Bioactivity of sol-gel-derived TiO2 coating on polyetheretherketone: In vitro and in vivo studies.

A polyetheretherketone (PEEK) surface was modified using a sol-gel-derived TiO2 coating in order to confer bone-bonding ability. To enhance the bonding strength of the coating layer, pretreatment with either O2 plasma or sandblasting was performed prior to sol-gel coating. Additionally, post-treatment with acid was carried out to confer apatite (calcium phosphate)-forming ability to the surface. Biomechanical and histological analyses performed using an in vivo rabbit tibia model showed that PEEK surfaces modified with sol-gel-derived TiO2 and acid post-treatment had better bone-bonding properties than uncoated PEEK surfaces. These modified surfaces also performed well in terms of their in vitro cell responses due to their modified surface chemistries and topographies. Although O2 plasma or sandblasting treatment were, for the most part, equivocal in terms of performance, we conclude that sol-gel-derived TiO2 coating followed by acid post-treatment significantly improves the bone bonding ability of PEEK surfaces, thus rendering them optimal for their use in surgical implants. STATEMENT OF SIGNIFICANCE: The role of polyetheretherketone (PEEK) as an alternative biomaterial to conventional metallic implant materials has become increasingly important. However, its low bone bonding ability is yet to be resolved. This in vivo and in vitro investigation on the functionalization of PEEK surfaces highlights the utility of this material in clinical interventions that require implants, and may extend range of applications of PEEK.

Apatite-forming PEEK with TiO2 surface layer coating.

Polyetheretherketone (PEEK) is widely used in orthopedic implants, such as spinal fusion devices, because of its moderate elastic modulus, as well as relatively high mechanical strength. However, it does not bond to living bone, and hence it needs autograft to be fixed to the bone. In this study, we attempted to add bone-bonding properties to PEEK by coating with TiO2 synthesized by the sol-gel process. When a TiO2 sol solution consisting of titanium isopropoxide, water, ethanol, and nitric acid was deposited on a PEEK substrate without any pretreatment, the formed TiO2 gel layer was easily peeled off after subsequent treatments. However, when the same solution was deposited on PEEK that was preliminarily subjected to UV or O2 plasma treatment, the deposited TiO2 gel layer strongly adhered to the substrate even after subsequent treatments. The strong adhesion was attributed to the interaction among the C-O, C=O, and O-C=O groups on the PEEK owing to the UV or O2 plasma treatment and the Ti-O bond of the TiO2 gel. Apatite did not form on the as-formed TiO2 gel layer in a simulated body fluid (SBF) even within 3 days; however, apatite formed after soaking in 0.1 M HCl solution at 80 °C for 24 h. This apatite formation was attributed to positive surface charge of the TiO2 gel layer induced by the acid treatment. The PEEK with the TiO2 gel layer coating formed by the proposed process is expected to bond to living bone, because a positively charged titanium oxide which facilitates the formation of apatite in SBF within a short period is known to bond to living bone.

Antibacterial and bioactive calcium titanate layers formed on Ti metal and its alloys.

An antibacterial and bioactive titanium (Ti)-based material was developed for use as a bone substitute under load-bearing conditions. As previously reported, Ti metal was successively subjected to NaOH, CaCl2, heat, and water treatments to form a calcium-deficient calcium titanate layer on its surface. When placed in a simulated body fluid (SBF), this bioactive Ti formed an apatite layer on its surface and tightly bonded to bones in the body. To address concerns regarding deep infection during orthopedic surgery, Ag(+) ions were incorporated on the surface of this bioactive Ti metal to impart antibacterial properties. Ti metal was first soaked in a 5 M NaOH solution to form a 1 µm-thick sodium hydrogen titanate layer on the surface and then in a 100 mM CaCl2 solution to form a calcium hydrogen titanate layer via replacement of the Na(+) ions with Ca(2+) ions. The Ti material was subsequently heated at 600 °C for 1 h to transform the calcium hydrogen titanate into calcium titanate. This heat-treated titanium metal was then soaked in 0.01-10 mM AgNO3 solutions at 80 °C for 24 h. As a result, 0.1-0.82 at.% Ag(+) ions and a small amount of H3O(+) ions were incorporated into the surface calcium titanate layers. The resultant products formed apatite on their surface in an SBF, released 0.35-3.24 ppm Ag(+) ion into the fetal bovine serum within 24 h, and exhibited a strong antibacterial effect against Staphylococcus aureus. These results suggest that the present Ti metals should exhibit strong antibacterial properties in the living body in addition to tightly bonding to the surrounding bone through the apatite layer that forms on their surfaces in the body.

