A Porous Fluoride-Substituted Bovine-Derived Hydroxyapatite Scaffold Constructed for Applications in Bone Tissue Regeneration.

Hydroxyapatite is widely used in bone implantation because of its similar mineral composition to natural bone, allowing it to serve as a biocompatible osteoconductive support. A bovine-derived hydroxyapatite (BHA) scaffold was developed through an array of defatting and deproteinization procedures. The BHA scaffold was substituted with fluoride ions using a modified sol-gel method to produce a bovine-derived fluorapatite (BFA) scaffold. Fourier-transform infrared spectroscopy and X-ray diffraction analysis showed that fluoride ions were successfully substituted into the BHA lattice. According to energy dispersive X-ray analysis, the main inorganic phases contained calcium and phosphorus with a fluoride ratio of ~1-2 wt%. Scanning electron microscopy presented a natural microporous architecture for the BFA scaffold with pore sizes ranging from ~200-600 µm. The BHA scaffold was chemically stable and showed sustained degradation in simulated-body fluid. Young's modulus and yield strength were superior in the BFA scaffold to BHA. In vitro cell culture studies showed that the BFA was biocompatible, supporting the proliferative growth of Saos-2 osteoblast cells and exhibiting osteoinductive features. This unique technique of producing hydroxyapatite from bovine bone with the intent of producing high performance biomedically targeted materials could be used to improve bone repair.

Chitosan: A review of molecular structure, bioactivities and interactions with the human body and micro-organisms.

Chitosan has many desirable attributes e.g. antimicrobial properties and promoting wound healing, and is used in various applications. This article first discusses how degree of deacetylation (DD) and molecular weight (MW) impacts on what level of bioactivities chitosan manifests, then introduces the "molecular chain configuration" model to explain various possible mechanisms of antimicrobial interactions between chitosan with different MW and different types of bacteria. Similarly, the possible pathways of how chitosan reacts with cancer and the body's immune system to demonstrate immune and antitumor effects are also discussed by using this model. Moreover, the possible mechanisms of how chitosan enhances coagulation and wound healing are also discussed. With these beneficial bioactivities in mind, the application of chitosan in surgery, tissue engineering and oncology is outlined. This review concludes that as chitosan demonstrates many beneficial bioactivities via multiple mechanisms, it is an important polymer with a promising future in medicine.

Chitosan: A review of sources and preparation methods.

Chitosan, derived from chitin, has many desirable biomedical attributes. This review aims to explore different sources of chitin and methods of chitosan production with industrial consideration. This article first discussed different sources of chitin for industrial scale production, with considerations given to both their environmental impacts and commercialization potential. Secondly, this article reviews the two categories of chitosan preparation - chemical methods and biological methods - based on existing publications which used lobster by-products as a feedstock source. The mechanisms of the chemical methods are firstly summarized, and then the different chemical agents and reaction parameters used are discussed. Next, both enzymatic and fermentation-based approaches are reviewed under biological methods and compared with chemical methodologies, with lactic fermentation methods as the major focus. This article concludes that lobster cephalothorax could be an ideal source for chitosan preparation on an industrial scale; and chemical methods involve simpler processing overall, while producing chitosan with stronger bioactivities because of the lower molecular weight (MW) and higher degree of deacetylation (DD) achieved by the products. Moreover, due to biological methods inevitably necessitating further chemical processing, an approach involving some unconventional chemical methods has been regarded as a more suitable strategy for industrial scale chitosan production.

Study of biomorphic calcium deficient hydroxyapatite fibres derived from a naturalHarakeke(Phormium tenax) leaf fibre template.

The complex structure of natural bio-organic matter has inspired scientists to utilise these as templates to design 'biomorphic materials', which retain the intricate architecture of the materials while acting as a useful bioactive material. Biomorphic hydroxyapatite-based fibres were synthesised usingHarakekeleaf fibre as a template, which constitutes a powerful method for manufacturing bioactive ceramic fibres. Furthermore, in creating the hydroxyapatite-based fibres, a natural source of calcium and phosphate ions (from bovine bone) was utilised to create the digest solution in which the leaf fibres were immersed prior to their calcination to form the inorganic fibres. Chemical, thermogravimetric and microscopic characterisation confirmed that the final product was able to successfully replicate the shape of the fibres and furthermore be transformed into calcium deficient, bone-like hydroxyapatite.

