HAp Nanoparticles Research Articles

Fabrication and Characterization of Magnetic Hydroxyapatite Entrapped Agarose Composite Beads with High Adsorption Capacity for Heavy Metal Removal

Magnetic hydroxyapatite entrapped agarose composite beads (M-HAP/Agar composite beads) have been successfully synthesized by emulsification of magnetic HAP nanoparticles with agarose suspension. In the process, the magnetic HAP nanoparticles served as the main resource for high adsorption performance, which were constructed by surface modification of Fe3O4 with N-(phosphonomethyl)iminodiacetic acid (PM-IDA) and followed by coating with HAP. This strategy integrates the distinct advantages of large-size beads and magnetic response for easy recovery as well as nanoparticles for high adsorption capacity. The resulting M-HAP/Agar composite beads display large specific surface area (90 m2 g–1) and uniform spherical shape (150 μm). These magnetic hydroxyapatite nanoparticles in beads can provide more adsorption sites due to their suitable porous structure. As a result, the adsorbent exhibits excellent performance in adsorption of Pb2+, Co2+, and Cu2+, showing maximum binding capacities as high as 842.6, 105.1, ...

Rapid synthesis of citrate-zinc substituted hydroxyapatite using the ultrasonication-microwave method

Abstract Bio-inspired citrate-hydroxyapatite (HAP) nanoparticles doped with zinc ions were prepared from mussel shells via a rapid method of ultrasonication and microwave irradiation. It was found that zinc ions can interact with citrate ions in the crystallization process of HAP. The growth of nanocrystals in citrate solution was slower than that in HCl solution. The addition of zinc ions reduced the residue of citrate in the products and increased the crystallinity of HAP. Furthermore, mineral platelets with the length of 29 ± 7 nm and the thickness of 3–5 nm were obtained for Zn/(Zn+Ca) = 5 mol%, which displayed similar morphology with HAP platelets in bone. Moreover, the obtained HAP nanoparticles are promising for use as bone graft materials in biomedical applications.

Artificial enamel induced by phase transformation of amorphous nanoparticles

Human tooth enamel has tightly packed c-axis-oriented hydroxyapatite (HAP: Ca10(PO4)6(OH)2) nanorods with high elastic modulus. Fabrication of an enamel architecture in vitro supports the repair of teeth using HAP; however, existing methods require complex and laborious steps to form an enamel-like structure. Here we present a very simple and effective technique for forming artificial enamel in near-physiological solution using a substrate composed of amorphous calcium phosphate (ACP) nanoparticles. Without any functionalized modification of the substrate surface, faint dissolution and successive phase transformation automatically induce formation of an intermediate layer of low-crystalline HAP nanoparticles, on which highly oriented HAP nanorods grow by geometrical selection. We also show that an enamel structure forms on a substrate of amorphous calcium carbonate when the surface nanoparticles react so as to form an intermediate layer similar to that in ACP. Our results demonstrate that there is a wide range of substrate choices for nanorod array formation. Contrary to current understanding, a stable surface designed in nanoscale is not essential for the growth of arranged guest crystals. Reactive amorphous nanoparticles and their transformation efficiently induce a nanorod array structure.

The influence of the preparation conditions and filler content on thermal properties of poly-l -lactide and hydroxyapatite/poly-l -lactide nanocomposite

Poly-l-lactide (PLLA) and hydroxyapatite/poly-l-lactide (HAp/PLLA) are two widely used biomaterials for three-dimensional scaffolds, drug release matrices and implantable medical devices for reparation of bone tissue; diversity in the initial preparation and filler content has a significant influence on different properties such as morphology and crystallinity, thus playing a considerable role in most of these applications. For this reason, PLLA and HAp/PLLA samples with a large difference in crystallinity (from below 20% to over 70%) and filler content (up to 86 wt% of HAp nanoparticles with an average diameter of 80 nm) were prepared and consequent dissimilarities in morphology, crystallinity and thermal properties were investigated by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), wide angle X-ray diffraction (WAXD) measurements and Fourier transform infrared (FTIR) spectroscopy. Special attention was devoted to analyzing data obtained from thermal measurements. A three-phase model was employed in order to describe the heat capacity step decline in the nanocomposite; the evolution in different polymer fractions, the crystalline fraction and the mobile and rigid amorphous fractions, with filler content was determined. © 2017 Society of Chemical Industry

