Formation and bioactivity of porous and nanostructured TiO2/beta-TCP coating on titanium.

Titanium and its alloys have been widely used as hard tissue implants due to their excellent mechanical properties and biocompatibility. However, their near bio-inertness and metallic ion release are still the problems with clinical uses. In this paper, porous and nanostructured TiO2/beta-tricalcium phosphate (beta-TCP) composite coatings were prepared on titanium substrates by plasma electrolytic oxidation (PEO) in a Ca and P-containing electrolyte. The influence of PEO electric current density on phase composition and bioactivity of the coatings were studied. X-ray diffraction, scanning electron microscopy and Fourier transfer infrared spectroscopy were utilized to characterize the phase composition and microstructure of the coatings. Simulated body fluid immersion tests were employed on the coatings to evaluate their bioactivity. The results reveal that TiO2/beta-TCP composite coating with pores size less than 10 microm and grains of 50-100 nm in size was prepared. The electric current density of PEO is an important factor in the formation of the composite coating. The TiO2/beta-TCP composite coating shows good bioactivity, which are attributed to the incorporation of beta-TCP.

Nano-structured titanium coating for improving biological performance.

Nano-structured titanium coating was obtained by alkali treating the vacuum plasma sprayed samples following hot water immersing for 24 h. The influences of the surface microstructure on the biological performance were studied. A canine model was applied for in vivo evaluation of the bone bonding ability of the coatings. The histological examination results demonstrate that new bone was formed more rapidly on the nano-structured coating implants and grew into the porosity than the as-sprayed one. After 4 weeks implantation, the nano-structured implants were found to appose directly to the surrounding bone while large lacunae could still be observed at the interface between the as-sprayed samples and bone. All these results indicate that a nano-structured surface on the porous titanium coating is favorable for bone bonding.

UV-irradiation-induced bioactivity on TiO2 coatings with nanostructural surface.

Titania (TiO2) coatings with nanostructural surface prepared using plasma spraying technology were irradiated by ultraviolet light in simulated body fluids to improve their bioactivity. The in vitro bioactivity of the coatings was evaluated by investigating the formation of apatite on their surfaces in simulated body fluids. Bone-like apatite was observed to precipitate on the UV-irradiated TiO2 coating with nanostructural surface after it was immersed in simulated body fluid for a certain period, but not on the as-sprayed and UV-irradiated TiO2 coatings without nanostructural surface. The results indicate that the nano-TiO2 surface can be activated by UV-irradiation to induce its bioactivity. The ability of apatite formation on the nano-TiO2 surface was improved with the increase of UV-irradiation time. The in vivo results reveal that the as-prepared TiO2 coating with nanostructural surface cannot induce the formation of new bones during the implantation period, but the UV-irradiated TiO2 coating with nanostructural surface could do so during an implantation time longer than 2 months. Our results indicate that the osseointegration ability of the plasma-sprayed TiO2 coating with nanostructural surface can be improved by UV irradiation.

Hydrogen plasma surface activation of silicon for biomedical applications.

Silicon has gradually been recognized to be an essential trace element in the normal metabolism of higher animals, and the role of silicon in the human body has aroused interests in the biomedical community. In fact, the interactions between silicon-based devices and the human body such as biosensors and microelectromechanical systems (MEMS) often suffer from poor biocompatibility. In this work, hydrogen plasma immersion ion implantation (H-PIII) is conducted to improve the bioactivity or bone conductivity of silicon. In order to investigate the formation mechanism of bone-like apatite on the surface of the hydrogen implanted silicon wafer, two comparative experiments, hydrogenation and argon bombardment, are performed. The H-PIII sample exhibits an amorphous surface consisting of Si-H bonds. After immersion in simulated body fluids, a negatively charged surface containing the functional group ([triple bond]Si-O-) is produced and bone-like apatite is observed to nucleate and grow on the surface. The surface of the H-PIII silicon wafer favors the adhesion and growth of osteoblast cells and good cytocompatibility may be inferred.

Bioactivity and cytocompatibility of silicon wafers treated by water.