A Review of The Lesser-Studied Microemulsion-Based Synthesis Methodologies Used for Preparing Nanoparticle Systems of The Noble Metals, Os, Re, Ir and Rh.

This review focuses on the recent advances in the lesser-studied microemulsion synthesis methodologies of the following noble metal colloid systems (i.e., Os, Re, Ir, and Rh) using either a normal or reverse micelle templating system. The aim is to demonstrate the utility and potential of using this microemulsion-based approach to synthesize these noble metal nanoparticle systems. Firstly, some fundamentals and important factors of the microemulsion synthesis methodology are introduced. Afterward, a review of the investigations on the microemulsion syntheses of Os, Re, Ir, and Rh nanoparticle (NP) systems (in all forms, viz., metallic, oxide, mixed-metal, and discrete molecular complexes) is presented for work published in the last ten years. The chosen noble metals are traditionally very reactive in nanosized dimensions and have a strong tendency to aggregate when prepared via other methods. Also, the particle size and particle size distribution of these colloids can have a significant impact on their catalytic performance. It is shown that the microemulsion approach has the capability to better stabilize these metal colloids and can control the size of the synthesized NPs. This generally leads to smaller particles and higher catalytic activity when they are tested in applications.

Special Issue: Novel Advances and Approaches in Biomedical Materials Based on Calcium Phosphates.

Research on calcium phosphate use in the development and clinical application of biomedical materials is a diverse activity and is genuinely interdisciplinary, with much work leading to innovative solutions for improvement of health outcomes. This Special Issue aimed to summarize current advances in this area. The nine papers published cover a wide spectrum of topical areas, such as (1) remineralisation pastes for decalcified teeth, (2) use of statins to enhance bone formation, (3) how dolomitic marble and seashells can be processed into bioceramic materials, (4) relationships between the roughness of calcium phosphate surfaces and surface charge with the effect on human MRC osteogenic differentiation and maturation being investigated, (5) rheological and mechanical properties of a novel injectable bone substitute, (6) improving strength of bone cements by incorporating reinforcing chemically modified fibres, (7) using adipose stem cells to stimulate osteogenesis, osteoinduction, and angiogenesis on calcium phosphates, (8) using glow discharge treatments to remove surface contaminants from biomedical materials to enhance cell attachment and improve bone generation, and (9) a review on how classically brittle hydroxyapatite based scaffolds can be improved by making fibre-hydroxyapatite composites, with detailed analysis of ceramic crack propagation mechanisms and its prevention via fibre incorporation in the hydroxyapatite.

A Review on the Use of Hydroxyapatite-Carbonaceous Structure Composites in Bone Replacement Materials for Strengthening Purposes.

Biomedical materials constitute a vast scientific research field, which is devoted to producing medical devices which aid in enhancing human life. In this field, there is an enormous demand for long-lasting implants and bone substitutes that avoid rejection issues whilst providing favourable bioactivity, osteoconductivity and robust mechanical properties. Hydroxyapatite (HAp)-based biomaterials possess a close chemical resemblance to the mineral phase of bone, which give rise to their excellent biocompatibility, so allowing for them to serve the purpose of a bone-substituting and osteoconductive scaffold. The biodegradability of HAp is low (Ksp ≈ 6.62 × 10-126) as compared to other calcium phosphates materials, however they are known for their ability to develop bone-like apatite coatings on their surface for enhanced bone bonding. Despite its favourable bone regeneration properties, restrictions on the use of pure HAp ceramics in high load-bearing applications exist due to its inherently low mechanical properties (including low strength and fracture toughness, and poor wear resistance). Recent innovations in the field of bio-composites and nanoscience have reignited the investigation of utilising different carbonaceous materials for enhancing the mechanical properties of composites, including HAp-based bio-composites. Researchers have preferred carbonaceous materials with hydroxyapatite due to their inherent biocompatibility and good structural properties. It has been demonstrated that different structures of carbonaceous material can be used to improve the fracture toughness of HAp, as they can easily serve the purpose of being a second phase reinforcement, with the resulting composite still being a biocompatible material. Nanostructured carbonaceous structures, especially those in the form of fibres and sheets, were found to be very effective in increasing the fracture toughness values of HAp. Minor addition of CNTs (3 wt.%) has resulted in a more than 200% increase in fracture toughness of hydroxyapatite-nanorods/CNTs made using spark plasma sintering. This paper presents a current review of the research field of using different carbonaceous materials composited with hydroxyapatite with the intent being to produce high performance biomedically targeted materials.

