Effects of surface treatment of Ti-6Al-4V titanium alloy on biocompatibility in cultured human umbilical vein endothelial cells

Among the titanium alloys employed as implant materials, the Ti-6Al-4V alloy is still widely used. Ti-6Al-4V titanium alloy samples, in untreated state and subjected to treatments in air by furnace or glow-discharge processes, were put in contact with human umbilical vein endothelial cells (HUVEC) in order to evaluate their effects on biocompatibility. In HUVEC kept for 48 h in the presence of the three sample types neither cell proliferation nor protein content nor lactate dehydrogenase release in the culture medium are affected, while apoptosis is induced after 48- and 96-h contact of the cells with the untreated sample type, and after 96-h contact with the plasma treated one, the furnace treated sample type being ineffective. The expression of two adhesion molecules, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) was also studied. The incubation of HUVEC with the three sample types for 48 or 96 h induces a significant increase in ICAM-1 protein levels, in comparison with control cells, while VCAM-1 expression is not detectable. In the same way, TNF-alpha release in the culture medium, assayed after 48- and 96-h contact of the cells with the three sample types, is significantly higher, in comparison with control, even if the highest values are registered in the presence of the untreated samples. Taken together, these data indicate that, although Ti-6Al-4V alloy samples, and in particular the treated ones, show a good biocompatibility, attention must be given to the first signs of inflammation.     

Fatigue endurance of Ti-6Al-4V alloy with electro-eroded surface for improved bone in-growth

Ti-6Al-4V hour-glass shaped rotating beam specimens with duplex microstructure were processed by electric discharge machining (EDM). A comparatively high peak current of 29A was utilized in order to increase surface roughness for improved osteointegration. High cycle fatigue (HCF) tests were performed in rotating beam loading (R=-1) on these EDM specimens and results were compared with electrolytically polished specimens serving as reference. As expected, the HCF performance of EDM specimens was inferior to the electrolytically polished specimens. A detailed study of fatigue crack nucleation and microcrack growth was carried out on failed specimens by SEM. The poor HCF strength of EDM specimens is explained by early crack nucleation due to the high notch sensitivity of Ti-6Al-4V. In addition, process-induced residual tensile stresses and microstructural effects may also account for the drastic loss in HCF performance relative to the electropolished baseline.     

In vitro interaction between surface-treated Ti-6Al-4V titanium alloy and human peripheral blood mononuclear cells

The Ti-6Al-4V titanium alloy is widely employed as an implant material. The effects of Ti-6Al-4V samples, tested in both an untreated state and one in which the samples were subjected to a glow-discharge treatment performed with the use of air, on human peripheral blood mononuclear cells (PBMC) were studied. Apoptosis, undetectable after 24-h contact of PBMC with the two sample types, is induced after 48 h by treated samples, and, after 48 h, but in the presence of 1.5 microg/mL PHA, by both sample types. The expression of intercellular adhesion molecule-1 (ICAM-1) always increases, in comparison with control, in PBMC put in contact with the two sample types. In the same way, a remarkable increase in tumor necrosis-alpha (TNF-alpha) release in the culture medium is registered, when PBMC are put in contact with the two sample types for 24 and 48 h. Human umbilical-vein endothelial cells (HUVEC) cocultured for 48 h with PBMC, previously incubated with the two sample types for 24 h, show an increase in ICAM-1 and vascular cell adhesion molecule-1 (VCAM-1) protein expression in comparison with control (HUVEC cocultured with control PBMC), indicating that inflammatory phenomena might occur. Taken together, these results suggest that, although plasma-treated titanium alloy shows a better biocompatibility in comparison with the untreated one, attention must be paid to the careful control of the first signs of inflammation.     