A range of bioactive ceramics can induce bone-like apatite to deposit on their surface in simulated body fluid (SBF). In this work, the silicon wafer was treated using deionized water to improve its bioactivity. The morphology and chemical composition of the treated silicon wafer was examined using Fourier transform infrared spectroscopy, atomic force microscopy and X-ray photoelectron spectroscopy. Field emission scanning electron microscopy was used to observe the surface morphologies of silicon wafers soaked in SBF. The results indicated that a hydrated sub-oxide film having Si--OH groups formed on the surface of the silicon wafer after the water treatment. The amount of Si--OH groups increased with raising the treatment temperature or prolonging the immersion time. Apatite was deposited on the surface of water-treated silicon wafers immersed in SBF. The apatite deposition was correlated with the amount of Si--OH groups. Human mesenchymal stem cells cultured on the surface of the water-treated silicon wafers showed good adhesion and spread, indicating that the cytocompatibility of silicon wafer was enhanced by this water treatment.

Biocompatibility and antibacterial activity of plasma sprayed titania coating grafting collagen and gentamicin.

In this article, the plasma sprayed titania coatings were treated by grafting pure and gentamicin loaded collagen to improve the biocompatibility and antibacterial activity. The biocompatibility of the titania coating grafting collagen was evaluated by in vitro cell culturing test. The release rate of gentamicin from collagen was measured in tris-HCl buffer using UV spectrophotometer, and the antibacterial activity of the titania coating with gentamicin against Staphylococcus aureus was examined using the zone of inhibition test. The results showed that collagen was successfully grafted on the surface of titania coatings treated by sulfuric acid. The in vitro cell culturing test revealed that collagen significantly improved the cell adhesion and proliferation on the surface of titania coatings. The gentamicin loaded in collagen matrix could retain a sustained release in tris-HCl for 30 days, which was efficient to protect against the postoperative infection caused by S. aureus. The results indicated that the plasma sprayed titania coating grafting collagen and gentamicin would have the antibacterial activity together with the biocompatibility, which might be beneficial for the long term stability and surgical success rate of implants.

Bioactivity and cytocompatibility of zirconia (ZrO(2)) films fabricated by cathodic arc deposition.

Zirconium oxide thin films were fabricated on silicon wafers using a filtered cathodic arc system in concert with oxygen plasma. The structure and phase composition of the zirconium oxide thin films were characterized by atomic force microscopy (AFM), X-ray diffraction (XRD), Rutherford backscattering spectrometry (RBS), and transmission electron microscopy (TEM). The bioactivity was assessed by investigating the formation of apatite on the film surface after soaking in simulated body fluids. Bone marrow mesenchymal stem cells (BMMSC) were used to further evaluate the cytocompatibility of the materials. The results indicate that the films are composed of stoichiometric ZrO(2) and the composition is quite uniform throughout the thickness. Bone-like apatite can be formed on the surface of the ZrO(2) thin film in our SBF immersion experiments, suggesting that the surface is bioactive. The outermost layer of the ZrO(2) thin film comprises nano-sized particles that can be identified by AFM images taken on the thin film surface and TEM micrographs obtained from the interface between the ZrO(2) thin film and apatite layer. The nanostructured surface is believed to be the key factor that apatite is induced to precipitate on the surface. Bone marrow mesenchymal stem cells are observed to grow and proliferate in good states on the film surface. Our results show that ZrO(2) thin films fabricated by cathodic arc deposition exhibit favorable bioactivity and cytocompatibility.

In vivo evaluation of plasma-sprayed wollastonite coating.

Wollastonite coatings were prepared by plasma spraying. The bioactivity of wollastonite coatings was investigated in vivo by implanting in dog's muscle, cortical bone and marrow, respectively. The behaviour of bone tissue around wollastonite coatings were examined by histological and SEM observation. After 1 month in the muscle, a bone-like apatite layer was found to form on the surface of the wollastonite coating. When implanted in cortical bone, histological observation demonstrated that bone tissue could extend and grow along the surface of the wollastonite coating. The coating bonded directly to the bone without any fibrous tissue, indicating good biocompatibility and bone conductivity. SEM and EDS analysis revealed that bone did not bond to wollastonite coating directly, but through a Ca/P layer. This suggested that the formation of bone-like apatite layer was very important for bonding to the bone tissue. The amount of bone-implant contact was also measured. Wollastonite coating was shown to stimulate more bone formation on its surface than titanium coating after implantation for 1 month, enhancing the short-term osseointegration properties of implant. The test in marrow indicated that wollastonite coatings could induce new bone formation on their surface showing good bone inductivity property.

In vivo evaluation of plasma-sprayed titanium coating after alkali modification.

