Rapid fabrication of bio-inspired, mineralized polysaccharide coatings

Bio-inspired, mineralized polysaccharide coatings consisting of interspaced alginate–chitosan and calcium phosphate carbonate planar composite films were rapidly fabricated via electrostatic layer-by-layer (LBL) self-assembly and organic-matrix-mediated nucleation and growth. Experimental design, a fractional factorial design (2^8-4), was used to identify the optimum LBL film process parameters for fast film growth. Calcium phosphate carbonates rapidly precipitated on the fast growing LBL films from a supersaturated solution in around 15 min. The number of bilayers of LBL films and soaking time in calcium phosphate carbonate solution were found to affect the morphology of calcium phosphate carbonates, indicating a complex, organic matrix mediated mineralization process. The coating protocol in the current study was readily transferable to a variety of template geometries and materials. The resulting planar composite coatings could be applied for the modification of orthopaedic and periodontal implant surfaces and also for controlled growth of hematopoietic or mesenchymal stem cells.     

Calcium phosphate thin film processing by pulsed laser deposition and in situ assisted ultraviolet pulsed laser deposition

Calcium orthophosphates (CaP) and hydroxyapatite (HA) were intensively studied in order to design and develop a new generation of bioactive and osteoconductive bone prostheses. The main drawback now in the CaP and HA thin films processing persists in their poor mechanical characteristics, namely hardness, tensile and cohesive strength, and adherence to the metallic substrate. We report here a critical comparison between the microstructure and mechanical properties of HA and CaP thin films grown by two methods. The films were grown by KrF* pulsed laser deposition (PLD) or KrF* pulsed laser deposition assisted by in situ ultraviolet radiation emitted by a low pressure Hg lamp (UV-assisted PLD). The PLD films were deposited at room temperature, in vacuum on Ti–5Al–2.5Fe alloy substrate previously coated with a TiN buffer layer. After deposition the films were annealed in ambient air at 500–600 °C. The UV-assisted PLD films were grown in (10^–2–10^–1 Pa) oxygen directly on Ti–5Al–2.5Fe substrates heated at 500–600 °C. The films grown by classical PLD are crystalline and stoichiometric. The films grown by UV-assisted PLD were crystalline and exhibit the best mechanical characteristics with values of hardness and Young modulus of 6–7 and 150–170 GPa, respectively, which are unusually high for the calcium phosphate ceramics. To the difference of PLD films, in the case of UV-assisted PLD, the GIXRD spectra show the decomposition of HA in Ca₂P₂O₇, Ca₂P₂O₉ and CaO. The UV lamp radiation enhanced the gas reactivity and atoms mobility during processing, increasing the tensile strength of the film, while the HA structure was destroyed.     

Diffusion galvanizing of brass

Diffusion galvanizing of brasses (L-62, L-68, and LS-59-1) offers a means of producing zinc-rich diffused coatings whose microhardness is 3–4 times higher than that of a given brass. The wear resistance of surface coatings of this kind (measured against a hard steel disk under pressures of 30–450 g/mm² and at a circular speed of 1.2 m/sec) is 5–10 times higher than that of uncoated brasses.     

Properties of calcium phosphate coatings deposited and modified with lasers

Physical, chemical and biological properties of calcium phosphate coatings fabricated by a pulse laser deposition method at room temperature (RT PLD) have been studied. In vitro evaluation of RT PLD coatings on bioresorbable polymers (Poly--caprolactone and Poly-L-lactide) have been carried out. It was shown that both polymers support osteoblast growth, with increased cell activity, alkaline phosphatase activity and total protein content on those surfaces that have been coated. The advantages of RT PLD coatings in biomaterials surface optimization are discussed.     

