Improvement of bonding strength between biomimetic apatite coating and substrate

Bone-like apatite coatings were prepared using a biomimetic method in a modified simulated body fluid (m-SBF). The effect of the m-SBF volume on the apatite coating quality was studied. Three m-SBF volumes, 50, 100, and 200 mL, were employed to immerse titanium substrates in a sealed container so as to produce apatite coatings with different properties, namely types I, type II, and type III apatite coatings, respectively. The coatings were characterized using X-ray diffraction and environmental scanning electron microscope. The bonding between the coating and the Ti substrate was evaluated using an adhesive strength test. All three apatite coatings demonstrated a poorly crystallized structure, and the coatings formed exhibited a uniformed surface morphology. Further increasing the m-SBF volume, small globules of apatite started to form on the surface of the coating. The bonding strength for the three coating systems were 8.52 +/- 2.41, 10.36 +/- 2.78, and 17.23 +/- 2.55 MPa for types I, II, and III apatite coatings, respectively. The failure analyses suggested that type III coating failed mostly at the interface between the coating and the substrate, while type I and II coatings failed mostly within the apatite coating. Our study revealed that a dense, thick, well-adhered apatite coating could be achieved by carefully controlling the volume of m-SBF.     

Transformation of nacre coatings into apatite coatings in phosphate buffer solution at low temperature

Nacre coatings were deposited on Ti6Al4V substrates by electrophoretic technique, and subsequently converted into apatite coatings with hierarchical porous structures by treatment with a phosphate buffer solution. The samples were characterized by X-ray diffraction, Fourier transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy, inductively coupled plasma optical emission spectroscopy, X-ray photoelectron spectroscopy (XPS), and N(2) adsorption-desorption isotherms. The results show that the nacre coatings are converted into the plate-like apatite coatings via a dissolution-precipitation reaction, while the organic components of the nacre are reserved. The mesopores with pore size of 4.4 nm are formed within the plate-like structure, and the macropores are formed among the plate-like structure. Simulated body fluid (SBF) immersion tests reveal that the apatite coatings have a good in vitro bioactivity. Bone-like apatite crystals are formed on the surfaces of the apatite coatings after soaking in SBF for 12 h, and fill up the macropores on the coatings with increasing the soaking time. In addition, XPS indicates that a TiO(x) layer and PO(4) (3-) ions appear on the substrate surfaces by pretreatment with a H(3)PO(4)/HF solution. The TiO(x) layer and PO(4) (3-) ions can induce the formation of apatite crystals, resulting in a composition gradient from the oxide layer to the external apatite layer.     

Fabrication of hydroxycarbonate apatite coatings with hierarchically porous structures

Hierarchically porous hydroxycarbonate apatite is known to have a high bioactivity to regenerate bone, but its application in bone graft substitutes has been restricted due to its poor mechanical properties. This drawback has been addressed by (i) depositing calcium carbonate coatings on Ti6Al4V substrates by electrophoresis; and (ii) converting the coatings to hydroxycarbonate apatite coatings with hierarchically porous structures by treatment with a phosphate buffer solution (PBS). After soaking calcium carbonate coatings in PBS for 1 day, calcium-deficient hydroxycarbonate apatite nanocrystals are deposited on the surfaces of calcium carbonate particles via a dissolution-precipitation reaction. The aggregation of the nanocrystals produces plate-like hydroxycarbonate apatite. Mesopores with a pore size of approximately 3.8nm and macropores or apertures with an aperture size of approximately 1 microm are formed within and among the plates, respectively. After soaking for 9 days, the pore size of mesopores decreases and the mesopores disappear partly due to the crystal growth of hydroxycarbonate apatite. Simulated body fluid immersion tests reveal that the good in vitro bioactivity of hydroxycarbonate apatite coatings is attributed to the calcium deficiencies in apatite lattice and the hierarchically porous structures.     