In this paper, plasma-sprayed titanium coatings were modified by alkali treatment. The changes in chemical composition and structure of coatings were examined by SEM and AES. The results obtained indicated that a net-like microscopic texture feature, which was full of the interconnected fine porosity, appeared on the surface of alkali-modified titanium coatings. The surface chemical composition was also altered by alkali modification. A sodium titanate compound was formed on the surface of the titanium coating and replaced the native passivating oxide layer. Its thickness was measured as about 150 nm which was about 10 times of that of the as-sprayed coating. The bone bonding ability of titanium coatings were investigated using a canine model. The histological examination and SEM observation demonstrated that more new bone was formed on the surface of alkali-modified implants and grew more rapidly into the porosity. The alkali-modified implants were found to appose directly to the surrounding bone. In contrast, a gap was observed at the interface between the as-sprayed implants and bone. The push-out test showed that alkali-modified implants had a higher shear strength than as-sprayed implants after 1 month of implantation. An interfacial layer, containing Ti, Ca and P, was found to form at the interface between bone and the alkali-modified implant by EDS analysis.

Improvement of surface bioactivity on titanium by water and hydrogen plasma immersion ion implantation.

We have investigated the surface bioactivity of titanium after water and hydrogen plasma immersion ion implantation. Plasma immersion ion implantation (PIII) excels in the surface treatment of components possessing a complicated shape such as medical implants. In addition, water and hydrogen PIII has been extensively studied as a method to fabricate silicon-on-insulator (SOI) substrates in the semiconductor industry and so it is relatively straightforward to transfer the technology to the biomedical field. In our investigation, water and hydrogen were plasma-implanted into titanium sequentially. Our objective is that water PIII introduces near-surface damages that trap hydrogen implanted in the subsequent step to improve the surface bioactivity while the desirable bulk properties of the materials are not compromised. Ti-OH functional groups can be detected on the (H(2)O+H(2))-implanted titanium surface by X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FTIR) spectroscopy. After incubation in simulated body fluids (SBF) for cytocompatibililty evaluation in vitro, bone-like hydroxyapatite was found to precipitate on the (H(2)O+H(2)) implanted samples while no apatite was found on titanium samples plasma implanted with water or hydrogen alone. Human osteoblast cells were cultured on the (H(2)O+H(2))-implanted titanium surface and they exhibited good adhesion and growth. Our results suggest a practical means to improve the surface bioactivity and cytocompatibility of medical implants made of titanium.

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.

Acid-induced bioactive titania surface.

Nano- and conventional-TiO(2) powders were deposited onto titanium alloy using atmospheric plasma spraying technology. As-sprayed titania coatings were treated by H(2)SO(4) and HCl solutions at room temperature for 24 h, and the bioactivity was evaluated by simulated body fluid tests. Measured X-ray diffraction patterns indicated that as-sprayed titania coatings obtained from both nano and conventional powders were composed of primary rutile as well as a small quantity of anatase and Ti(3)O(5). The surface of as-sprayed coatings prepared from the conventional powder was rougher than that from nanopowder. After immersion in simulated body fluid for 2 weeks, both acid-treated nano- and titania coatings have induced carbonate-containing hydroxyapatite to form on the surfaces. However, this phenomenon did not appear on the surface of as-sprayed nano- and conventional-titania coatings. The results obtained indicated that the bioactivity of plasma sprayed titania coatings was improved by the acid treatment and had nothing to do with phase composition and particle size of the original TiO(2) powders.

Effect of hydroxyapatite coating crystallinity on dissolution and osseointegration in vivo.

Hydroxyapatite (HA) coating with different crystallinities were deposited by plasma spraying and vapor-flame treatment process. Their crystallinities are about 55 and 98%, respectively. These coatings were implanted in cortical bone, muscle, and marrow of dogs. The dissolution and osseointegration behavior were evaluated by scanning electron microscope (SEM) observation histological analysis. The results obtained indicated that after implanted in muscle, a bone-like apatite layer was formed on the surface of as-sprayed coating, which was not observed on the surface of the treated coating. The as-sprayed coating has the ability to induce new bone formation on its surface after implanted in marrow. In contrast, the treated coating displays a limited bone bioactivity. The vapor-flame process diminishes the short-term osseointegration properties of the HA coating, but no significant affection was found after three months implantation. Either in muscle or in cortical bone, treated coating exhibits higher stability than the as-sprayed coating, in some conditions.

Early apatite deposition and osteoblast growth on plasma-sprayed dicalcium silicate coating.

