Surface functionalization of hydroxyapatite nanoparticles for biomedical applications.

This review completely covers the various aspects of hydroxyapatite (HAp) nanoparticles and their role in different biological situations, and provides the surface and interface contents on (i) hydroxyapatite nanoparticles and their hybridization with organic molecules, (ii) surface designing of hydroxyapatite nanoparticles to provide their biocompatibility and photofunction, and (iii) coating technology of hydroxyapatite nanoparticles. In particular, we summarized how the HAp nanoparticles interact with the different ions and molecules and highlighted the potential for hybridization between HAp nanoparticles and organic molecules, which is driven by the interactions of the HAp nanoparticle surface ions with several functional groups of biological molecules. In addition, we highlighted the studies focusing on the interfacial interactions between the HAp nanoparticles and proteins for exploring the enhanced biocompatibility. Such studies focus on how these interactions affect the hydration layers and protein adsorption. However, the hydration layer state involves diverse molecular interactions that can alter the shape of the adsorbed proteins, thereby affecting cell adhesion and spreading on the surfaces. We also summarized the relationship between the surface properties of the HAp nanoparticles and the hydration layer. Furthermore, we spotlighted the cytocompatible photoluminescent probes that can be developed by designing HAp/organic nanohybrid structures. We then emphasized the importance of photofunctionalization in theranostics, which involves the integration of diagnostics and therapy based on the surface design of the HAp nanoparticles. Furthermore, the coating techniques using HAp nanoparticles and HAp nanoparticle/polymer composites were outlined for fusing base biomaterials with biological tissues. The advantages of HAp/biocompatible polymer composite coatings include the ability to effectively cover porous or irregularly shaped surfaces while controlling the thickness of the coating layer, and the addition of HAp nanoparticles to the polymer matrix improves the mechanical properties, increases the roughness, and forms the morphologies that mimic bone nanostructures. Therefore, the fundamental design of hydroxyapatite nanoparticles and their surfaces was suggested from various aspects for biomedical applications.

Mechanochemical Solid-State Immobilization of Photofunctional Dyes on Amorphous Silica Particles and Investigation of Their Interactive Mechanisms.

Amorphous silica particles (ASPs) have been reported to exhibit bioactive properties and are becoming the focus of attention as bioceramics. However, their interactions with proteins in living organisms remain to be understood and need to be investigated in order to achieve wider applications. Our research group found that chlorine (Cl)-containing ASPs are useful for protein immobilization. Photofunctional dyes (fluorescein (FS-), methylene blue (MB+)) that have the carboxy and amino groups as the main functional groups were immobilized on the Cl-containing ASPs via the mechanochemical method as the model molecule and their spectral properties were used to investigate and discuss the organic/inorganic interfacial bonding states. In FS-, the oxygen atoms of the carboxy groups in the molecule were immobilized by the hydrogen bonds with the silanol groups on the ASPs surfaces, indicating that there is an optimum Cl content for the immobilization as the monomer state. In the case of MB+, as the Cl concentration in the ASPs increases, the immobilization via the electrostatic interactions between the Cl in the ASPs and the terminal dimethylamino group, and the hydrogen bonding between the N atoms of the MB+ hetero ring and the particle silanol group were enhanced. These results mainly suggest that the protein adsorption system occurs through the hydrogen bonding between the carboxy groups of the protein and the silanol groups on the particles and via electrostatic interactions between the amino groups of the protein and the dissociated silanol groups and the contained Cl at the particles. Thus, the spectral characterization using dyes as probes is expected to predict the protein interactions with the amorphous silica particles.

Investigation into the Photochemical Properties of Methylene Blue-Immobilized Hydroxyapatite Nanoparticles for Theranostic Application.

In the biomedical field, there has been a requirement for developing theranostic nanomaterials with higher biosafety, leading to both diagnosis and therapy. Methylene blue (MB+) is an organic dye with both photoluminescence (PL) and photosensitization abilities to generate singlet oxygen (1O2). However, MB+ easily loses its generation ability by hydrogen reduction in vivo or by forming aggregates. In this study, MB+ immobilized on biocompatible hydroxyapatite (HA) nanoparticles was applied for the bifunctions of efficient PL and photosensitization. The MB+-immobilized HA nanoparticles (MH) formed aggregates with sizes of 80-100 nm in phosphate buffer (PB). The generation amount and efficiency of 1O2 from the nanoparticles in PB seem to depend on the immobilized MB+ amount and the percentage of the monomer, respectively. Considering the larger immobilized amount and percentage of the MB+ monomer, it was found that there was MH with the lower generation amount and efficiency of 1O2 to exhibit the highest PL intensity. The photofunctional measurement of MB+ revealed the state of MB+ molecules on the HA surface, and it was suggested that the MB+ molecules immobilized on the MH surface would form more hydrogen bonds to change their excitation states. In the cellular experiments, the Hela cancer cells reacted with the nanoparticles and showed red-color PL, indicating cellular imaging. Furthermore, the adherent cell coverage decreased by 1O2 generation, indicating the importance of the immobilization amount of the MB+ monomer. Therefore, theranostic nanomaterials with biosafety were successfully synthesized to show two photofunctions, which provide both cellular imaging and photodynamic therapy by the nanohybrid system between HA and MB+.