The machinability of cast titanium and Ti-6Al-4V

This study investigated the machinability (ease of metal removal) of commercially pure (CP) titanium and Ti-6Al-4V alloy. Both CP Ti and Ti-6Al-4V were cast into magnesia molds. Two types of specimens (with alpha-case and without alpha-case) were made for CP Ti and Ti-6Al-4V. Machinability (n = 5) was evaluated as volume loss (mm3) by cutting/grinding the 3.0 mm surface using fissure burs and silicon carbide (SiC) under two machining conditions: (1) two machining forces (100 or 300 gf) at two rotational speeds (15000 or 30000 rpm) for 1 min, and (2) constant machining force of 100 gf and rotational speed of 15000 rpm for 1, 2, 5, 10, and 30 min. As controls, conventionally cast Co-Cr and Type IV gold alloys were evaluated in the same manner as the titanium. When fissure burs were used, there was a significant difference in the machinability between CP titanium with alpha-case and without alpha-case. On the other hand, there was no appreciable difference in the amount of metal removed for each tested metal when using the SiC points.     

The effect of surface treatments on the fretting behavior of Ti-6Al-4V alloy

Stem modularity in total hip replacement introduces an additional taper joint between Ti-6Al-4V stem components with the potential for fretting corrosion processes. One possible way to reduce the susceptibility of the Ti-6Al-4V/Ti-6Al-4V interface to fretting is the surface modification of the Ti-6Al-4V alloy. Among the tested, industrially available surface treatments, a combination of two deep anodic spark deposition treatments followed by barrel polishing resulted in a four times lower material release with respect to untreated, machined fretting pad surfaces. The fretting release has been quantified by means of radiotracers introduced in the alloy surface by proton irradiation. In a simple sphere on flat geometry, the semispherical fretting pads were pressed against flat, dog-bone shaped Ti-6Al-4V fatigue samples cyclically loaded at 4 Hz. In this way a cyclic displacement amplitude along the surfaces of 20 mum has been achieved. A further simplification consisted in the use of deionized water as lubricant. A comparison of the radiotracer results with an electrochemical material characterization after selected treatments by potentiostatic tests of modular stems in 0.9% NaCl at 40 degrees C for 10 days confirmed the benefit of deep anodic spark deposition and subsequent barrel polishing for improving the fretting behavior of Ti-6Al-4V.     

Effect of different Ti-6Al-4V surface treatments on osteoblasts behaviour

The purpose of the present work was to examine the effect of different Ti-6Al-4V surface treatments on osteoblasts behaviour. Previous work in this laboratory has demonstrated that an ageing treatment reduces metal ion release from this alloy compared to standard passivation procedures. In this study. human osteosarcoma MG-63 were used in short-term in vitro tests to assay for cell viability and cell proliferation at 12, 24 and 72 h while SaOS-2 were used in long-term in vitro tests to assay for osteonectin, osteopontin, osteocalcin gene expression, total protein amount (TP). alkaline phosphatase activity (ALP) and fibronectin production (FN) for 1-4 weeks. Epifluorescence microscopy was used to observe SaOS-2 cell morphology. After 24h, there was no difference in MG-63 cell viability proliferation or in SaOS-2 cell morphology between the different surface treatments. For the long-term tests, the aged Ti-6Al4V induced significantly higher cell proliferation than the control Ti-6Al-4V at 72h. At week 1, no difference in the osteonectin, osteopontin, and osteocalcin gene expression was found between samples. The peak of ALP activity appeared earlier at week 2 for the control surface compared with the passivated and aged surfaces. The early increase in ALP activity for the control sample could be a compensatory effect of decreased osteoblasts proliferation. There was no difference in the expression of FN for the different surface treatments. Our present results showed that the different surface treatments, which induced different metal ion release kinetics and surface properties, influenced the cell proliferation and ALP activity of osteoblast cells. Aluminium ions release kinetics as well as presence of vanadium ions may play a major role in influencing the osteoblasts behaviour in the present study.     

Effects of topographical surface modifications of electron beam melted Ti-6Al-4V titanium on human fetal osteoblasts

The aim of the study was to assess the suitability of different Ti-6Al-4V surfaces produced by the electron beam melting (EBM) process as matrices for attachment, proliferation, and differentiation of human fetal osteoblasts (hFOB 1.19). Human osteoblasts were cultured in vitro on smooth and rough-textured Ti-6Al-4V alloy disks. By means of cell number and vitality and SEM micrographs cell attachment and proliferation were observed. The differentiation rate was examined by using quantitative real-time PCR analysis for the gene expression of alkaline phosphatase (ALP), type I collagen (Coll-I), bone sialoprotein (BSP) and osteocalcin (OC). After 3 days of incubation there was a significant higher vitality (p < 0.02) and proliferation (p < 0.02) of hFOB cells on smooth surfaces (R(a) = 0.077 microm) and compact surfaces with adherent partly molten titanium particles on the surface (R(a) /= 56.9 microm) reduced proliferation of hFOB cells. Surface characteristics of titanium can easily be changed by EBM in order to further improve proliferation.     