The Influence of Nanohydroxyapatite on the Thermal, Mechanical, and Tribological Properties of Polyoxymethylene Nanocomposites

The influence of nanohydroxyapatite on the glass transition region and its activation energy, as well as on the tribological and mechanical properties of polyoxymethylene nanocomposites, was investigated using DMA, TOPEM DSC, nanoindentation, and nondestructive ultrasonic methods. It was found that the glass transition for unmodified POM was in the lower temperature range than in POM/HAp nanocomposites. Moreover,ΔCpand activation energy were larger for POM/HAp nanocomposites. Friction coefficient was higher for POM/HAp nanocomposites in comparison to both POM homopolymer and POM copolymer. Simultaneously, the indentation test results show that microhardness is also higher for POM/HAp nanocomposites than for POM. From ultrasonic investigations it was found that the highest values of both longitudinal and transverse propagation waves and Young’s and shear modulus for POM homopolymer (DH) and POM copolymer T2H and their nanocomposites can be attributed to their higher degree of crystallinity in comparison to UH copolymer. Moreover, for POM/HAp nanocomposites with 5% of HAp, ultrasonic longitudinal wave velocity was almost constant even after 1000000 mechanical loading cycles, evidencing an enhancement of mechanical properties by HAp nanoparticles.

Newly Developed Biocompatible Material: Dispersible Titanium-Doped Hydroxyapatite Nanoparticles Suitable for Antibacterial Coating on Intravascular Catheters.

Thirteen patients with chlorhexidine-silver sulfadiazine-impregnated catheters have experienced serious anaphylactic shock in Japan. These adverse reactions highlight the lack of commercially available catheters impregnated with strong antibacterial chemical agents. A system should be developed that can control both biocompatibility and antibacterial activity. Hydroxyapatite (HAp) is biocompatible with bone and skin tissues. To provide antibacterial activity by using an external physical stimulus, titanium (Ti) ions were doped into the HAp structure. Highly dispersible, Ti-doped HAp (Ti-HAp) nanoparticles suitable as a coating material were developed. In 3 kinds of Ti-HAp [Ti/(Ca + Ti) = 0.05, 0.1, 0.2], the Ti content in the HAp was approximately 70% of that used in the Ti-HAp preparation, as determined by inductively coupled plasma atomic emission spectroscopy (ICP-AES). ICP-AES and X-ray diffraction showed Ti ions were well substituted into the HAp lattice. The nanoparticles were almost uniformly coated on a polyethylene (PE) sheet in a near-monolayer with a surface coverage ratio >95%. The antibacterial activity of the Ti-HAp nanoparticles containing 7.3% Ti ions and coating the sheet was evaluated by calculating the survival ratio of Pseudomonas aeruginosa on the coated sheet after ultraviolet (UV) irradiation. The Ti-HAp-coated sheet showed a 50% decrease in the number of P. aeruginosa compared with that on an uncoated control PE sheet after UV irradiation for 30 s. Key Messages: A system of biocompatibility and antibacterial activity with an on/off switch controlled by external UV stimulation was developed. The system is expected to be applicable in long-term implanted intravascular catheters.

Synthesis of nano-hydroxyapatite sorbent for microextraction in packed syringe of phthalate esters in water samples