Bone bonding ability of a chemically and thermally treated low elastic modulus Ti alloy: gum metal.

The gum metal with composition Ti-36Nb-2Ta-3Zr-0.3O, is free from cytotoxic elements and exhibits a low elastic modulus as well as high mechanical strength. We have previously demonstrated that this gum metal, once subjected to a series of surface treatments--immersion in 1 M NaOH (alkali treatment) and then 100 mM CaCl2, before heating at 700 °C (sample: ACaH-GM), with an optional final hot water immersion (sample: ACaHW-GM)--has apatite-forming ability in simulated body fluid. To confirm the in vivo bioactivity of these treated alloys, failure loads between implants and bone at 4, 8, 16, and 26 weeks after implantation in rabbits' tibiae were measured for untreated gum metal (UT-GM), ACaH-GM and ACaHW-GM, as well as pure titanium plates after alkali and heat treatment (AH-Ti). The ACaH-GM and UT-GM plates showed almost no bonding, whereas ACaHW-GM and AH-Ti plates showed successful bonding by 4 weeks, and their failure loads subsequently increased with time. The histological findings showed a large amount of new bone in contact with the surface of ACaHW-GM and AH-Ti plates, suggesting that the ACaHW treatment could impart bone-bonding bioactivity to a gum metal in vivo. Thus, with this improved bioactive treatment, these advantageous gum metals become useful candidates for orthopedic and dental devices.

Effect of Ca contamination on apatite formation in a Ti metal subjected to NaOH and heat treatments.

It has long been known that titanium (Ti) metal bonds to living bone through an apatite layer formed on its surface in the living body after it had previously been subjected to NaOH and heat treatments and as a result had formed sodium titanate on its surface. These treatments were applied to a porous Ti metal layer on a total hip joint and the resultant joint has been in clinical use since 2007. It has been also demonstrated that the apatite formation on the treated Ti metal in the living body also occurred in an acelullar simulated body fluid (SBF) with ion concentrations nearly equal to those of the human blood plasma, and hence bone-bonding ability of the treated Ti metal can be evaluated using SBF in vitro. However, it was recently found that certain Ti metals subjected to the same NaOH and heat treatments display apatite formation in SBF which is decreased with the increasing volume of the NaOH solution used in some cases. This indicates that bone-bonding ability of the treated Ti metal varies with the volume of the NaOH solution used. In the present study, this phenomenon was systematically investigated using commercial NaOH reagents and is considered in terms of the structure and composition of the surface layers of the treated Ti metals. It was found that a larger amount of the calcium contamination in the NaOH reagent is concentrated on the surface of the Ti metal during the NaOH treatment with an increasing volume of the NaOH solution, and that this inhibited apatite formation on the Ti metal in SBF by suppressing Na ion release from the sodium titanate into the surrounding fluid. Even a Ca contamination level of 0.0005 % of the NaOH reagent was sufficient to inhibit apatite formation. On the other hand, another NaOH reagent with a nominal purity of just 97 % did not exhibit any such inhibition, since it contained almost no Ca contamination. This indicates that NaOH reagent must be carefully selected for obtaining reliable bone-bonding implants of Ti metal by the NaOH and heat treatments.

Formation of a bioactive calcium titanate layer on gum metal by chemical treatment.

The so-called gum metal with the composition Ti-36Nb-2Ta-3Zr-0.3O is free from cytotoxic elements and exhibits a low elastic modulus as well as high mechanical strength. In the present study, it was shown that this alloy exhibited a high capacity for apatite formation in a simulated body fluid when subjected to 1 M NaOH treatment, 100 mM CaCl(2) treatment, heat treatment at 700°C, and then hot water treatment. The high apatite formation was attributed to the CaTi(2)O(5) which was precipitated on its surface, and found to be maintained even in a humid environment over a long period. The treated surface exhibited high scratch resistance, which is likely to be useful in clinical applications. The surface treatment had little effect on the unique mechanical properties described above. These results show that gum metal subjected to the present surface treatments exhibits a high potential for bone-bonding, which will be useful in orthopedic and dental implants.

Effect of titania-based surface modification of polyethylene terephthalate on bone-implant bonding and peri-implant tissue reaction.