Dicalcium silicate coating was deposited onto a Ti-6Al-4V substrate using plasma-spraying technology. The coating was immersed in simulated body fluid (SBF) for 1, 3, 6, 12, 24, and 48 h to investigate early apatite formation on the coating. Osteoblasts were also seeded onto the surface of the dicalcium silicate coating to evaluate its biocompatibility. Cold field-emission scanning electron microscopy and energy-dispersive X-ray spectrometry were used to evaluate the morphologies and determine the chemical composition of the coatings. The surface structural changes caused by immersion in SBF were analyzed using thin-film X-ray diffraction. After the dicalcium silicate coating was soaked in SBF solution 1-6 h, two types of particles containing calcium and phosphorus were formed on the surface. One type consisted of relatively larger particles (P1) precipitated on the surface of the coating from the precursor cluster formed in the SBF solution. The second type was composed of particles (P2) nucleated on the surface of the coating. With increasing immersion time, the particles coalesced to form a surface Ca-P layer. The Ca-P layer was composed of amorphous calcium phosphate that was not transformed to crystalline apatite until the immersion time in SBF exceeded 24 h. The formation mechanism of the Ca-P layer and apatite on the surface of the coating is believed to be involved in the formation of the Si 3-ring active surface site with negative charge. The cell-seeding test revealed that osteoblasts grew and proliferated very well on the surface of the dicalcium silicate coating.

In vitro degradation behavior and cytocompatibility of biodegradable AZ31 alloy with PEO/HT composite coating.

Biodegradable magnesium-based implants have attracted much attention recently in orthopedic applications because of their good mechanical properties and biocompatibility. However, their rapid degradation in vivo will not only reduce their mechanical strength, but also induce some side effects, such as local alkalization and gas cavity, which may lead to a failure of the implant. In this work, a hydroxyapatite (HA) layer was prepared on plasma electrolytic oxidization (PEO) coating by hydrothermal treatment (HT) to fabricate a PEO/HT composite coating on biodegradable AZ31 alloy. The in vitro degradation behaviors of all samples were evaluated in simulated body fluid (SBF) and their surface cytocompatibility was also investigated by evaluating the adhesion and proliferation of osteoblast cells (MC3T3-E1). The results showed that the HA layer consisted of a dense inner layer and a needle-like outer layer, which successfully sealed the PEO coating. The in vitro degradation tests showed that the PEO/HT composite coating improved the corrosion resistance of AZ31 alloy in SBF, presenting nearly no severe local alkalization and hydrogen evolution. The lasting corrosion resistance of the PEO/HT composite coating may attribute to the new hydroxyapatite formation during the degradation process. Moreover, compared with AZ31 alloy and PEO coating, PEO/HT composite coating was more suitable for cells adhesion and proliferation, indicating improved surface cytocompatibility. The results show that the PEO/HT composite coating is promising as protective coating on biodegradable magnesium-based implants to enhance their corrosion resistance as well as improve their surface cytocompatibility for orthopedic applications.

Effects of graphene plates' adoption on the microstructure, mechanical properties, and in vivo biocompatibility of calcium silicate coating.

Calcium silicate (CS) ceramic is a good coating candidate for biomedical implants to improve biocompatibility and accelerate early osseointegration. However, the poor fracture toughness and wear resistance of this ceramic material restricts the long-term performance of implants. In this study, graphene plates (GPs) were used as reinforcement to improve the mechanical properties of CS coating. Composite coating containing 1.5 weight % GPs was prepared by vacuum plasma spraying technology. The good survival of the GPs in the composite coating was demonstrated by Raman analysis, although the defects of the GPs were increased after plasma spraying. Effects of the GPs' adoption on the microstructure of the coating were studied by scanning electron microscopy and transmission electron microscopy. Results showed that the GPs were homogenously distributed in the CS grains interface or enwrapped on the particles, and exhibited good wetting behavior with the CS matrix. The wear properties of the composite coating were obviously enhanced by the reinforcement of GPs. The reinforcement mechanism was attributed to the enhanced micro-hardness and interfacial bonding of the particles in the coating. In vivo experiments demonstrated that the composite coating possessed similarly good biocompatibility compared to pure CS coating. The bone-implant contact ratio reached 84.3%±7.4% for GPs/CS coating and 79.6%±9.4% for CS coating after 3 months' implantation.

Plasma sprayed wollastonite/TiO2 composite coatings on titanium alloys.