Functionally graded Co-Cr-Mo coating on Ti-6Al-4V alloy structures

Functionally graded, hard and wear-resistant Co–Cr–Mo alloy was coated on Ti–6Al–4V alloy with a metallurgically sound interface using Laser Engineering Net Shaping (LENS™). The addition of the Co–Cr–Mo alloy onto the surface of Ti–6Al–4V alloy significantly increased the surface hardness without any intermetallic phases in the transition region. A 100% Co–Cr–Mo transition from Ti–6Al–4V was difficult to produce due to cracking. However, using optimized LENS™ processing parameters, crack-free coatings containing up to 86% Co–Cr–Mo were deposited on Ti–6Al–4V alloy with excellent reproducibility. Human osteoblast cells were cultured to test in vitro biocompatibility of the coatings. Based on in vitro biocompatibility, increasing the Co–Cr–Mo concentration in the coating reduced the live cell numbers after 14 days of culture on the coating compared with base Ti–6Al–4V alloy. However, coated samples always showed better bone cell proliferation than 100% Co–Cr–Mo alloy. Producing near net shape components with graded compositions using LENS™ could potentially be a viable route for manufacturing unitized structures for metal-on-metal prosthetic devices to minimize the wear-induced osteolysis and aseptic loosening that are significant problems in current implant design.     

Effect of fluoride and albumin concentration on the corrosion behavior of Ti-6Al-4V alloy

The purpose of this study was to investigate the effect of fluoride and albumin concentration on the corrosion behavior of Ti-6Al-4V alloy in acid artificial saliva (pH 5/37 degrees C) using potentiodynamic polarization and linear polarization test. Chemical analyses of the surface passive film were characterized using X-ray photoelectron spectroscopy. The results showed that either the NaF (0-0.5%) or the presence of albumin (0-0.2%) in 0.1% NaF-containing media had a significant influence on the corrosion potential (E(corr)), corrosion rate (I(corr)), passive current density (I(pass)), and polarization resistance (R(p)) (p<0.01). The I(corr) and I(pass) values increased on increasing the NaF concentration, but decreased with the presence of albumin in NaF-containing media. The R(p) value decreased on increasing the NaF concentration, but increased with the presence of albumin in NaF-containing media. When the NaF concentration was higher than 0.1%, the protectiveness of TiO(2) passive film formed on Ti-6Al-4V alloy was destroyed by fluoride ions, leading to the formation of Na(2)TiF(6). The presence of albumin, regardless of the concentration, in 0.1% NaF-containing acid media improved the corrosion resistance of Ti-6Al-4V alloy.     

Effect of thermal oxidation on corrosion and corrosion-wear behaviour of a Ti-6Al-4V alloy

In this study, comparative investigation of thermal oxidation treatment for Ti-6Al-4V was carried out to determine the optimum oxidation conditions for further evaluation of corrosion-wear performance. Characterization of modified surface layers was made by means of microscopic examinations, hardness measurements and X-ray diffraction analysis. Optimum oxidation condition was determined according to the results of accelerated corrosion tests made in 5m HCl solution The examined Ti-6Al-4V alloy exhibited excellent resistance to corrosion after oxidation at 600 degrees C for 60 h. This oxidation condition achieved 25 times higher wear resistance than the untreated alloy during reciprocating wear test conducted in a 0.9% NaCl solution.     