Structure and immersion behavior of plasma-sprayed apatite-matrix coatings

The microstructure and properties of a series of plasma-sprayed coatings from sinter-granulated powders fabricated from SiO2, CaO, P2O5 and Na2O-containing HA composite powders on Ti-6Al-4V substrate were reported. The immersion behavior of these coatings in a simulated body fluid (SBF) was also investigated. The results showed that sinter-granulated apatite-matrix powders were irregularly shaped and appeared quite similar. XRD patterns showed that during fabrication of the powders, P2O5 and SiO2 enhanced the decomposition of HA structure, while CaO and Na2O did not. Reasonably high bond strengths (45-50 MPa) were obtained from all coatings. The plasma spray process itself enhanced the decomposition of apatite and chemical reactions among different phases. When immersed in SBF, the intensities of such phases as alpha- and beta-TCP in all coatings decreased with immersion time and an apatite precipitation took place on all coating surfaces. The immersed SiO2- and CaO-containing HA (HSC) coating had the highest rate of apatite precipitation among all coatings. The variations in calcium ion concentration in simulated body fluid indicated that the HSC-immersed solution reached its maximal Ca concentration the earliest, while the HSCP (HA, SiO2, CaO and P2O5)-immersed solution reached its maximum the latest.     

Sol-gel hydroxyapatite coatings on stainless steel substrates.

Thin film hydroxyapatite deposits onto sandblasted 316L stainless steel substrates were prepared using water-based sol-gel technique recently developed in our lab. The coatings were annealed in air at 375 degrees C, 400 degrees C, and 500 degrees C. Phase formation, surface morphology, interfacial microstructure, and interfacial bonding strength of the coatings were investigated. Apatitic structure developed within the coatings while annealing at temperatures > or = 400 degrees C, while those heat-treated at 375 degrees C showed poor crystallinity. The coatings were dense and firmly attached to the underlying substrates, reaching an average bonding strength (as determined through the pull-out test) of 44 MPa. Nano-porous structure was found for the coatings annealed at 500 degrees C, believed to result from grain growth, and causing a slight decrease in the bonding strength. Surface microcracking, although not extensive, occurred after annealing at temperatures > or = 400 degrees C, and was linked to non-uniform thickness of the coating due to roughness of the substrate. A contraction of the coatings as a result of sintering, and phase transition from amorphous (or poor crystalline) to reasonably good crystalline apatite, may be responsible for the loss of structural integrity of the thicker sections of the coatings. It seems quite promising that a dense and adhesive apatite coating can be achieved through water-based sol gel technology after short-term annealing at around 400 degrees C in air.     

Novel silicon-doped hydroxyapatite (Si-HA) for biomedical coatings: an in vitro study using acellular simulated body fluid

Magnetron co-sputtering was used to produce silicon-doped hydroxyapatite (Si-HA) as coatings intended for potential applications such as orthopedic and dental implants. It was found that the crystallinity of the as-sputtered coatings increased after annealing, resulting in a nanocrystalline apatite structure. Subsequently, the bioactivity of the coatings was evaluated in an acellular simulated body fluid (SBF). Physicochemical evaluation demonstrated that a carbonate-containing apatite layer, which is essential for bonding at the bone/implant interface, was formed on the coating surfaces after immersion in SBF between 4 and 7 days. The annealed coatings exhibited enhanced bioactivity and chemical stability under physiological conditions, as compared with the as-sputtered coatings. It is proposed that the rate at which the carbonate-containing apatite layer forms is dependent on the scale factor of the structure. A nanocrystalline structure can provide a higher number of nucleation sites for the formation of apatite crystallites, leading to a more rapid precipitation of carbonate-containing apatite layer. This work shows that Si-HA coatings offer considerable potential for applications in hard tissue replacement, owing to their ability to form a carbonate-containing apatite layer rapidly.     

Surface characterization of Ca-P/Ag/TiO2 nanotube composite layers on Ti intended for biomedical applications

The new generation of medical implants made by titanium is functionalized with different coatings to improve their bioactivity and reduce a risk of infection. This article describes how these goals can be achieved via deposition of silver nanoparticles and calcium phosphate coating. TiO(2) nanotubes were grown on a Ti substrate via electrochemical oxidation at constant voltage in a mixture of glycerol, deionized water, and NH(4) F. Silver particles with a size of 2-50 nm were deposited on the surface using the sputter deposition technique. Calcium phosphate coatings were grown on the nanotubular titania by simple immersion in Hanks' solution. It has been found that the silver nanoparticles are distributed homogeneously in the coating, which is promising for maintaining a steady antibacterial effect. The results show also that the Ag-incorporated TiO(2) nanotubes significantly stimulate apatite deposition from Hanks' solution. The highly ordered Ag-incorporated TiO(2) nanotube arrays with apatite coating may offer unique surface features for biomedical implants, ensuring both biocompatibility and antibacterial properties.     