Organic polymers can be uniformly surface-modified with bioactive TiO(2) by using a sol-gel method. Titania-based surface-modified polyethylene terephthalate (TiPET) plates and fabric have shown apatite-forming ability in simulated body fluid. Here, we first investigated the bone-bonding ability and mechanical bonding strength between the surface-modified layer and the base material (PET) of TiPET plates in vivo. For clinical applicability, we also examined the bone-bonding ability of TiPET fabric and the effect of titania-based surface modification on peri-implant tissue reactions (e.g. connective tissue capsule formation) in bone in vivo. Solid PET plates and PET fabric were prepared. Test plates and fabric were surface-modified with titania solution by using a sol-gel method. Histological examinations of the plates implanted into rabbit tibiae revealed direct contact between the TiPET plate and the bone. After the detaching test, a considerable amount of bone residue was observed on the surface of the TiPET plate. This result suggests that the mechanical bond strength between surface-modified layer and the base material is stronger than that between newly generated bone and tibia, and indirectly ensures the mechanical stability of the surface-modified layer. Pulling tests and histological examinations of the TiPET fabric revealed its excellent bone-bonding ability and micro-computed tomographic images showed excellent osteoconductive ability of TiPET fabric. The connective tissue capsule was much thinner, with less inflammatory tissue around the TiPET implants than around the control samples. These results indicate that TiPET fabric possesses a mechanically stable surface-modified layer, excellent bone-bonding ability, osteoconductive ability, and biocompatibility in bone.

Preparation of bioactive Ti metal surface enriched with calcium ions by chemical treatment.

A calcium solution treatment was applied to a NaOH-treated titanium metal to give it bioactivity, scratch resistance and moisture resistance. The titanium metal was soaked in a 5 M NaOH solution and then a 100 mM CaCl(2) solution to incorporate Ca(2+) ions into the titanium metal surface by ion exchange. This treated titanium metal was subsequently heated at 600 degrees C and soaked in hot water at 80 degrees C. The NaOH treatment incorporated approximately 5 at.% Na(+) ions into the Ti metal surface. These Na(+) ions were completely replaced by Ca(2+) ions by the CaCl(2) treatment. The number of Ca(2+) ions remained even after subsequent heat and water treatments. Although the NaOH-CaCl(2)-treated titanium metal showed slightly higher apatite-forming ability in a simulated body fluid than the NaOH-treated titanium metal, it lost its apatite-forming ability during the heat treatment. However, subsequent water or autoclave treatment restored the apatite-forming ability of the NaOH-CaCl(2)-heat-treated titanium metal. Although the apatite-forming ability of the NaOH-heat-treated titanium metal decreased dramatically when it was kept at high humidity, that of NaOH-CaCl(2)-heat-water-treated titanium metal was maintained even in the humid environment. The heat treatment increased the critical scratch resistance of the surface layer of the NaOH-CaCl(2)-treated titanium metal remarkably, and it did not deteriorate on subsequent water treatment.

Specific response of osteoblast-like cells on hydroxyapatite layer containing serum protein.

Accelerations of bone-like apatite deposition and cell growth on an electrically polarized ceramic hydroxyapatite have been reported. A relationship between these phenomena was investigated in a previous report, and then it was suggested that osteoblast-like cell's (MC3T3-E1) growth had relevance to the mineral growth. The effect of the formed apatite layer especially appeared to be on the cell adhesion. The acceleration of cell proliferation on the polarized HAp has been shown using fibroblastic cell (L929) and nerve cell (SK-N-SH) lines, therefore the effect of the layer on L929 and SK-N-SH was investigated to support the mechanism of acceleration of cell proliferation by polarization of HAp. In this study, the effect of the bone-like apatite layer was not confirmed on L929 cell's growth. On the other hand, the acceleration of nerve cell's proliferation was confirmed on the formed apatite layer. However, the remarkable improvement of the cell adhesion of SK-N-SH was not confirmed on the apatite layer. Consequently, it was considered that the bone-like apatite containing serum protein obtained by the coprecipitation of bone-like apatite and serum protein has a pronounced role only in the activity of osteoblast-like cells.

Effect of bone-like layer growth from culture medium on adherence of osteoblast-like cells.

The electrical polarization of ceramic HAp had an effect on the acceleration of bone restoration. Cell behavior in the bone-like growth layer was investigated. The deposits on the ceramic HAp was grown and formed layers by soaking in alpha-minimum essential medium supplemented with 10% fetal bovine serum (alpha-MEM supplemented with 10% FBS). The shapes of the adhering cells on the grown layer gradually changed from spindle to flat with growth of the layer. On the totally grown layer that was grown on the ceramic HAp by soaking in alpha-MEM supplemented with 10% FBS for 7 days, all the adhering cells were flat and the surface was filled with the grown cells. From these results, it was revealed that the grown layer on the ceramic HAp is one of the activation factors of cell growth. Consequently, cell growth was reinforced by acceleration of the layer growth on the negatively charged surface of the polarized ceramic HAp.