Electrochemical studies of the corrosion behaviour of titanium and the Ti-6Al-4V alloy using electrochemical impedance spectroscopy

OBJECTIVES: The aim of this study was to compare the electrochemical behaviour of two materials used in oral implantology. METHODS: The resistance to corrosion of Ti grade 2 and the alloy Ti-6Al-4V was studied in an artificial saliva solution. It has been observed that the passivation of titanium by an oxidised layer can be shown both by cyclic voltametry and by electrochemical impedance spectroscopy. Moreover, this latter technique, rarely used in odontology, opens up interesting perspectives, enabling a more quantitative approach to the resistance of the passive layer to be adopted. RESULTS: Also, the impedance data recorded for Ti grade 2 and the alloy Ti-6Al-4V, in the artificial saliva solution were shown that Ti grade 2 has a higher resistance to corrosion and a thicker oxide layer than the alloy Ti-6Al-4V. SIGNIFICANCE: The fact that the electrochemical properties of Ti-6Al-4V are lower than that titanium's ones indicate than a release of ions aluminum and or vanadium ions in the body can occur. This is why we recommend to the dental practitioners to preferably use titanium in implantology.     

Porous Ti-6Al-4V alloy fabricated by spark plasma sintering for biomimetic surface modification

Porous compacts with both biological and biomechanical compatibilities and high strength were developed. Spherical powders of Ti-6Al-4V alloy, which were either as received or surface modified with the use of calcium ions by hydrothermal treatment (HTT), were fabricated by a spark plasma sintering process. The porous compacts of pure Ti were used as reference materials. Porosity was approximately 30%, and compressive strengths were 113 and 125 MPa for the as-received Ti alloy powders and those modified by the HTT process, respectively. The bending strength and elastic modulus of as-received Ti alloy powders were 128-178 MPa and 16-18 GPa, respectively. Each of the compacts was immersed in simulated body fluid (SBF). The amount of adsorption/precipitation of calcium phosphate through the compacts was measured by weight change and was observed by SEM. The compacts were covered with calcium phosphate after 2 weeks of immersion in SBF. The compacts of Ti alloy had plenty of precipitated apatite crystals, and modification by HTT accumulated more precipitation. Because calcium phosphate is a mineral component of bone, apatite, which is precipitated on the surface of the compacts, could adsorb proteins and/or drugs such as antibiotics. It is expected that a large amount of proteins and/or drugs could be impregnated when the porous compacts developed are used.     

Characterization of hydroxyapatite and titanium coatings sputtered on Ti-6Al-4V substrate.

A series of thin (<10 microm), single-layered HA/Ti coatings were deposited on Ti-6Al-4V substrate using a radio frequency magnetron-assisted sputtering system. The adhesion strength, microstructure, and chemistry of the coatings were characterized. Experimental results showed that higher Ti contents in targets or coatings resulted in higher deposition rates. When Ti was added the highly crystalline structure of monolithic HA coating was largely disrupted and the coating became amorphous-like. The highly crystalline structure of the monolithic Ti coating was also disrupted by introducing small amounts of Ca, P, and O into the coating. The HA/Ti coatings had quite uniform thicknesses and appeared smooth, dense, and well bonded to the substrate. A scanning electron microscope with an energy dispersive spectroscopy system showed that monolithic HA, 95HA/5Ti, 25HA/75Ti, and 50HA/50Ti coatings had the lowest Ca/P ratios while the 75HA/25Ti coating had the highest. The adhesion strengths of all coatings were between 60 and 80 MPa.     

Electrochemical investigation of chromium oxide-coated Ti-6Al-4V and Co-Cr-Mo alloy substrates