Development and characterization of a xenograft material from New Zealand sourced bovine cancellous bone.

A xenograft (bovine hydroxyapatite [BHA]) was developed from New Zealand sourced bovine cancellous bone by a successful defatting and deproteinizing procedure. The BHA was chemically, compositionally and structurally characterized. Fourier transform infrared spectroscopy confirmed the removal of organic matter from the bone matrix and the presence of carbonate ( CO32-), hydroxyl (OH- ), and phosphate ( PO43-) functional groups. X-ray diffraction analysis suggested that the processed bone corresponds characteristically to hydroxyapatite (HA). SEM analysis showed that the BHA has an interconnected porous architecture with a pore diameter ranging from 100 to 700 µm while µCT analysis calculated the total porosity as 73.46% ± 1.08. Furthermore, the BHA was stable up to 1000°C and lost only 1.8% of its weight. The Ca/P molar ratio of the BHA was 1.58, which is comparable with commercially available natural HA-Endobon® . After 28 days of incubation in simulated body fluid (SBF), the pH value only fluctuated between 7.1 and 7.5 and the BHA scaffold did not degrade significantly by weight indicating the scaffold had excellent chemical and structural stability. In vitro studies showed the BHA was cytocompatible and supported the proliferative growth of Saos-2 osteoblast cells. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1054-1062, 2017.

Substituted hydroxyapatites for bone regeneration: A review of current trends.

At present hydroxyapatite (HA) is been extensively investigated for biomedical applications, largely as a result of its similarity in composition to the mineral portion of bone. Although HA undergoes osseointegration and is bioactive and osteoconductive, the inherent brittleness and low fracture toughness limits its use under load bearing conditions, also once implanted in the body, HA takes a long time to resorb. The crystal structure of HA is conducive to a variety of ionic substitutions. To accurately mimic the calcium deficient and carbonate-containing nature of HA in bone, both cationic and anionic substituents have been incorporated to synthetic HA. This article focuses on the incorporation of both the well established (Zn, Si, Sr, F, and carbonate) and latest ions (Ag, citrate, iron, niobate, and tantalates) into the HA structure and aims to highlight the key effects of these substitutions in terms of their chemical, physical, and biological properties. It can be shown that a minor substituent cannot only alter the microstructure, stability and crystallinity of the HA structure in an implant, but also have a significant effect on bone cells colonizing the implant, which in turn can influence the new bone formation and bone remodeling processes. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1285-1299, 2017.

Vacuum-assisted infiltration of chitosan or polycaprolactone as a structural reinforcement for sintered cancellous bovine bone graft.

Sintered cancellous bovine bone (SCBB) offers numerous advantages as a bone graft substitute material; however, its mechanical properties require improvement. In this study, SCBB was infiltrated with ε-polycaprolactone (PCL) or chitosan/monetite to improve mechanical properties while retaining valuable SCBB structure. Organic infiltrating solutions consisted of (i) chitosan and monetite (CaHPO(4)) dissolved in hydrochloric acid; (ii) chitosan, monetite (CaHPO(4)), and genipin dissolved in hydrochloric acid; or (iii) PCL polymer dissolved in tetrahydrofuran (THF). Porous SCBB materials were infiltrated with one of the three solutions using vacuum-assisted infiltration. Chitosan and CaHPO(4) were immobilized in the SCBB structure by reprecipitation following a pH increase in an ammonia atmosphere. Genipin was used in one sample group to immobilize chitosan via crosslinking. PCL was immobilized by evaporating the THF carrier solvent. Mechanical compression testing showed an improvement in ultimate stress for the SCBB with chitosan/CaHPO(4) infiltrates, whereas PCL samples showed an increase in modulus. All SCBB samples were found to demonstrate favorable in vitro biocompatibility when subjected to L929 mouse fibroblast cells but required a vigorous washing regime to eradicate toxic ammonia residue. In conclusion, infiltration of SCBB with a polymeric or organic/mineral composite matrix positively modifies SCBB mechanical properties in favor of bone grafting applications.