Wollastonite/TiO2 composite coatings were prepared using plasma spraying technology onto Ti-6Al-4V substrate. The composite coatings exhibit obvious lamellar structure with alternating wollastonite coating and TiO2 coating. No obvious cracks exist on the interface between coatings and substrate. In the case of composite coatings, the primarily crystalline phases of the coatings are wollastonite and rutile, indicating wollastonite and TiO2 did not react during plasma spraying process. Some of rutile in the powders transforms into anatase due to plasma spraying. The mean bond strength of the composite coatings is higher than 30 MPa. The Vickers microhardness of coatings increase with the increase in the content of TiO2. Wollastonite/TiO2 composite coatings were soaked in simulated body fluid to examine their bioactivity. Carbonate-containing hydroxyapatite (CHA) layer was formed on the surface of the wollastonite and W7T3 coatings soaked in simulated body fluid, while was not formed on the surface of the TiO2 and W3T7 coatings after immersion. In addition, a rich-silica layer appeared at the interface of CHA and wollastonite and W7T3 coatings. In order to investigate the cytocompatibility of the coatings, osteoblast was seeded onto the surface of the coatings. The scanning electron microscopy observation showed that the addition of wollastonite promote the proliferation of osteoblast. It is enough to prove that the wollastonite and wollastonite/TiO2 composite coatings possess excellent cytocompatibility.

Bioactivity of plasma sprayed dicalcium silicate coatings.

Dicalcium silicate coatings on titanium alloys substrates were prepared by plasma spraying and immersed in simulated body fluids for a period of time to investigate the nucleation and growth of apatite on the surface of the coatings. Surface structural changes of the specimens were analyzed by XRD and IR technologies. SEM and EDS were used to observe surface morphologies and determine the composition of dicalcium silicate coatings before and after immersion in simulated body fluid. The plasma sprayed dicalcium silicate coating was bonding tightly to the substrate. The coating was mainly composed of beta-Ca2SiO4 and glassy phase. A dense carbonate-containing hydroxyapatite (CHA) layer was formed on the surface of the plasma sprayed dicalcium silicate coating soaked in SBF solution for 2 days. In addition, a silica-rich layer was also observed between CHA layer and coatings. With an increase in the immersion time, the CHA layer gradually became thicker. The results obtained indicated that the plasma sprayed dicalcium silicate coating possesses excellent bioactivity.

Mechanism of apatite formation on wollastonite coatings in simulated body fluids.

The formation mechanism of apatite on the surface of wollastonite coating was examined. Plasma-sprayed wollastonite coatings were soaked in a lactic acid solution (pH=2.4) to result in the dissolution of calcium from the coating to form silanol (triple bond Si-OH) on the surface. Some calcium-drained samples were soaked in a trimethanol aminomethane solution (pH=10) for 24h to create a negatively charged surface with the functional group (triple bond Si-O(-)). These samples before and after treatment in a trimethanol aminomethane solution were immersed in simulated body fluids (SBF) to investigate the precipitation of apatite on the coating surface. The results indicate that the increase of calcium in the SBF solution is not the critical factor affecting the precipitation of apatite on the surface of the wollastonite coating and the apatite can only form on a negatively charged surface with the functional group (triple bond Si-O(-)). The mechanism of apatite formation on the wollastonite coating is proposed. After the wollastonite coatings are immersed into the SBF, calcium ions initially exchange with H(+) leading to the formation of silanol (triple bond Si-OH) on the surface of the layer and increase in the pH value at the coating-SBF interface. Consequently, a negatively charged surface with the functional group (triple bond Si-O(-)) forms on the surface. Due to the negatively charged surface, Ca(2+) ions in the SBF solution are attracted to the interface between the coating and solution, thereby increasing the ionic activity of the apatite at the interface to the extent that apatite precipitates on the coating surface.

In vivo evaluation of plasma sprayed hydroxyapatite coatings having different crystallinity.

In this paper, hydroxyapatite (HA) coatings having the crystallinities of 56% and 98% were deposited by the plasma spraying and vapor-flame treatment process. The phase composition and crystallinity of the coatings were investigated by X-ray diffraction and infrared spectra. The dissolution behavior of the coatings in tris-buffer solutions was examined. The results obtained indicated that the coating having the high crystallinity showed the lower dissolution as compared to the low crystallinity coating. The bone bonding ability of HA coatings were observed in vivo by implanted in dog's femur. After 3 months implantation, the high crystallinity coating showed the higher shear strengths and remained integrated, whereas the separation of the coating fragments was clearly observed in the coating having low crystallinity.