Hard coatings for articulating surfaces of total joint replacements may improve the overall wear resistance. However, any coating approach must take account of changes in corrosion behavior. This preliminary assessment analyzes the corrosion kinetics, impedance and mechanical-electrochemical stability of 100 μm thick plasma sprayed chromium oxide (Cr?O?) coatings on bearing surfaces in comparison to the native alloy oxide films on Co-Cr-Mo and Ti-6Al-6V. Cyclic potentiodynamic polarization, electrochemical impedance spectroscopy, and mechanical abrasion under potentiostatic conditions were performed on coated and substrate surfaces in physiological saline. SEM analysis characterized the coating morphology. The results showed that the corrosion current density values of chromium oxide coatings (0.4-1.2 μA/cm2) were of the same order of magnitude as Ti-6Al-4V alloy. Mechanical abrasion did not increase corrosion rates of chromium oxide coatings but did for uncoated Co-Cr-Mo and Ti-6Al-4V. The impedance response of chromium oxide coatings was very different than Co-Cr-Mo and Ti-6Al-4V native oxides characterized by a defected coating model. More of a frequency-independent purely resistive response was seen in mid-frequency range for the coatings (CPE(coat) : 40-280 nF/cm2 (rad/s)(1-α) , α: 0.67-0.83) whereas a more capacitive character is seen for Co-Cr-Mo and Ti-6Al-4V (CPE(ox) around 20 μF/cm2 (rad/s)(1-α) , α around 0.9). Pores, interparticle gaps and incomplete fusion typical for thermal spray coatings were present in these oxides which could have influenced corrosion resistance. The coating microstructure could have allowed some fluid penetration. Overall, these coatings appear to have suitable corrosion properties for wear surfaces.     

Wear-accelerated corrosion of Ti-6Al-4V and nitrogen-ion-implanted Ti-6Al-4V: mechanisms and influence of fixed-stress magnitude

Wear-accelerated corrosion rates at constant anodic potentials were evaluated for unimplanted and nitrogen-ion-implanted surgical Ti-6Al-4V while wearing against ultrahigh-molecular-weight polyethylene at stress levels up to 6.90 MPa (1000 psi). The ion implantation processing was found to reduce the wear corrosion rates in both saline and serum solutions at all applied stress levels. During wear testing, all of the ion-implanted surfaces remained visually unchanged from the polished condition. However, many of the unimplanted surfaces developed damage zones characterized by wear tracks and black wear debris. A surface-damage mechanism is proposed and discussed which involves disruption of the Ti-6Al-4V protective oxide film, subsequent entrapment of oxide particles in the polyethylene, then self-perpetuating damage due to the abrasive action of the embedded particles.     

In vitro biocorrosion of Ti-6Al-4V implant alloy by a mouse macrophage cell line

Corrosion of implant alloys releasing metal ions has the potential to cause adverse tissue reactions and implant failure. We hypothesized that macrophage cells and their released reactive chemical species (RCS) affect the alloy's corrosion properties. A custom cell culture corrosion box was used to evaluate how cell culture medium, macrophage cells and RCS altered the Ti-6Al-4V corrosion behaviors in 72 h and how corrosion products affected the cells. There was no difference in the charge transfer in the presence (75.2 +/- 17.7 mC) and absence (62.3 +/- 18.8 mC) of cells. The alloy had the lowest charge transfer (28.2 +/- 4.1 mC) and metal ion release (Ti < 10 ppb, V < 2 ppb) with activated cells (releasing RCS) compared with the other two conditions. This was attributed to an enhancement of the surface oxides by RCS. Metal ion release was very low (Ti < 20 ppb, V < 10 ppb) with nonactivated cells and did not change cell morphology, viability, and NO and ATP release compared with controls. However, IL-1beta released from the activated cells and the proliferation of nonactivated cells were greater on the alloy than the controls. In summary, macrophage cells and RCS reduced the corrosion of Ti-6Al-4V alloys as hypothesized. These data are important in understanding host tissue-material interactions.     

Varying Ti-6Al-4V surface roughness induces different early morphologic and molecular responses in MG63 osteoblast-like cells