Synthesis and characterization of polymer-protected rhodium and palladium sols in mixed media.

A series of polymer-protected rhodium and palladium sols were generated in mixed (organic/aqueous) dispersion media. Protecting polymers used were PAA, PVP, PVA and an oligosaccharide-based protecting agent, arabinogalactan. This is the first systematic study of colloids generated in mixed solvent dispersion media with these protecting agents. The sols were characterized by TEM, UV/Vis spectrophotometry and Photon Correlation Spectroscopy (PCS). In general, the macroscopic stability of the sols depended on the protecting agent/organic cosolvent combination used, with a nonlinear dependence on cosolvent concentration. Observed sol stabilities correlated well with pre-existing hydrodynamic data for the protective polymers, e.g. radius of gyration data. Average particle size and polydispersity of all sols examined decreased as cosolvent concentration increased, with actual cosolvent concentration, protecting agent and reducing agent used influencing the extent of decrease. Sol particle sizes ranged from 0.8 nm to 6.0 nm with sol turbidities decreasing in parallel with the particle sizes. Hydrodynamic sizes (from PCS) tended to reflect the sols' macroscopic stabilities. FTIR studies using adsorbed CO as a spectroscopic surface probe indicated decreases in the ν(CO)(ads) stretching frequency with an increase in cosolvent concentration due to competitive adsorption of CO with the cosolvent molecules on the metal colloid surfaces.

The application of co-melt-extruded poly(ε-caprolactone) as a controlled release drug delivery device when combined with novel bioactive drug candidates: Membrane permeation and Hanson dissolution studies.

Eight bioactive drug compounds (abamectin, amoxicillin, dexamethasone, dexamethasone valerate, ketoprofen, melatonin, oestradiol 17ß, and oestradiol benzoate) were combined via melt extrusion and disc pressing processes with a polycaprolactone (PCL) matrix and were then evaluated and compared via membrane diffusion and Hanson dissolution studies. This investigation was to determine the potential of this matrix to act as a controlled release drug delivery vehicle for a number of drugs not previously combined with PCL in a melt extrusion mix. The inclusion of the progesterone/PCL system, for which the drug release behaviour has been well studied before was intended for comparison with the PCL systems incorporating drugs that have received little research attention in the past. Initial studies centred on an evaluation of the permeation ability of the bioactive drugs dissolved in aqueous cyclodextrin solutions through a poly(ε-caprolactone) (PCL) membrane using Valia-Chien side-by-side cells. Permeation rates were mostly low and found to range from 0 to 122 µg h(-1) with only ketoprofen, melatonin, and progesterone displaying rates exceeding 20 µg h(-1). Hanson dissolution release profiles in aqueous alcohol were subsequently measured for the 9 melt extruded PCL/drug combinations and led to Hanson release rates of 0-556 µg cm(-2) h(-0.5) with dexamethasone, dexamethasone valerate, ketoprofen, melatonin, and progesterone giving values exceeding 100 µg cm(-2) h(-0.5). A number of drugs such as the dexamethasones probably performed better than they did in the permeability rate measurements because of the less polar aqueous alcoholic solvent used. In searching for useful correlations between the drug physicochemical properties and release rate, only a moderate correlation (R (2)=0.5675) between Hanson dissolution release rate and permeation rate was found. This suggests that the release rate and the permeation are both controlled by the rate of drug diffusion through the PCL with release rate involving an additional dissolution process (of the drug) before permeation occurs accounting for the moderate correlation. In general, of the eight drugs considered, it was clear that the oestradiol-based drugs, abamectin, and amoxicillin were generally not suited to drug delivery via PCL under the conditions used. However, ketoprofen was found to be very suitable as a drug candidate for melt extrusion with PCL with dexamethasone valerate, dexamethasone, and melatonin also showing potential as candidates though to a much lesser extent.

Phosphorylated, cellulose-based substrates as potential adsorbents for bone morphogenetic proteins in biomedical applications: a protein adsorption screening study using cytochrome C as a bone morphogenetic protein mimic.