Formation of zinc phosphate coatings on AA6061 aluminum alloy

Conditions for forming zinc phosphate conversion coatings on AA6061 aluminum alloy have been investigated by characterizing coatings formed for different parameters of the coating bath. Morphological and compositional information on the coatings was assessed by SEM, EDX and XPS, and simple adhesion tests were undertaken to indicate the strengths of coating attachment. The emphasis was to identify conditions that give high coverage, uniform coatings of small, strongly adhered, zinc phosphate crystals. The use of low-zinc solutions (e.g. an atomic Zn/P ratio of 0.07) and normal-zinc solutions (Zn/P ratio 0.25) were compared; coatings formed by the two solution types appear comparable at pH 2, although at pH 4 the low-zinc solution is more effective. Fluoride in the concentration range 200–400 ppm is indicated to be a useful additive for the normal-zinc coating bath and in the 600–1000 ppm range for the low-zinc process. The use of acid etching in the pre-treatment appears to yield better coatings than when mechanical polishing alone is used.     

Preparation and characterization of magnesium/carbonate co-substituted hydroxyapatites

A new synthesis/processing method has been devised to produce magnesium/carbonate co-substituted hydroxyapatite ceramics that do not decompose to tricalcium phosphate (TCP) on sintering. Using this method, a series of magnesium/carbonate co-substituted hydroxyapatite (Mg/CO₃–HA) compositions, containing between 0 and 0.35 wt % Mg and approximately 0.9 wt % CO₃ were prepared. Sintering the Mg/CO₃–HA compositions in a CO₂/H₂O atmosphere yields a single crystalline phase that appears to be identical to stoichiometric HA. In contrast, when the compositions were prepared in the absence of carbonate and were sintered in air, the phase composition was a biphasic mixture of HA and TCP e.g. for 0.25 wt % Mg substitution the phase composition was approximately 60%HA/40% TCP. Clearly, both the synthesis route and the processing (i.e. sintering) route are of importance in the production of a single-phase Mg/CO₃–HA ceramic. Fourier transform infrared (FTIR) spectroscopy has indicated that the Mg/CO₃–HA ceramics still contained carbonate groups after sintering at 1200 °C. Chemical analysis by X-ray fluorescence spectroscopy (XRF) and C–H–N analysis has shown that the cation/anion molar ratio (i.e. [Ca+Mg]/[P+C/2]) of the different compositions were 1.68(±0.01), which is equivalent to the Ca/P molar ratio of stoichiometric HA. Although the magnesium/carbonate co-substitution had a positive effect in preventing phase decomposition during sintering, it appeared to have a negative effect on the densification of the MgCO₃–HA ceramics, compared to stoichiometric HA.     

Formation of carbonate apatite on calcium phosphate coatings containing silver ions

The prerequisite for bioactive materials to bond to living bone is the formation of biologically equivalent carbonate apatite on their surfaces in the body. Calcium phosphate ceramic surfaces can be transformed to a biological apatite through a series of surface reactions including dissolution–precipitation and ion exchange. In the present work, apatite coatings with different crystallinity, compositions and crystal sizes, including a well-crystallized hydroxyapatite coating, were synthesized electrochemically and doped with silver ions in silver nitrate solution at room temperature. The formation of a new carbonate apatite on the surface of these coatings was investigated in an acellular simulated body fluid with ion concentrations comparable with those of human blood plasma, using scanning electron microscopy and Fourier transform-infrared spectroscopy. The results show that small quantities of silver ions incorporated into apatite coatings may have a strong stimulatory effect on the formation of carbonate apatite without adversely affecting the chemical stability of these coatings.     

A simple method to prepare calcium phosphate coatings on Ti6Al4V

A two-step chemical treatment followed by immersion in a supersaturated calcification solution (SCS) was found to be a simple way to prepare calcium phosphate (Ca–P) coatings on Ti6Al4V. The Ca–P deposition on the treated metallic surfaces could be accelerated by employing a pre-calcification (Pre-Ca) procedure prior to immersion in SCS. The two-step treatment was performed by etching the metallic plates with a mixture of HCl and H2SO4 followed by ageing in boiling diluted NaOH solution at 140°C. Pre-Ca was carried out by incubating the two-step treated plates in Na2HPO4 solution and then in saturated Ca(OH)2 solution. The formation of a bioactive microporous surface oxide layer on Ti6Al4V by the two-step treatment was most probably responsible for the induction of Ca–P precipitation. The deposition rates and compositions of Ca–P coatings in two different SCSs were investigated by means of scanning electron microscopy, X-ray diffraction and infrared spectrophotometry.     