Fluoridated apatite coatings on titanium obtained by electron-beam deposition

In this report, a series of fluoridated apatite coatings were obtained by the electron-beam deposition method. The fluoridation of the apatite was aimed to improve the stability of the coating and elicit the fluorine effect, which is useful in the dental restoration area. Apatites fluoridated at different levels were used as initial evaporants for the coatings. The as-deposited coatings were amorphous, but after heat treatment at 500 degrees C for 1 h, the coatings crystallized well to an apatite phase without forming any cracks. The adhesion strengths of the as-deposited coatings were about 40 MPa. After heat treatment at 500 degrees C, the strengths of the pure HA and FA coatings decreased to about 20 MPa, however, the partially fluoridated coatings maintained their initial strength. The dissolution rate of the fluoridated coatings was lower than that of the pure HA coating, and the rate was the lowest in the coatings with 25% and 50% fluorine substitutions. The osteoblast-like cells responded to the coatings in a similar manner to the dissolution behavior. The cells on the fluoridated coatings showed a lower (p < 0.05) proliferation level compared to those on the pure HA coating. The alkaline phosphatase activity of the cells was slightly lower than that on the pure HA coating, but this difference was not statistically significant.     

Plasma-treated nanostructured TiO(2) surface supporting biomimetic growth of apatite

Although some types of TiO(2) powders and gel-derived films can exhibit bioactivity, plasma-sprayed TiO(2) coatings are always bioinert, thereby hampering wider applications in bone implants. We have successfully produced a bioactive nanostructured TiO(2) surface with grain size smaller than 50 nm using nanoparticle plasma spraying followed by hydrogen plasma immersion ion implantation (PIII). The hydrogen PIII nano-TiO(2) coating can induce bone-like apatite formation on its surface after immersion in a simulated body fluid. In contrast, apatite cannot form on either the as-sprayed TiO(2) surfaces (both <50 nm grain size and >50 nm grain size) or hydrogen-implanted TiO(2) with grain size larger than 50 nm. Hence, both a hydrogenated surface that gives rise to negatively charged functional groups on the surface and small grain size (<50 nm) that enhances surface adsorption are crucial to the growth of apatite. Introduction of surface bioactivity to plasma-sprayed TiO(2) coatings, which are generally recognized to have excellent biocompatibility and corrosion resistance as well as high bonding to titanium alloys, makes them more superior than many current biomedical coatings.     

Proliferation and differentiation of osteoblast-like MC3T3-E1 cells on biomimetically and electrolytically deposited calcium phosphate coatings

Biomimetic and electrolytic deposition are versatile methods to prepare calcium phosphate coatings. In this article, we compared the effects of biomimetically deposited octacalcium phosphate and carbonate apatite coatings as well as electrolytically deposited carbonate apatite coating on the proliferation and differentiation of mouse osteoblast-like MC3T3-E1 cells. It was found that MC3T3-E1 cells cultured on the biomimetically deposited carbonate apatite coating demonstrated the greatest proliferation rate and the highest differentiation potential. Cells on the biomimetically deposited octacalcium phosphate coating had lower proliferation rate before day 7, but higher after that, than those on the electrolytically deposited carbonate apatite coating. There was no difference on the expression of early differentiation markers, that is, alkaline phosphatase activity and collagen content, between biomimetically deposited octacalcium phosphate and electrolytically deposited carbonate apatite coatings. However, higher expression of late differentiation markers, that is, osteocalcin and bone sialoprotein mRNA, was found on the biomimetically deposited octacalcium phosphate coating on day 14. These results suggest that the difference in in vitro osteoblast cell performance of calcium phosphate coatings might relate to their physicochemical properties. Biomimetic carbonate apatite coating is the most favorable surface for the proliferation and differentiation of MC3T3-E1 cells.     