An easy alkoxide–based sol–gel method using Ca(NO3)2·4H2O and tri–ethyl phosphate (PO(OC2H5)3; TEP) precursors employed to synthesize nano-hydroxyapatite (HAP). The structural characterization of samples was studied using XRD, TEM and elemental analysis of SEM. Scherrer and Williamson–Hall equations were applied to measurement of the crystallite size distributions and micro–strain of the synthesized HAP powder. The synthesized HAP nanoparticles were applied as an efficient sorbent for the microextraction in packed syringe (MEPS) of phthalate esters (PAEs) from water samples. Separation and determination of the PAEs were performed by gas chromatography–flame ionization detection (GC–FID). Several variables affecting the extraction efficiency of the analytes i.e., desorption solvent, volume of desorption solvent and number of extraction cycles were investigated and optimized. Under optimized conditions, the calibration curves were linear (R > 0.9916) in the concentration range of 0.05–100 ng mL−1. The method displays detection limits (at an S/N ration of 3) in the range from 0.02 to 0.1 ng mL−1, and the limits of quantification (at an S/N ratio of 10) are between 0.07 and 0.25 ng mL−1. Relative standard deviations (RSDs) for intra- and inter-day precision are 4.8–8.3%, and 6.1–8.5%, respectively. The applicability of the developed method was examined by analyzing different water samples (river water, bottled mineral water and tap water) and the relative recovery values for the spiked water samples were found to be in the range of 85.5–99.2%.

Synthesis, Characterization, and Atenolol Delivery Application of Functionalized Mesoporous Hydroxyapatite Nanoparticles Prepared by Microwave-Assisted Co-precipitation Method.

Atenolol has been used to treat angina and hypertension, either alone or with other antihypertensives. Despite its usefulness, it shows some side effects such as diarrhea and nausea in some patients. A method for slow release of atenolol in intestine is helpful to prevent such side effects. A facile co-precipitation microwave-assisted method was used to fabricate mesoporous hydroxyapatite nanoparticles (mHAp). It was then functionalized to have SO3H groups. The synthesized material was used for storage/slow release study of atenolol. Atenolol loaded mHAp shows immediate release of atenolol in pH 8, whileafter functionalizing shows up to ca. 30% release at the beginning. In pH 1, 50% of drug was released after 10 h from AT@mHAp and after 18h the drug was almost completely released.The drug release profiles of functionalized HAp at pH value 1 and 8reveals the complete release of atenolol in intestine pH, while no complete release is observed in stomach environment. The aims of this work were synthesis and characterization of mesoporous HAp through the microwave-assisted co-precipitation method and elucidate the underlying drug release capability of mesoporous HAp nanoparticles. The SO3H group was incorporated into the mesoporous HAp and then used as drug delivery carriers using atenolol as a model drug to investigate their drug storage/release properties in simulated body fluid (SBF). Increasing pH value to 8 causes increase in the drug release.

Interaction between functional nano-hydroxyapatite and cells and the underlying mechanisms

To explore the interaction between arginine functionalized hydroxyapatite (HAP/Arg) nanoparticles and endothelial cells, and to investigate mechanisms for endocytosis kinetics and endocytosis. Human umbilical vein endothelial cells (HUVECs) were selected as the research model.Cellular uptake of HAP/Arg nanoparticles were observed by laser scanning confocal microscopy.Average fluorescence intensity of cells after ingestion with different concentrations of HAP/Arg nanoparticles were determined by flow cytometer and atomic force microscopy. The HAP/Arg nanoparticles with doped terbium existed in cytoplasm, and most of them distributed around the nucleus area after cellular uptake by HUVECs. Cellular uptake process of HAP/Arg nanoparticles in HUVECs was in a time and concentration dependent manner. 4 h and 50 mg/L was the best condition for uptake. HAP/Arg nanoparticles were easier to be up-taken into the cells than HAP nanoparticles without arginine functionalized. HAP/Arg nanoparticles are internalized by HUVECs cells through an active transport and energy-dependent endocytosis process, and it is up-taken by cells mainly through caveolin-mediated endocytosis, but the clathrin-dependent endocytic pathway is also involved..

Hydroxyapatite particles as carriers for 223Ra

Systematic investigation of optimal conditions for preparation and in vitro stability of HAP particles labeled with 223Ra, that could be considered as promising candidates for targeted α-therapy, has been carried out. Two different approaches to HAP labelling were tested: sorption of Ra2+ on pre-synthesized HAP-particles and incorporation of Ra2+ into the structure of HAP during its synthesis. Two textural forms of HAP particles were used—nanoparticles and particles with the diameter of 350 ± 20 μm. Kinetics of 223Ra sorption on HAP of different particle size and desorption in 0.9 % NaCl solution were studied. The influence of solution acidity and solid to liquid phase ratio on sorption of Ra was evaluated and the sorption yield up to 98 % was achieved. It was found that the optimal conditions for the sorption included synthesis of HAP nanoparticles in the presence of 223Ra at pH values of 4–7 followed by annealing at 900 °C. In this case subsequent cumulative desorption of Ra was <5 % of initial activity.