Control of phase composition in hydroxyapatite/tetracalcium phosphate biphasic thin coatings for biomedical applications

Biphasic calcium phosphates comprising well-controlled mixtures of nonresorbable hydroxyapatite and other resorbable calcium phosphate phases often exhibit a combination of enhanced bioactivity and mechanical stability that is difficult to achieve in single-phase materials. This makes these biphasic bioceramics promising substrate materials for applications in bone tissue regeneration and repair. In this paper we report the synthesis of highly crystalline, biphasic coatings of hydroxyapatite/tetracalcium phosphate with control over the weight fraction of the constituent phases. The coatings were produced by pulsed laser deposition using ablation targets of pure crystalline hydroxyapatite. The fraction of tetracalcium phosphate phase in the coatings was controlled by varying the substrate temperature and the partial pressure of water vapor in the deposition chamber. A systematic study of phase composition in the hydroxyapatite/tetracalcium phosphate biphasic coatings was performed with X-ray diffraction. Tetracalcium phosphate in the coatings obtained at high substrate temperature is not formed by partial conversion of previously deposited hydroxyapatite. Instead, it is produced by nucleation and growth of tetracalcium phosphate itself from the ablation products of the hydroxyapatite target or by accretion of tetracalcium phosphate grains formed during ablation. This finding was confirmed by formation of calcium oxide, not tetracalcium phosphate, after annealing of pure hydroxyapatite coatings at high temperatures of 700–850∘C.     

Further studies of calcium phosphate growth on phosphorylated cotton fibres

Further studies using scanning electron microscopy/energy dispersive X-ray analysis (SEM/EDX), micro-Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS) and solid state magic angle spinning nuclear magnetic resonance (MAS NMR) techniques of calcium phosphate growth on Ca(OH)₂-treated urea/H₃PO₃- and urea/H₃PO₄-modified cotton fibres are reported. In the case of the Ca(OH)₂-treated urea/H₃PO₃-modified fibres which have been reported in an earlier paper, further experiments subjecting the urea/H₃PO₃-modified cotton to alternative soaking treatment procedures to Ca(OH)₂ as well as different calcium phosphate growth media such as the alkaline phosphatase-catalysed hydrolysis of disodium p-nitrophenylphosphate to free phosphate have reaffirmed the importance of the Ca(OH)₂ treatment step for the stimulus and growth of calcium phosphate growth on the fibres. Studies on cotton phosphorylated by a slightly different method using urea/H₃PO₄ instead of urea/H₃PO₃ show that a phosphorylated cotton with similar properties to the urea/H₃PO₃-modified fibres can be produced. Soaking of these fibres in saturated Ca(OH)₂ solution leads to cotton coated with thin layers of calcium phosphate formed by partial hydrolysis of the PO₄ functionalities in the phosphorylated cotton which are believed to act as nucleation layers for further calcium phosphate deposition when the fibres are subsequently soaked in 1.5×SBF solution. SEM/EDX studies of the calcium phosphate coatings formed on the Ca(OH)₂-treated urea-H₃PO₄ fibres as a function of soaking time in 1.5 × SBF show that coatings deposit and become noticeably thick after approximately 9 days. XPS studies indicated the presence of carbonate species in the calcium phosphate coating deposited. In common with the calcium phosphate coated Ca(OH)₂-treated urea/H₃PO₃-modified fibres studied earlier, the average EDX-measured Ca: P ratios of the coatings formed on the Ca(OH)₂-treated urea/H₃PO₄ fibres are 1.60 and give very similar micro-FTIR spectra with evidence of carbonate which suggests that amorphous calcium deficient apatite has deposited.     