Screening studies aimed at identifying useful biomedical materials that (when combined with implants) can attract bone morphogenetic proteins to their surfaces have been conducted. In this paper, the screening process has involved carrying out protein adsorption studies using cytochrome C, as a BMP protein mimic on phosphorylated cellulose-based substrates. These studies have shown that phosphorylation of cellulose produces materials that are capable of attracting the adsorption of cytochrome C to their surface. In contrast, negligible cytochrome C adsorption was observed on the unphosphorylated cellulose-based materials. The selective uptake of the positively charged cytochrome C (from solutions at pH 9.51) by the negatively charged phosphorylated cotton and microcrystalline cellulose substrates was primarily due to this protein's high isoelectric point (i.e.p) of 9.8 which gives it a positive charge at pH

In situ characterisation of the aqueous gold colloid interface using CO as a surface probe: IR spectroscopic studies.

Fourier transform infrared (FTIR) spectra are presented of CO gas-treated protected gold colloids prepared from hydrazinium hydrate reduction of an Au(III) precursor which reproducibly feature a weak, shortlived peak at ca. 2169 cm(-1). When the gold colloid was treated with 99% isotopically enriched (13)CO gas, the IR peak shifted to a frequency of 2114 cm(-1) which indicated that it represented a simple gold monocarbonyl species. The value of 2169 cm(-1) for the CO stretching frequency suggests the peak represents CO physisorbed on oxidised gold atoms on the colloid surface. The peak is not observed when the concentration of the colloidally dispersed gold is reduced either by use of lower starting salt concentrations or by aggregation. It is also not observed when solutions of the protecting agent or reducing agent or the dispersion medium (water) or even the starting Au(III) salts are CO-treated individually. This confirms that the spectral feature is uniquely associated with colloidally dispersed gold. In general, the work has shown that the surfaces of Au colloids in situ have partially oxidised Au character which is of interest in systems where supported nanoparticulate gold derived from colloid preparations are considered for low temperature oxidation catalysts for CO.

Growth of calcium hydroxyapatite (Ca-HAp) on cholesterol and cholestanol crystals from a simulated body fluid: A possible insight into the pathological calcifications associated with atherosclerosis.

An experimental study into calcium phosphate (CP) nucleation and growth on cholesterol and cholestanol surfaces from a supersaturated simulated body fluid (SBF) is presented with the overall aim of gaining some fundamental insights into the pathological calcifications associated with atherosclerosis. Soaking of pressed cholesterol disks at physiological temperature in SBF solutions was found to lead to CP nucleation and growth if the disks were surface roughened and if an SBF with concentrations of the calcium and hydrogen phosphate ions at 2.25x physiological concentrations was used. The CP phase deposited was shown via SEM micrographs to possess a florette type morphology akin to that observed in earlier reported studies. The use of recrystallised cholesterol and cholestanol microcrystals as substrates for soaking in SBF facilitated the observation of CP deposition. In general, cholesterol recrystallised from polar solvents like 95% ethanol as a cholesterol monohydrate phase which was a better substrate for CP growth than cholesterol recrystallised from more non-polar solvents (e.g., benzene) which produced anhydrous cholesterol phases. CP was also observed to form on recrystallised cholestanol microcrystals, a molecule closely related to cholesterol. Inductively coupled plasma optical emission spectrometry (ICP-OES) data gave confirmation that Ca:P mole ratios of the grown CP were 1.3-1.5 suggesting a mixed phase of octacalcium phosphate (OCP) and Ca-deficient HAp and that the CP coating grows (with time of soaking) on the substrates after nucleation in the SBF growth medium. Infrared (IR) spectra of the extracted coatings from the cholesterol substrates confirmed that the CP phase deposited is a semi crystalline HAp with either carbonate substituted into its structure or else co-deposited as calcium carbonate. Soaking experiments involving modified cholesterol substrates in which the OH group in the molecule was replaced with the oleiyl or phosphonate group showed no CP nucleation and growth. This observation illustrates the importance of the known epitaxial relationship between cholesterol and HAp (which theoretically predicts favourable deposition of one phase upon the other) and the consequences of its destruction (by chemical modification of the cholesterol). In the case of the phosphorylated cholesterol, failure of this substrate to nucleate CP phases may have also been caused by the reduction in concentration of free solution Ca2+ in the SBF medium by complexation with the phosphonate groups on the phosphorylated cholesterol. This would have reduced the ion product of Ca2+ and inorganic phosphate and lowered the degree of supersaturation in the SBF medium.