Strontium-substituted apatite coating grown on Ti6Al4V substrate through biomimetic synthesis

During the last few years Strontium has been shown to have beneficial effects when incorporated at certain doses in bone by stimulating bone formation. It is believed that its presence locally at the interface between an implant and bone will enhance osteointegration and therefore, ensure the longevity of a joint prosthesis. In this study we explore the possibility of incorporating Sr into nano-apatite coatings prepared by a solution-derived process according to an established biomimetic methodology for coating titanium based implants. The way this element is incorporated in the apatite structure and its effects on the stereochemistry and morphology of the resulting apatite layers was investigated, as well as its effect in the mineralization kinetics. By using the present methodology it was possible to incorporate increasing amounts of Sr in the apatite layers. Sr was found to incorporate in the apatite layer through a substitution mechanism by replacing Ca in the apatite lattice. The presence of Sr in solution induced an inhibitory effect on mineralization, leading to a decrease in the thickness of the mineral layers. The obtained Sr-substituted biomimetic coatings presented a bone-like structure similar to the one found in the human bone and therefore, are expected to enhance bone formation and osteointegration.     

Fatigue behavior of calcium phosphate coatings with different stability under dry and wet conditions.

To obtain stable plasma sprayed calcium phosphate coatings, coatings with a high crystallinity and low solubility were developed. However, stability of ceramic coatings is also influenced by their fatigue resistance. Recently, fatigue failure was proposed to explain coating detachment from implants under loaded conditions. Therefore, plasma-sprayed calcium phosphate coatings with different crystallinity were investigated in vitro for fatigue failure. An amorphous and a crystalline hydroxylapatite coating (AHA and CHA) and a highly crystalline fluorapatite coating (FA) were subjected to cyclic load tests, both in dry conditions and in simulated body fluid (SBF). The results in SBF revealed that the crystalline CHA and FA coating detached completely at the highest stressed middle section of the bar. The FA coating delaminated earlier than the CHA coating. The amorphous AHA coating showed only partial coating loss at the completion of the test. Tests in dry conditions did not reveal any change in the coatings tested. These results suggest a relation between crystallinity of apatite coatings and their failure due to fatigue: high crystallinity coatings demonstrate earlier and more complete fatigue failure than the amorphous apatite coatings. It can be concluded that coating stability is not determined solely by static dissolution, but by fatigue failure as well.     

Silicon-substituted hydroxyapatite thin films: effect of annealing temperature on coating stability and bioactivity

The effect of annealing temperature on the physicochemical and biological characteristics of magnetron cosputtered silicon-substituted hydroxyapatite (SiHA) thin coatings was studied. Annealing is required to transform as-sputtered amorphous films into crystalline coatings. A nanocrystalline, single-phase apatite structure was achieved for coatings heated to 600 or 700 degrees C and, with increasing annealing temperature, the crystallite size increased. Small crystallites were found to be more soluble in the physiological environment but, at the same time, were able to induce early formation of a new apatite layer. A human osteoblast-like (HOB) cell model was used to evaluate the performance of these annealed SiHA coatings. HOB cells attached and grew well on coatings and, after 42 days in culture, a mineralization process was observed to be taking place, with evidence of calcium phosphate minerals throughout the extracellular matrix. Our findings indicated that an annealing temperature of 600 degrees C is sufficient to achieve crystalline SiHA coatings and exhibiting good chemical stability and bioactivity.     

Fabrication and in vitro characterization of magnetic hydroxycarbonate apatite coatings with hierarchically porous structures

Hydroxycarbonate apatite/Fe(3)O(4) composite coatings (MHACs) with hierarchically porous structures were fabricated by electrophoretic deposition of CaCO(3)/Fe(3)O(4) particles on Ti6Al4V substrates followed by treatment with phosphate buffer solution (PBS) at 37 degrees C. The effects of Fe(3)O(4) on the conversion rate of calcium carbonate to hydroxycarbonate apatite and the porous structures and in vitro bioactivity of MHACs were investigated. After soaking CaCO(3)/Fe(3)O(4) coatings in PBS, hydroxycarbonate apatite nucleates heterogeneously on the surfaces of CaCO(3)/Fe(3)O(4) particles and forms a plate-like structure. Fe(3)O(4) increases the velocity of nucleus formation of hydroxycarbonate apatite. After soaking for 1day, the percentage of unreacted calcium carbonate for MHACs is approximately 9.1%, lower than the approximately 41.0% for hydroxycarbonate apatite coatings (HCACs). As the CaCO(3)/Fe(3)O(4) coatings are converted to MHACs, macropores with a pore size of approximately 4mum on the coatings and mesopores with a pore size of approximately 3.9nm within the hydroxycarbonate apatite plates are formed. The mesopores remain in the MHACs after treatment with PBS for 9 days, while they disappear in the HCACs. Simulated body fluid immersion tests reveal that Fe(3)O(4) improves the in vitro bioactivity of biocoatings. The amount of bone-like apatite precipitated on the surfaces of MHACs is greater than that on the surfaces of HCACs.     