Sol-gel-derived hydroxyapatite powders and coatings

Hydroxyapatite (HAP) and tri-calcium phosphate (TCP) powders and coatings with a Ca/P molar ratio from 1.56 to 1.77 were prepared by the sol-gel technique using calcium 2-ethylhexanoate (Ca(O₂C₈H₁₅)₂) and 2-ethyl-hexyl-phosphate as calcium and phosphorus precursors, respectively. The structural evolution and phase formation mechanisms of HAP and tri-calcium phosphate in calcined powders and coatings on Si wafer and Ti-alloy substrates (Ti-30Nb-3Al and Ti-5Al-2.5Fe) were characterized by X-ray diffraction, Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). The elimination of organics was studied by differential thermal analysis (DTA) and thermogravimetry (TGA). Two different formation mechanisms of crystallization are proposed. In sols with Ca/P 1.67, -tricalcium phosphate is formed as the major phase and hydroxyapatite as a minor phase by calcination at 700°C. At 900°C these phases react to form AB-type carbonated hydroxyapatite (Ca_10–2x/3[(PO₄)_6–x (CO₃) _x ][(OH)_2–x/3–2y (CO₃) _y ]). A release of CO₂ substituting PO₄ ^3– occurs between 900°C and 1100°C yielding carbonate apatite, Ca₁₀(PO₄)₆[(OH)_2–2y (CO₃) _y ], whereas CO₂ substituting OH^– groups in the apatite structure is released above 1200°C. In sols with Ca/P 1.70, rather than carbonate apatite, B-carbonated hydroxyapatite Ca_10–2x/3[(PO₄)_6–x (CO₃) _x ](OH)₂ is formed, which subsequently decomposes into HAP and CaO above 1200°C. The optimum sintering conditions for coatings on Ti-alloys are found to be 600°C for 10 minutes, since, at higher temperature, oxidation of titanium and the formation of rutile (TiO₂) occur. Dip coating and sintering in two cycles yielded a homogeneous and dense coated film with a thickness of 250 nm.     

Adhesion study of pulsed laser deposited hydroxyapatite coating on laser surface nitrided titanium

Hydroxyapatite (HA) coatings were fabricated by pulsed laser deposition (PLD) on commercially pure titanium which had been subjected to different types of pre-treatment. These include: (i) 60-grit SiC grinding, (ii) 320-grit SiC grinding, (iii) 1-µm diamond paste mirror-finishing, (iv) etching with Knoll solution, and (v) laser surface nitriding followed by selective etching. The HA coatings were pulsed laser deposited at different water-vapor pressures to determine the optimal processing conditions. The nitrided-etched specimen exhibits a three dimensional TiN dendritic network which promotes the adhesion between HA coating and titanium substrate. Among the specimens with different pre-treatments, the adhesion strength of HA is the highest for the nitrided-etched specimen, reaching about twice that for the mirror-finished specimen. Thin-film X-ray diffraction shows a high degree of crystallinity for the PLD deposited HA. According to energy-dispersive X-ray analysis, the Ca/P ratio of the deposited HA reaches an approximate value of 1.7, similar to that of the HA target. Scanning-electron microscopy reveals that the deposited HA is about 4 μm in thickness. Growth of apatite was rapidly induced on the HA coated specimens when immersed in Hanks' solution for 4 days, indicating that the PLD HA coating is highly bone bioactive. This could be partly due to the high wettability of the PLD HA surface.     

Growth and structure of TiCxNy coatings chemically vapour deposited on graphite substrates

TiCxNy coatings were grown on graphite substrates in a computer-controlled, hot-wall chemical vapour deposition (CVD) reactor, using gas mixtures of TiCl4–CH4–N2–H2 at a total pressure of 10.7 kPa (80 torr) and at a temperature of 1400 K. Growth rate, composition, morphology and crystallographic texture of the TiCxNy coatings were investigated as a function of the CH4/CH4+N2 ratio in the range 0–1 at a constant CH4+N2 flow rate of 370 standard cubic centimeters per minute (sccm). The C/C+N ratio and growth rate of the TiCxNy coatings increased with increasing CH4/CH4+N2 ratio in the gas phase. The compositions of the coatings with C/C+N ratios in the range 0–1 were found to be between the thermodynamic and the kinetic predictions. Morphology and preferred orientation of the coatings were observed to be strongly affected by the CH4/CH4+N2 ratio in the gas phase.     