Drug Molecular Immobilization and Photofunctionalization of Calcium Phosphates for Exploring Theranostic Functions.

Theranostics (bifunction of therapeutics and diagnostics) has attracted increasing attention due to its efficiency that can reduce the physical and financial burden on patients. One of the promising materials for theranostics is calcium phosphate (CP) and it is biocompatible and can be functionalized not only with drug molecules but also with rare earth ions to show photoluminescence that is necessary for the diagnostic purpose. Such the CP-based hybrids are formed in vivo by interacting between functional groups of organic molecules and inorganic ions. It is of great importance to elucidate the interaction of CP with the photofunctional species and the drug molecules to clarify the relationship between the existing state and function. Well-designed photofunctional CPs will contribute to biomedical fields as highly-functional ormultifunctional theranostic materials at the nanoscales. In this review, we describe the hybridization between CPs and heterogeneous species, mainly focusing on europium(III) ion and methylene blue molecule as the representative photofunctional species for theranostics applications.

Coordination effect of citric acid to Ca-deficient hydroxyapatite on phase transition.

The phase transition of Ca-deficient hydroxyapatite (CDHA) with citric acid (Cit) coordination was investigated. Cit promoted the substitution of K+ ions into CDHA to generate the HA phase. The K+-doping increased the phase transition temperature of CDHA, providing the transition to ß- and α-tricalcium phosphates at higher temperatures. These results suggest controllable phase transition via Cit addition.

Effective Immobilization of Monomeric Methylene Blue on Hydroxyapatite Nanoparticles by Controlling Inorganic-Organic Interfacial Interactions.

We successfully synthesized methylene blue (MB+)-immobilized hydroxyapatite (HM) nanoparticles by changing the initial P/Ca molar ratio. The immobilized amount of MB+ increased with increasing the initial P/Ca molar ratio from 0.6 to 4.0, and the HM had an elliptical shape (long length, 21-24 nm; short length, 11-13 nm) irrespective of the initial P/Ca molar ratio. Upon increasing the initial P/Ca molar ratio, the number of carbonate ions on the HM surface decreased, which would be owing to the electrostatic repulsion by the surface phosphate ions (i.e., P-O-), the surface P-OH mainly dissociated to form P-O-, and the electrostatic interaction of P-O- with MB+ enhanced. The bonding of MB+ with surface P-OH and Ca2+ sites of hydroxyapatite would be hydrogen-bonding and Lewis acid-base interactions, respectively. The optimum synthesis condition for MB+ immobilization at the monomer state was found to be the initial P/Ca molar ratio of 2.0. Here, the existence percentage of the MB+ monomer and the molecular occupancy of the surface carbonate ion at the initial P/Ca molar ratio of 2.0 were higher than those at 4.0 with no significant difference in the immobilized amount of MB+, indicating that MB+ at the initial P/Ca molar ratio of 4.0 is more aggregated than that at 2.0. These results suggested that a part of carbonate ions has a role as a spacer to suppress MB+ aggregation. Accordingly, the interfacial interactions between the MB+ monomer and the hydroxyapatite surface were clarified, which can effectively be controlled by the initial P/Ca molar ratio. These findings will provide fundamental and useful knowledge for the design of calcium phosphate-organic nanohybrids. We believe that these particles will be the base materials to realize diagnostic and/or therapeutic functions through the molecular state control by optimizing the synthesis conditions.

Preparation of Monodispersed Nanoporous Eu(III)/Titania Loaded with Ibuprofen: Optimum Loading, Luminescence, and Sustained Release.

Functional nanomaterials are one of the potential carriers for drug delivery, whereas there are many prerequisites for this purpose. The carrier should be monodispersed, be fluorescent, and have a proper nanostructure to keep/release drug molecules to achieve controlled release, although preparing a nanomaterial which fulfills all the demands is still very challenging. In this paper, we show the preparation of monodispersed nanoporous amorphous titania submicron particles with fluorescent property. They adsorb a model drug molecule-ibuprofen-with their surface coverage up to 100%. Such a perfect loading does not decrease the fluorescent intensity because of any quenching effects but even maximize it. We also demonstrate the release behavior of IBU into simulated body fluid. Interestingly, the present carrier releases most of IBU in 6 h, whereas that modified with the polyethylene glycol moiety takes 48 h to finish releasing IBU, indicating its potential for controlled release applications.