Osteoclastic resorption of biomimetic calcium phosphate coatings in vitro

A new biomimetic method for coating metal implants enables the fast formation of dense and homogeneous calcium phosphate coatings. Titanium alloy (Ti6Al4V) disks were coated with a thin, carbonated, amorphous calcium phosphate (ACP) by immersion in a saturated solution of calcium, phosphate, magnesium, and carbonate. The ACP-coated disks then were processed further by incubation in calcium phosphate solutions to produce either crystalline carbonated apatite (CA) or octacalcium phosphate (OCP). The resorption behavior of these three biomimetic coatings was studied using osteoclast-enriched mouse bone-marrow cell cultures for 7 days. Cell-mediated degradation was observed for both carbonated apatite and octacalcium phosphate coatings. Numerous resorption lacunae characteristic of osteoclastic resorption were found on carbonated apatite after cell culture. The results showed that carbonated apatite coatings are resorbed by osteoclasts in a manner consistent with normal osteoclastic resorption. Osteoclasts also degraded the octacalcium phosphate coatings but not by classical pit formation.     

Fluor-hydroxyapatite sol-gel coating on titanium substrate for hard tissue implants

Hydroxyapatite (HA) and fluor-hydroxyapatite (FHA) films were deposited on a titanium substrate using a sol-gel technique. Different concentrations of F- were incorporated into the apatite structure during the sol preparation. Typical apatite structures were obtained for all coatings after dipping and subsequent heat treatment at 500 degrees C. The films obtained were uniform and dense, with a thickness of approximately 5 microm. The dissolution rate of the coating layer decreased with increasing F- incorporation within the apatite structure, which demonstrates the possibility of tailoring the solubility by a functional gradient coating of HA and FHA. The cell proliferation rate on the coating layer decreased slightly with increasing F- incorporation. The alkaline phosphatase (ALP) activity of the cells on all the HA and FHA coated samples showed much higher expression levels compared to pure Ti. This confirmed the improved activity of cell functions on the substrates with the sol-gel coating treatment.     

Mechanism and kinetics of apatite formation on nanocrystalline TiO2 coatings: a quartz crystal microbalance study

Apatite (Ca5(PO4)3OH) has long been considered as an excellent biomaterial to promote bone repairs and implant. Apatite formation induced by negatively charged nanocrystalline TiO2 coatings soaked in simulated body fluid (SBF) was investigated using in situ quartz crystal microbalance (QCM), scanning electron microscopy (SEM), Fourier-transformed infrared spectroscopy (FTIR), X-ray diffraction (XRD) and energy-dispersive X-ray spectroscopy (EDX) techniques, and factors affecting its formation such as pH, size of TiO2 particles and thickness of TiO2 coatings, were discussed in detail. Two different stages were clearly observed in the process of apatite precipitation, indicating two different kinetic processes. At the first stage, the calcium ions in SBF were initially attracted to the negatively charged TiO2 surface, and then the calcium titanate formed at the interface combined with phosphate ions, consequently forming apatite nuclei. After the nucleation, the calcium ions, phosphate ions and other minor ions (i.e. CO3(2-) and Mg2+) in supersaturated SBF deposited spontaneously on the original apatite coatings to form apatite precipitates. In terms of the in situ frequency shifts, the growth-rate constants of apatite (K1 and K2) were estimated, respectively, at two different stages, and the results were (1.96+/-0.14)x10(-3)s(-1) and (1.28+/-0.10)x10(-4)s(-1), respectively, in 1.5 SBF solution. It was found that the reaction rate at the first stage is obviously higher than that at the second stage.     