Synthesis and characterization of hydroxyapatite nanoparticles templated by ozonolysed natural rubber latex

Nanoscale hydroxyapatites (HAps) were successfully synthesized by means of sol–gel method in the presence of ozonolysed natural rubber latex templates of various molecular weights. The thermal properties, functionality, crystallinity and morphology of HAp powder were characterized by simultaneous thermogravimetry and differential scanning calorimetry, Fourier transform infrared spectroscopy, X-ray diffraction and transmission electron microscopy. The results revealed that lower molecular weight ozonolysed natural rubber templates produced spheroidal and less crystalline HAp nanoparticles. A higher molecular weight template produced HAp nano-rod-like particles with higher crystallinity. The results obtained in this work suggest that the molecular weight as well as functionality of the biomacromolecule template affect the phase crystallinity and morphology of synthesized HAp.

Influence of concentration of hydroxyapatite surface modifier agent on bioactive composite characteristics

The characterization of chitosan – hydroxyapatite (CH – HAp) composite sponges prepared via freeze-drying methodology is reported in this study. Stearic acid (SA), added as a surface modifier of the HAp nanoparticles, induced changes in the TG/DTG results, particle size distribution and particle morphology. Composite sponges prepared with SA coated HAp demonstrated enhanced biocompatibility and structural properties, as compared to the composites prepared with uncoated HAp. SA coating modified the morphology of the composite, promoting a better dispersion of HAp particles within the composite sponges, and better homogeneity of the polymeric cover with HAp particles. The viability of the composites for cell culture applications was analyzed, and the results suggest that the sponges are biocompatible. Therefore, SA proved to be a good candidate for surface coating of HAp nanoparticles prevent agglomerations, and could be used effectively in the preparation of biocompatible composite sponges with chitosan.

Engineered electrospun poly(caprolactone)/polycaprolactone-g-hydroxyapatite nano-fibrous scaffold promotes human fibroblasts adhesion and proliferation

Polycaprolactone (PCL)/hydroxyapatite nano-composites are among the best candidates for tissue engineering. However, interactions between nHAp and PCL are difficult to control leading to inhomogeneous dispersion of the bio-ceramic particles. Grafting of polymer chains at high density/chain length while promotes the phase compatibility may result in reduced HAp exposed surface area and therefore, bioactivity is compromised. This issue is addressed here by grafting PCL chains onto HAp nano-particles through ring opening polymerization of ε-caprolactone (PCL-g-HAp). FTIR and TGA analysis showed that PCL (6.9wt%), was successfully grafted on the HAp. PCL/PCL-g-HAp nano-fibrous scaffold showed up to 10 and 33% enhancement in tensile strength and modulus, respectively, compared to those of PCL/HAp. The effects of HAp on the in vitro HAp formation were investigated for both the PCL/HAp and PCL/PCL-g-HAp scaffolds. Precipitation of HAp on the nano-composite scaffolds observed after 15days incubation in simulated body fluid (SBF), as confirmed by scanning electron microscopy (SEM), and energy dispersive X-ray spectroscopy (EDX).Human fibroblasts were seeded on PCL, PCL/HAp and PCL/PCL-g-HAp scaffolds. According to MTT assay, the highest cell proliferation was recorded for PCL/PCL-g-HAp nano-composite, at all time intervals (1–21days, P<0.001). Fluorescent microscopy (of DAPI stained samples) and electron microscopy images showed that all nano-fibrous scaffolds (PCL, PCL/HAp, and PCL/PCL-g-HAp), were non-toxic against cells, while more cell adhesion, and the most uniform cell distribution observed on the PCL/PCL-g-HAp.Overall, grafting of relatively short chains of PCL on the surface of HAp nano-particles stimulates fibroblasts adhesion and proliferation on the PCL/PCL-g-HAp nano-composite.