Microstructural characterization of hydroxyapatite coating on titanium

The microstructure of hydroxyapatite plasma sprayed onto titanium alloy has been studied by using transmission electron microscopy. It has been shown that while substantial portions of the coating are crystalline hydroxyapatite, regions of amorphous calcium phosphate with Ca/P ratios of 0.6–1.0 are also present, both in the coatings and at the metal-ceramic interface. The microstructures observed have also been found to be consistent with devitrification of the amorphous calcium phosphates producing regions of very fine grained hydroxyapatite. A calcium titanate phase has also been detected at the metal-ceramic interface produced by the chemical reaction of hydroxyapatite to titanium.     

High-Pressure Measurements of the Viscosity and Density of Two Polyethers and Two Dialkyl Carbonates

The viscosity and density of four pure liquid compounds (dimethyl carbonate, diethyl carbonate, triethylene glycol dimethyl ether, and tetraethylene glycol dimethyl ether) were measured at several temperatures between 283.15 and 353.15 K. The density measurements were performed up to 60 MPa with an uncertainty of 1×10^–4g·cm^–3. The viscosity at atmospheric pressure was measured with an Ubbelohde-type glass capillary tube viscometer with an uncertainty of ±1%. At pressures up to 100 MPa the viscosity was determined with a falling ball viscometer with an uncertainty of ±2%. The density (410 experimental values) and viscosity data (184 experimental values) were fitted to several correlation equations.     

Structural investigations of phosphate glasses: a detailed infrared study of the x(PbO)-(1−x) P2O5 vitreous system

The results and detailed discussion of an extensive experimental study of infrared spectra of the x (PbO)-(1–x)P₂O₅ vitreous system (x=0.3–0.75) together with a brief review of infrared spectra of phosphate compounds, are presented. Theoretical models employed in the interpretation of infrared spectra of glasses have been reviewed. The frequency ranges of various infrared bands belonging to PO ₄ ^3– and P₂O ₇ ^4– , observed in different phosphate compounds, are discussed. The glassy and quenched samples were prepared from PbO and NH₄H₂PO₄ by the rapid quenching technique. The infrared spectra of the constituents of the system, PbO and P₂O₅, in their polycrystalline and glassy forms, have been discussed. The intensity and wavenumbers of the infrared bands around 1600 and 3300 cm^–1, assigned to the bending and stretching modes in H₂O trapped by the hygroscopic glasses, have been followed for different compositions with x<0.5. The changes observed in these infrared bands established the role of water as an additional glass modifier. The intensity and frequency variations of the infrared bands have been followed through all the compositions for characteristic phosphate group frequencies including P=O, P-O-P stretching and bending modes and P-O bending mode. The results clearly suggest that the x(PbO)-(1–x)P₂O₅ system undergoes gradual structural changes from metaphosphate (x=0.5), to pyrophosphate (x=0.66) and to orthophosphate (x=0.75). The continuing presence of the infrared band, in varying intensity, in the region 1200–1280 cm^–1 attributed to P=O, suggests that the glass-forming ability of the binary system is extendable at least up to x=0.66 composition, and that no complete rupture of P=O bond by Pb²⁺ takes place. The ionic character of the phosphate groups, P-O^(–), PO ₄ ^3– is well revealed by significant changes with the PbO content in the spectral features of the infrared bands around 1120 and 980 cm^–1 respectively. The maximum intensity of the P-O^(–) band at 1120 cm^–1 for 55 mol% PbO suggests a partial breakdown of the covalent vitreous network of the phosphates and formation of a crystalline phase consisting of ionic groups PO ₄ ^3– , P₂O ₆ ^2– and P₂O ₇ ^4– for PbO greater than 55 mol%. The observed pattern of variation in the intensity of the infrared bands in the 940–1080 cm^–1 region attributed to the v₃-mode in PO ₄ ^3– , suggests a gradual transformation of PO ₄ ^3– units to PO ₃ ^– groups in lead meta-phosphate glass and then their restoration to PO ₄ ^3– groups of pyro- and ortho-phosphate quenched samples. The results indicate a gradual decrease in the number of bridging oxygens and increase in the resonance behaviour of non-bridging oxygens as the mole percentage of metal oxide (PbO) increases in the glass. The infrared spectra of several binary phosphate glasses have been reviewed in the context of the study of effect of the cation on the infrared spectra. It is found that the influence of the cation on the infrared spectra of phosphate glasses does not show any striking regularity. Theoretical calculations of these band frequencies were found to agree well only in the case of pure stretching (P=O and O-H) vibrations and pure bending (P-O-P and O-H) vibrations. The disagreement in the case of P-O^(–), P-O-H and other modes of P-O-P groups, has been attributed to the mixed nature of modes occurring in glasses. The changes in the positions of the characteristic bands and their relative intensities are strongly dependent on the structural units and PbO content in the phosphate glasses and the results emphasize the role of PbO as a network modifier.     