Adherent apatite coating on titanium substrate using chemical deposition

Plasma-sprayed "HA" coatings on commercial orthopedic and dental implants consist of mixtures of calcium phosphate phases, predominantly a crystalline calcium phosphate phase, hydroxyapatite (HA) and an amorphous calcium phosphate (ACP) with varying HA/ACP ratios. Alternatives to the plasma-spray method are being explored because of some of its disadvantages. The purpose of this study was to deposit an adherent apatite coating on titanium substrate using a two-step method. First, titanium substrates were immersed in acidic solution of calcium phosphate resulting in the deposition of a monetite (CaHPO4) coating. Second, the monetite crystals were transformed to apatite by hydrolysis in NaOH solution. Composition and morphology of the initial and final coatings were identified using X-ray diffraction (XRD), Scanning Electron Microscopy, and Energy Dispersive Spectroscopy (EDS). The final coating was porous and the apatite crystals were agglomerated and followed the outline of the large monetite crystals. EDS revealed the presence of calcium and phosphorous elements on the titanium substrate after removing the coating using tensile or scratching tests. The average tensile bond of the coating was 5.2 MPa and cohesion failures were observed more frequently than adhesion failures. The coating adhesion measured using scratch test with a 200-microm-radius stylus was 13.1N. Images from the scratch tracks demonstrated that the coating materials were squashed without fracturing inside and/or at the border of the tracks until the failure point of the coating. In conclusion, this study showed the potential of a chemical deposition method for depositing a coating consisting of either monetite or apatite. This method has the advantage of producing a coating with homogenous composition on even implants of complex geometry or porosity. This method involves low temperatures and, therefore, can allow the incorporation of growth factors or biogenic molecules.     

Biomineralized strontium-substituted apatite/titanium dioxide coating on titanium surfaces

Bone mineral is a multi-substituted calcium phosphate. One of these ion substitutions, strontium, has been proven to increase bone strength and decrease bone resorption. Biomimetics is a potential way to prepare surfaces that provide a favorable bone tissue response, thus enhancing the fixation between bone and implants. Here we prepared double-layered strontium-substituted apatite and titanium dioxide coatings on titanium substrates via mimicking bone mineralization. Morphology, crystallinity, surface chemistry and composition of Sr-substituted coatings formed via biomimetic coating deposition on crystalline titanium oxide substrates were studied as functions of soaking temperature and time in phosphate buffer solutions with different Sr ion concentration. The morphology of the biomimetic apatite changed from plate-like for the pure HA to sphere-like for the Sr ion substituted. Surface analysis results showed that 10–33% of Ca ions in the apatite have been substituted by Sr ions, and that the Sr ions were chemically bonded with apatite and successfully incorporated into the structure of apatite.     

Biomimetic deposition of apatite coating on surface-modified NiTi alloy

TiO(2) coatings were prepared on NiTi alloy by heat treatment in air at 300, 400, 600 and 800 degrees C. The heat-treated NiTi alloy was subsequently immersed in a simulated body fluid for the biomimetic deposition of the apatite layer onto the surface of TiO(2) coating. The apatite coatings as well as the surface oxide layer on NiTi alloy were characterized using scanning electron microscopy equipped with energy dispersive spectroscopy, X-ray diffraction, X-ray photoelectron spectroscopy and Raman spectroscopy. Results showed the samples heat-treated at 600 degrees C produced a layer of anatase and rutile TiO(2) on the surface of NiTi. No TiO(2) was detected on the surface of NiTi after heat treatment at 300 and 400 degrees C by X-ray diffraction, while rutile was formed on the surface of the 800 degrees C heat-treated sample. It was found that the 600 degrees C heat-treated NiTi induced a layer consisted of microcrystalline carbonate containing hydroxyapatite on its surface most effectively, while 300 and 400 degrees C heat-treated NiTi did not form apatite. This was due to the presence of anatase and/or rutile in the 600 and 800 degrees C heat-treated NiTi which could provide atomic arrangements in their crystal structures suitable for the epitaxy of apatite crystals, and anatase had better apatite-forming ability than rutile. XPS and Raman results revealed that this apatite layer was a carbonated and non-stoichiometric apatite with Ca/P ratio of 1.53, which was similar to the human bone. The formation of apatite on 600 degrees C heat-treated NiTi following immersion in SBF for 3 days indicated that the surface modified NiTi possessed excellent bioactivity.