The evaluation of processed cancellous bovine bone as a bone graft substitute.

OBJECTIVE: To determine the ability of a novel bovine cancellous bone xenoimplant to act as an osteoconductive graft in an ovine femoral defect model. An autograft harvested from the xenoimplant site was placed in a contralateral limb defect for comparison. MATERIAL AND METHODS: The xenoimplant used had been rendered immunologically inert by a novel defatting and deproteinating process. Following surgical implantation of cores into condylar cancellous bone defects, fluorochrome labels were administered to 12 sheep at 2 1/2, 4 1/2 and 8 weeks. Incorporation of the xenoimplants and autografts into the host bone was compared radiographically and histomorphometrically at 10 weeks. RESULTS: Radiographically, the degree of osteointegration was comparable. Histomorphometric data, consisting of labelled surface (LS) estimates, confirmed osteoconductive properties of both the xenoimplants and autografts. Remodelling activity was greatest in the xenoimplants at 2 1/2, weeks. At 4 1/2 weeks, there was more activity in the autograft, but by 8 weeks they were performing similarly. Xenoimplant-LS estimates were comparable or greater than those of the autograft at all times. CONCLUSIONS: Processed bovine cancellous bone xenoimplants were osteoconductive in this model and show promise for development as a biomaterial in human and veterinary orthopaedic surgery.

In situ infrared spectroelectrochemical studies of the corrosion of a nickel electrode as a function of applied potential in cyanate, thiocyanate, and selenocyanate solutions.

This paper presents the first subtractively normalized interfacial Fourier transform infrared spectroscopic (SNIFTIRS) study of the corrosion system Ni/XCN(-) (X=O, S, Se), pH 11, [XCN(-)]=0.05 molL(-1), supporting electrolyte 0.1 molL(-1) KNO(3), for a nickel electrode as a function of applied potential. Cyclic voltammograms, in situ infrared spectra, and current-potential data (recorded while the infrared spectral acquisition was in progress) were recorded for a nickel electrode in a three-electrode thin-layer cell containing the pseudohalides OCN(-), SCN(-), or SeCN(-) ions at pH 11 in a supporting electrolyte of KNO(3). In general, the data showed that all of the pseudohalide ions studied caused corrosion of the nickel electrode by forming the respective nickel-pseudohalide complex ion species as the potential was stepped anodically. Two of the ions, SCN(-) and SeCN(-), caused surface modifications to the electrode which influenced the electrochemical reactions with respect to CO(2) formation. The Ni/SeCN(-) system, for instance, exhibited signs of instability during the spectroelectrochemical experiment, red-brown coatings observed on the electrode caused by the decomposition of the selenocyanate ion to colloidally dispersed elemental selenium. The selenium coated the electrode, hence modifying the surface and consequently the electrochemistry, by causing the "early" appearance of CO(2)-associated IR peaks in SNIFTIRS spectra recorded from the electrode system at potentials lower than those for the Ni/OCN(-) system. In contrast, CO(2) formation at the electrode surface was not observed in the Ni/SCN(-) system, which was likely to have been caused by nickel sulfide poisoning of the electrode surface. In the Ni/SCN(-) and Ni/SeCN(-) systems, IR spectra also indicated the buildup of Ni(SCN)(2) and Ni(SeCN)(2) salts in the thin layer by the appearance of a peak at ca. 2165 cm(-1) at anodic values of the applied potential.

Rhenium-Based Hydrosols: Preparation and Properties.

Transmission electron microscope (TEM), ultraviolet-visible (UV/vis), electrospray mass spectrometric (ESMS), and X-ray photoelectron spectroscopic (XPS) studies are presented on a little studied rhenium hydrosol system produced by reduction of aqueous K(2)ReCl(6) with hydrazine in the presence or absence of a gum arabic protecting agent. The studies indicate that hydrazine-generated "rhenium hydrosols" are unstable in water and slowly dissolve over time in aqueous media to form the highly stable perrhenate ion. Copyright 2001 Academic Press.