Hafnium-doped hydroxyapatite nanoparticles with ionizing radiation for lung cancer treatment

Recently, photodynamic therapy (PDT) is one of the new clinical options by generating cytotoxic reactive oxygen species (ROS) to kill cancer cells. However, the optical approach of PDT is limited by tissue penetration depth of visible light. In this study, we propose that a ROS-enhanced nanoparticle, hafnium-doped hydroxyapatite (Hf:HAp), which is a material to yield large quantities of ROS inside the cells when the nanoparticles are bombarded with high penetrating power of ionizing radiation. Hf:HAp nanoparticles are generated by wet chemical precipitation with total doping concentration of 15mol% Hf4+ relative to Ca2+ in HAp host material. The results show that the HAp particles could be successfully doped with Hf ions, resulted in the formation of nano-sized rod-like shape and with pH-dependent solubility. The impact of ionizing radiation on Hf:HAp nanoparticles is assessed by using in-vitro and in-vivo model using A549 cell line. The 2′,7′-dichlorofluorescein diacetate (DCFH-DA) results reveal that after being exposed to gamma rays, Hf:HAp could significantly lead to the formation of ROS in cells. Both cell viability (WST-1) and cytotoxicity (LDH) assay show the consistent results that A549 lung cancer cell lines are damaged with changes in the cells’ ROS level. The in-vivo studies further demonstrate that the tumor growth is inhibited owing to the cells apoptosis when Hf:HAp nanoparticles are bombarded with ionizing radiation. This finding offer a new therapeutic method of interacting with ionizing radiation and demonstrate the potential of Hf:HAp nanoparticles in tumor treatment, such as being used in a palliative treatment after lung surgical procedure. Statement of SignificancePhotodynamic therapy (PDT) is one of the new clinical options by generating cytotoxic reactive oxygen species (ROS) to kill cancer cells. Unfortunately, the approach of PDT is usually limited to the treatment of systemic disease and deeper tumor, due to the limited tissue penetration depth of visible light (620–690nm). Here we report a ROS-enhanced nanoparticle, hafnium-doped hydroxyapatite (Hf:HAp), which can trigger ROS when particles are irradiated with high penetrating power of ionizing radiation. The present study provides quantitative data relating ROS generation and the therapeutic effect of Hf:HAp nanoparticles in lung cancer cells. As such, this material has opened an innovative window for deeper tumor and systemic disease treatment.

Degenerative Scoliosis de Novo: Prospective Study of 50 Cases Operated with Stand-Alone ALIF with One Years Minimum Follow-Up

Introduction Posterior approach in degenerative scoliosis de novo is related to high rate of complications in elderly people. In a previous retrospective and comparative report in 2009 (French Spinal Society), Stand alone ALIF/OLIF proved a less iatrogenic procedure to relief the pain and meet patient's expectations. A prospective study including 50 patients has been performed to confirm this previous report, with at least one-year follow-up. Material and Methods The indications are based on a specific classification according to the biomechanism of the rotatory discogenic collapse. Therefore, two homogenic groups are compared. The details of the procedure are important with elderly people to avoid vascular complication or subsidence (preventive vertebral augmentation) due to osteoporosis. Stand alone ALIF/OLIF using a Peek cage. The cage and disk space are filled with nanoparticles of HAP. The outcomes are assessed with VAS, ODI, Kim&amp;Kim score, standard radiography and spinogram at two, six and twelve-month follow-up periods. Results The two groups have specific physiopathology and epidemiology. But, outcomes are very similar after one-year follow-up. Surprisingly, it is not related to the number of disk's levels (1 to 4) operated on. According to Kim&amp;Kim score: 30% have excellent outcome, 60% good result. VAS drops from 7/10 to 2/10 at one year. Complications (10% poor results) are mainly related to vertebral body fractures (upper endplate of L5). Posterior Vertebral augmentation was not efficient to improve the score. Symptomatic effect of sympathectomy is noted (8%) but with good acceptance. Revision rate is 6% mainly due to the persistence of a radiculopathy linked to posterior stenosis. A posterior decompression was performed after 4 months and relieved the symptoms. Radiologic fusion rate is 90% with HAP nanoparticles. It proves a low cost, less iatrogenic than BMP 2 and efficient solution in elderly people. Conclusion This prospective study confirms that Stand alone ALIF/OLIF is a better procedure to relieve the pain and meet patient's expectation compared with posterior instrumented surgery. Preserving posterior musculature and reshaping the anterior “lumbar arch,” Stand alone ALIF improves the sagittal balance in a physiological way. Osteoporosis required a “systematic” vertebral augmentation by anterior approach to lower subsidence or vertebral body fractures rate.