Synthesis of TiO2 and TiN nanosize powders by intense light ion-beam evaporation

Ultrafine nanosized powders of TiO2 and TiN have been synthesized by the reactive ion-beam evaporation technique. In an oxygen or nitrogen atmosphere (1–10 Torr), a high-power-density ion beam (about 0.4 GW cm-2) is focused onto a Ti target; as a result of the interaction between ablated particles and gas, fine and ultrafine powders are synthesized. The influence of gas pressure and target–collector distance on the particle composition, size distribution and shape have been studied. At longer target–collector distances and higher pressures, powders in the 4–45 nm range are collected while, at shorter distances and lower pressures, the powders are in the micrometre range. TiO2 particles are spherical in shape, while TiN particles are cubic.     

Biological effects of aluminium diffusion from plasma-sprayed alumina coatings

Plasma-sprayed alumina (Al₂O₃) coatings on metal stems of hip prostheses are used to favour bone apposition on the stem without fibrous interposition. We tested both in vitro and in vivo in rabbits, alumina coatings in order to evaluate the biological effect of this material on bone. Mice fibroblasts were grown on Al₂O₃-coated discs and time course of aluminium concentration was recorded in two phosphate and citrate buffers (pH 4 and 7) bathing the alumina coated discs. Alumina-coated cylinders were implanted into femur condyles of ten rabbits for periods of time from 1–6 months. Then, they were histologically analysed using light and scanning electron microscopy, and X-ray microanalysis. Cell proliferation was not affected on alumina coatings compared to controls. In pH 4 buffer, aluminium was released from the coatings. From a period of implantation of 4–6 months an increasing demineralization process took place in the bone at the coating contact. Aurine staining showed the presence of aluminium at the interface between the non-mineralized and the mineralized bone. These results suggest that aluminium is released from alumina coatings and leads to bone demineralization at the coating contact.     

Investigation of High-Velocity Suspension Flame Sprayed (HVSFS) glass coatings

High-Velocity Suspension Flame Spraying (HVSFS) has recently emerged as a potential alternative to conventional HVOF-spraying: employing liquid suspensions instead of dry powder feedstock enables the use of very fine grain-sized particles, resulting in small-sized lamellae. Thin, low-porosity coatings can thus be manufactured. This paper details the first attempt at manufacturing glass coatings using the HVSFS technique: these coatings can have multiple applications (anti-corrosion coatings on metal and ceramic substrates, bio-compatible coatings, etc). A CaO–ZrO₂–SiO₂ glass frit was selected for this attempt. Excellent potentialities emerged (very low porosity), but some problems still existed (big “droplet-like” features on the coating surface), which have recently been largely overcome thanks to process modifications.     

Structural–Thermodynamic Model for Synergistic Exchange of Hydroxyl and Fluoride Ions in Apatites with the Participation of the Carbonate Ion

Based on crystal-chemical and thermodynamic data, a model was proposed for the synergistic participation of the carbonate ion in hydroxyl–fluoride ion exchange in the Ca channels of apatites (bony and dental tissue). The synergistic effect of the carbonate ion develops upon partial isomorphic substitution [PO₄] groups and consists in equalizing the bonding energies of F^– and OH^– in the Ca channel.