Osteoblast response to Ti implants depends not only on the chemistry of the implant but also on the physical properties of the implant surface, such as microtopography and roughness. This study was undertaken to examine early changes in cell morphology and gene expression during the early phase of osteoblast interaction with titanium alloy (Ti-6Al-4V) surfaces of two different roughnesses. MG63 osteoblast-like cells were cultured for 2, 6, 24, and 72 h on smooth (Ra=0.18+/-0.03 microm) and rough (Ra=2.95+/-0.23 microm) Ti-6Al-4V surfaces. Changes in cell proliferation were assessed by measuring cell number after 72 h in culture. Morphological characteristics were observed by scanning electron microscopy after 2, 6, and 24 h of culture. Changes in gene expression for extracellular signal-regulated kinase 2 (Erk2), type I collagen (alpha2[I] collagen), phospholipase C-gamma2 (Plc-gamma2), and beta-actin were measured by RT-PCR after 6 and 24 h in culture. Cell number was significantly higher on the smooth surface. In scanning electron micrographs, cells on smooth Ti-6Al-4V were spherical and raised up from the surface after 2 h in culture. In contrast, cells on the rough surface adopted an irregular, elongated shape that spanned across pits in the surface. At 24 h, cells on the smooth surface had flattened, become elongate, and covered the surface. In contrast, cells on the rough surface appeared more differentiated in shape and the margins of the cells were irregular, with many processes extending out, following the contour of the surface. Of the genes examined, only Erk2 and beta-actin showed a change in expression with surface roughness. Both genes were upregulated (p<0.05) on the rough surface at 6 h. These results indicate that Ti-6Al-4V surface roughness affects osteoblast proliferation, morphology, and gene expression, and that these effects can be measured after periods as short as 2-6 h.     

The corrosion behaviour of Ti-6Al-4V, Ti-6Al-7Nb and Ti-13Nb-13Zr in protein solutions

Ti alloys are used in orthopaedic applications owing to their appropriate mechanical properties and their excellent corrosion resistance. The release of titanium and the other alloying elements into the surrounding tissue has been reported due either to passive corrosion or accelerating processes such as wear. Since the passive layer can be broken down in certain circumstances by wear it is important to study the ability of these alloys to repassivate in biological environments, in particular in the presence of proteins, and evaluate how the repassivated surface may vary from the original surface. In this study we investigated the ability of Ti-6Al-4V, Ti-6Al-7Nb and Ti-13Nb-13Zr to repassivate in phosphate buffered saline (PBS), bovine albumin solutions in PBS and 10% foetal calf serum in PBS at different pH values and at different albumin concentrations. It was found that an increase in pH had a greater effect on the corrosion behaviour of Ti-6Al-4V and Ti-6Al-7Nb than on Ti-13Nb-13Zr in PBS and that the addition of protein to the PBS reduced the influence of pH on the corrosion behaviour of all the alloys. The effect of the corrosion and repassivation was investigated by measuring changes in the surface hardness of the alloys and it was found that corrosion reduced the hardness of the surface oxides of all the alloys. In PBS the reduction was smallest for Ti-6Al-4V and largest for Ti-13Nb- 3Zr and that corrosion in protein solutions further reduced the hardness of the surface oxides. This effect was greater for Ti-6Al-4V and Ti-6Al-7Nb than for Ti-13Nb-13Zr. In conclusion, proteins in the environment appear to interact with the repassivation process at the surface of these alloys and influence the resulting surface properties.     

Acoustic emission during fatigue of porous-coated Ti-6Al-4V implant alloy.

Acoustic emission (AE) events and event intensities (e.g., event amplitude, counts, duration, and energy counts) were recorded and analyzed during fatigue loading of uncoated and porous-coated Ti-6Al-4V. AE source location, spatial filtering, event, and event intensity distributions were used to detect, monitor, analyze, and predict failures. AE provides the ability to spatially and temporally locate multiple fatigue cracks, in real time. Fatigue of porous-coated Ti-6Al-4V is governed by a sequential, multimode fracture process of: transverse fracture in the porous coating; sphere/sphere and sphere/substrate debonding; substrate fatigue crack initiation; slow and rapid substrate fatigue crack propagation. Because of the porosity of the coating, the different stages of fracture within the coating occur in a discontinuous fashion. Therefore, the AE events generated are intermittent and the onset of each mode of fracture in the porous coating can be detected by increases in AE event rate. Changes in AE event rate also correspond to changes in crack extension rate, and may therefore be used to predict failure. AE offers two distinct advantages over conventional optical and microscopic methods of analyzing fatigue cracks--it is more sensitive and it can determine the time history of damage progression. The magnitude of the AE event intensities increased with increasing stress. Failure mechanisms are best differentiated by analyzing AE event amplitudes. Intergranular fracture and microvoid coalescence generated the highest AE event amplitudes (100 dB), whereas, plastic flow and friction generated the lowest AE event amplitudes (55-65 dB). Fractures in the porous coating were characterized by AE event amplitudes of less than 80 dB.