Spray drying as a viable process to produce nano-hydroxyapatite/chitosan (n-HAp/CS) hybrid microparticles mimicking bone composition

In this work nano-hydroxyapatite/chitosan (n-HAp/CS) hybrid microparticles were prepared by spray drying following a methodology where, in a first step, aqueous nanodispersions of n-HAp in the presence of chitosan were produced by fast stirring at pH values below and above chitosan pKa (5.5 and 7.0, respectively). The mixing system used allowed the production of homogeneous and stable nanodispersions, and the subsequent spray-dried microparticles, incorporating highly pure HAp nanoparticles of approximately 50nm, were successfully produced without degrading the organic component, chitosan. Comparatively with the n-HAp/CS-7.0, the n-HAp/CS-5.5 dispersions were characterized by a lower particle size and a higher zeta potential, being then more stable. Differences in the spray-dried microparticles were also evident from a morphological point of view. HApCS-5.5 microparticles, which present an average size in volume of 15.8μm and n-HAp crystals homogenously distributed, were found to be preferred over the HApCS-7.0 counterparts, which require an extra step in the productive process and presented a tendency to form large agglomerates. Both prepared hybrid particles presented similar composition to that one of natural bone (HAp/CS of 70/30) and no traces of KCl salts were observed if a washed n-HAp paste was used.

HAp incorporated ultrafine polymeric fibers with shape memory effect for potential use in bone screw hole healing.

In the clinical setting of bone fracture healing, hardware removal often causes localized microtrauma and residual screw holes may act as stress risers to place the patient at a risk of refracture. To address this noted issue, this study proposed to develop a biologically mimicking and mechanically self-actuated nanofibrous screw-like scaffold/implant for potential in situ bone regeneration. By incorporating nano-hydroxyapatite (HAp) into a shape memory copolymer poly(d,l-lactide-co-trimethylene carbonate) (PLMC) via co-electrospinning, composite nanofibers of HAp/PLMC with various HAp proportions (1, 2 and 3 wt%) were successfully generated. Morphological, thermal and mechanical properties as well as the shape memory effect of the resultant HAp/PLMC nanofibers were characterized using a variety of techniques. Thereafter, osteoblasts isolated from rat calvarial were cultured on the fibrous HAp/PLMC scaffold to assess its suitability for bone regeneration in vitro. We found that agglomerates gradually appeared on the fiber surface with increasing HAp loading fraction. The switching temperature for actuating shape recovery Ts (i.e., glass transition temperature Tg) of the fibrous HAp/PLMC was readily modulated to fall between 43.5 and 51.3 °C by varying the HAp loadings. Excellent shape memory properties were achieved for the HAp/PLMC composite nanofibers with a shape recovery ratio of Rr > 99% and shape fixity ratio of Rf > 99%, and the shape recovery force of the HAp/PLMC nanofibers was also strengthened compared to that of the HAp-free PLMC nanofibers. Moreover, we demonstrated that the engineered screw-like HAp/PLMC scaffold/implant (ϕ = 5 mm) was able to return from a slender bar to its original stumpy shape in a time frame of merely 8 s at 48 °C. Biological assay results corroborated that the incorporation of HAp to PLMC nanofibers significantly enhanced the alkaline phosphatase secretion as well as mineral deposition in bone formation. These attractive results warrant further investigation in vivo on the feasibility of applying the biomimicking nanofibrous HAp/PLMC scaffold with shape memory effect for bone screw hole healing.

