Papers by Author: William Bonfield

Abstract: Dense and porous HA and Si-HA discs and granules with varying percentages of silicon substitution have been produced and physically and chemically characterised using scanning electron microscopy, surface area analysis, porosimetry, density measurement, image analysis, Xray diffraction, X-ray fluorescence, FT-infrared spectroscopy and in-vitro and in-vivo testing. Results have shown that cell adhesion in-vitro and bone apposition in-vivo are enhanced by the presence of silicon substitution in the hydroxyapatite structure. The biological response to the materials appears to indicate an optimum outcome for levels of silicon substitution of 0.8wt%.

Abstract: Fine nanoapatite relics were deposited on glass substrates by electrohydrodynamic atomisation, using nanohydroxyapatite (nHA), nano-carbonated hydroxyapatite (nCHA) and nanosilicon- substituted hydroxyapatite (nSiHA) suspensions. These electrosprayed nanoapatites were evaluated in-vitro using simulated body fluid (SBF) and human osteoblast (HOB) cells. The SBF study revealed that newly-formed apatite layers were observed on the surface of the relics. Furthermore, enhanced HOB cell growth was observed on each of the nanoapatites at all time points. Hence, this work demonstrated that electrosprayed nanoapatites offer considerable potential as biomaterials.

Abstract: Crystalline hydroxyapatite (HA) and 0.8 wt.% silicon-substituted HA (SiHA) thin films were produced using magnetron co-sputtering. These films were subjected to contact angle measurements and in vitro cell culture study using human osteoblast-like (HOB) cells. A wettability study showed that SiHA has a lower contact angle, and thus is more hydrophilic in nature, as compared to HA. Consequently, enhanced cell growth was observed on SiHA at all time-points. Furthermore, distinct and well-developed actin filaments could be seen within HOB cells on SiHA. Thus, this work demonstrated that the surface properties of the coating may be modified by the substitution of Si into the HA structure.

Abstract: Computerized X-ray microtomography (micro CT) is a powerful technique for imaging and quantification in biomedical materials research. Mapping of various tissue/implants in 3-dimension, which is not achievable with traditional histological slices, is particularly desirable in ex-vivo implant analysis. However, due to a variety of artifacts during image formation, especially the blurred interfaces due to the machine contrast transfer function and the partial volume effect, quantification based on traditional histograms is not accurate. In this work we propose a new 2-dimensional histogram and its application in the analysis of micro CT data of ex-vivo implants, which significantly improve the clarity of the definition of different phases and drastically reduced the artifacts of segmentation compared to conventional methods.

Abstract: Computerized X-ray micro tomography (micro CT) is a powerful technique for studying the structure and properties of porous scaffold. A variety of sample parameters can be studied using a single micro CT scan [1]. In general a segmented data set is a required for most quantitative analysis. However, segmentation of CT data can be difficult due to the artifacts in micro CT images such as blurred interfaces due to the machine contrast transfer function and the partial volume effect. Therefore the segmentation can be biased and prone to errors. Many methods have been developed to improve segmentation, however the interface problem has not been solved perfectly [2]. Porous scaffolds suffer from these effects because of their high surface to volume ratio and hence large interface. In this paper we discuss the interface problem in detail and demonstrate the effect of voxel size on the histograms of CT images of porous scaffold as well as a thresholding method based on 2 dimensional histogram is also presented. The potential of this method in more complicated scenarios such as 3-phase system is currently being investigated [3].

Abstract: A Bioglass® reinforced polyethylene (Bioglass®/polyethylene) composite has been prepared, which combines the high bioactivity of Bioglass® and the toughness of polyethylene. The spatial distribution of Bioglass® particles within the composite is important for the performance of composites in-vivo. Recent developments in X-ray microtomography (XμT) have made it possible to visualize internal and microstructural details with different X-ray absorbencies, nondestructively, and to acquire 3D information at high spatial resolution. In this study, the volume fraction and 3D spatial distribution of Bioglass® particles has been acquired quantitatively by XμT. The information obtained provides a foundation for understanding the mechanical and bioactive properties of the Bioglass®/polyethylene composites.

