetite (Fe3O4) and maghemite (γ-Fe2O3). The 36 nm sized γ-Fe2O3 and 413 nm sized Fe3O4 were obtained by changing the order in which NaNO3 was added to a NaOH solution. The in vitro heat generations of the resulting MIONPs in an agar phantom were measured under an alternating magnetic field (100 kHz, 23.9 kA/m). The temperature rise (ΔT) of the agar phantom for the 36 nm sized γ-Fe2O3 was 55°C in the first 140 s, with a concentration of 58 mg Fe/mL. Our results showed that it is possible to prepare MIONPs with high heating efficiencies under optimal conditions using the present method.