Preparation of chitosan/nano hydroxyapatite organic–inorganic hybrid microspheres for bone repair

In this work, we encapsulated icariin (ICA) into chitosan (CS)/nano hydroxyapatite (nHAP) composite microspheres to form organic–inorganic hybrid microspheres for drug delivery carrier. The composition and morphology of composite microspheres were characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM) and differential scanning calorimetry– thermogravimetric analysis (DSC–TGA). Moreover, we further studied the performance of swelling properties, degradation properties and drug release behavior of the microspheres. ICA, the extract of traditional Chinese medicine-epimedium, was combined to study drug release properties of the microspheres. ICA loaded microspheres take on a sustained release behavior, which can be not only ascribed to electrostatic interaction between reactive negative hydroxyl (OH) of ICA and positive amine groups (NH2) of CS, but also depended on the homogeneous dispersion of HAP nanoparticles inside CS organic matrix. In addition, the adhesion and morphology of osteoblasts were detected by inverted fluorescence microscopy. The biocompatibility of CS/nHAP/ICA microspheres was evaluated by the MTT cytotoxicity assay, Hoechst 33258 and PI fluorescence staining. These studies demonstrate that composite microspheres provide a suitable microenvironment for osteoblast attachment and proliferation. It can be speculated that the ICA loaded CS-based organic–inorganic hybrid microspheres might have potential applications in drug delivery systems.

Gene polA as a Suitable Reference for Studying Antibacterial Effect of Hydroxyapatite.

Hydroxyapatite (HAP: Ca10(PO4)6(OH)2) is a widely used biocompatible material; however, information on the reaction mechanism between HAP and microorganisms is insufficient. This study aimed to identify a stable reference gene for studying the antibacterial activity of HAP. The half maximal inhibitory concentration (C50) and gene expression of a human symbiotic Escherichia coli strain TOP10, was investigated at various concentrations of HAP. Our uniformly sized HAP nanoparticles (HANPs) had a high surface area and an estimated IC50 of 75 mg/mL. The expressions of genes, including those of DNA polymerase I (poIA), DNA polymerase II (poIB), cytochrome d complex (cyd), glucan biosynthesis protein G (mdoG), D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and 16S ribosomal RNA (16S rRNA), were analyzed by performing quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and using reported and newly designed primers. The changes in the copy numbers compared to non-HANPs-treated conditions for polB, cyd, mdoG, GAPDH, and 16S rRNA were 50%-381%, 0.7%-66.3%, 1.1%-7.8%, 1.6%-86.3%, and 0.3%-8.1%, respectively. The expressions of polA remained stable under all conditions (62.8%-88.4%). Therefore, we identified polA as a suitable reference gene. Moreover, the expressions of cyd and mdoG were inhibited, indicating that the antibacterial activity of HANPs is related to cell membrane or cell wall proteins. Our findings provide a thorough understanding of HAP-microorganism interactions for potential biomedical applications.

Effect of the Materials Properties of Hydroxyapatite Nanoparticles on Fibronectin Deposition and Conformation.

Hydroxyapatite (HAP, Ca10(PO4)6(OH)2) nanoparticles with controlled materials properties have been synthesized through a two-step hydrothermal aging method to investigate fibronectin (Fn) adsorption. Two distinct populations of HAP nanoparticles have been generated: HAP1 particles had smaller size, plate-like shape, lower crystallinity, and more negative ζ potential than HAP2 particles. We then developed two-dimensional platforms containing HAP and Fn and analyzed both the amount and the conformation of Fn via Förster resonance energy transfer (FRET) at various HAP concentrations. Our FRET analysis reveals that larger amounts of more compact Fn molecules were adsorbed onto HAP1 than onto HAP2 particles. Additionally, our data show that the amount of compact Fn adsorbed increased with increasing HAP concentration due to the formation of nanoparticle agglomerates. We propose that both the surface chemistry of single nanoparticles and the size and morphology of HAP agglomerates play significant roles in the interaction of Fn with HAP. Collectively, our findings suggest that the HAP-induced conformational changes of Fn, a critical mechanotransducer protein involved in the communication of cells with their environment, will ultimately affect downstream cellular behaviors. These results have important implications for our understanding of organic–inorganic interactions in physiological and pathological biomineralization processes such as HAP-related inflammation.