Abstract: Nano-sized HA (nHA) was applied to the surface of glass and titanium substrates using electrostatic atomisation spray (EAS) deposition. The phase purity of nHA was confirmed by X-ray diffraction. The nHA suspension consisted of rod–like particles 20-30nm in width and 50-100nm in length. The viscosity and conductivity of nHA suspension were 321 mPa s and 5.6 x 10-4 S/m, respectively. EAS of nHA in cone-jet mode was achieved at flow rate of 10-9 m3s-1 with the applied voltage between the needle and the ring-shaped ground electrode of ~6 kV. Micrometer- to submicrometerscaled nHA islands were successfully deposited on the substrate surface. Image analysis showed that the area percentage of nHA increased with deposition time, it covered 50% of the surface area after 10s of spraying. Partial dissolution of nHA was observed after immersion in deionised water for 1 month, particularly on the submicrometer sized nHA islands. Formation of a bone-like apatite layer was found after incubation in simulated body fluid (SBF K9) for 5 days, indicating the high bioactivity of the nHA deposits. In vitro culture with human osteoblast cells showed that the nHA islands were able to support the growth of HOB cells during 7 days of culture; the HOB cell activity increased with culture time as well as EAS deposition time. Immunofluorescence study showed that HOB cells expressed well-organised actin stress fibres on nHA deposited surfaces after 3 days of culture. The result indicated that nHA deposition provided more favourable surfaces for cell attachment. Therefore, electrostatic atomization spray deposition of nHA offers great potential for the creation of bioactive surfaces on bioinert implant surface to provide improved interfacial bonding with host tissues.

Abstract: Nanostructured hydroxyapatite (nHA) thin coatings of thickness 0.5 µm have been successfully produced using a radio-frequency magnetron sputtering technique, through careful selection and control of the processing conditions. nHA coatings were immersed in simulated body fluid (SBF) to determine the rate of nucleation and growth of an apatite layer on their surface. A dense, newlyformed apatite layer with similar characteristics to that of the biological bone apatite, was observed after 7 days of immersion in SBF. X-ray diffraction and infrared analyses confirmed this layer to be calcium-deficient nanocrystalline carbonate HA. All these results demonstrated that the novel nHA coatings were highly bioactive, and the time-frame required to form a dense apatite layer was reduced significantly as compared to the micrometer-sized, sintered HA pellets (from 28 days to 7 days).

Abstract: Silicon-containing apatite (Si-HAp) fibres were successfully synthesized by a homogeneous precipitation method. The resulting Si-HAp fibres were composed of carbonate-containing apatite fibres with preferred orientation in the c-axis. The Si contents in the Si-HAp fibres could be controlled by the Si concentration of the starting solutions. TEM observation indicated that the Si-HAp fibres were of single crystal. The Si-HAp fibres have potential as novel materials for high-performance biomedical devices.

Abstract: The mechanism by which carbonate substitution within the hydroxyapatite (HA) lattice improves osteoconduction is unclear. Discs of dense, sintered, phase-pure HA and carbonate substituted hydroxyapatite (CHA) were cultured with human CD14+ cells in the presence of macrophage-colony stimulating factor (M-CSF) and soluble receptor activator of nuclear factor (NF)-κB (sRANKL), during which time osteoclasts developed and resorbed the ceramic surface. Discs were then seeded with human osteoblasts (HOBs), and proliferation and collagen synthesis measured. Proliferation was increased on resorbed compared to control (unresorbed) surfaces on both materials. Collagen synthesis was increased on CHA compared to HA, an increase accelerated on a previously resorbed surface. The results suggest that osteoclasts can condition synthetic bioceramic surfaces and alter the responses of osteoblasts which subsequently populate them. Carbonate substitution may enhance osteoconduction via effects on enhanced bioresorption.