In vitro bioactivity evaluation of titanium and niobium metals with different surface morphologies

Current orthopaedic biomaterials research mainly focuses on designing implants that could induce controlled, guided and rapid healing. In the present study, the surface morphologies of titanium (Ti) and niobium (Nb) metals were tailored to form nanoporous, nanoplate and nanofibre-like structures through adjustment of the temperature in the alkali-heat treatment. The in vitro bioactivity of these structures was then evaluated by soaking the treated samples in simulated body fluid (SBF). It was found that the morphology of the modified surface significantly influenced the apatite-inducing ability. The Ti surface with a nanofibre-like structure showed better apatite-inducing ability than the nanoporous or nanoplate surface structures. A thick dense apatite layer formed on the Ti surface with nanofibre-like structure after 1 week of soaking in SBF. It is expected that the nanofibre-like surface could achieve good apatite formation in vivo and subsequently enhance osteoblast cell adhesion and bone formation.

Bone formation following implantation of titanium sponge rods into humeral osteotomies in dogs : a histological and histometrical study

Background: Titanium (Ti) is widely proven to enhance bone contact and growth on its surface. It is expected that bone defects could benefit from Ti to promote healing and to increase strength of the implanted area.

Purpose: The present study aimed at comparing the potential of porous Ti sponge rods with synthetic hydroxyapatite (HA) for the healing of bone defects in a canine model.

Material and Methods: Six mongrel dogs were submitted to three trephined osteotomies of 6.0 × 4.0 mm in one humerus and after 2 months another three osteotomies were performed in the contralateral humerus. A total of 36 defects were randomly filled either with Ti foam, particulate HA, or coagulum (control). The six animals were killed 4 months after the first surgery for histological and histometrical analysis.

Results: The Ti-foam surface was frequently found in intimate contact with new bone especially at the defect walls. Control sites showed higher amounts of newly formed bone at 2 months – Ti (p = 0.000) and HA (p = 0.009) – and 4 months when compared with Ti (p = 0.001). Differently from HA, the Ti foam was densely distributed across the defect area which rendered less space for bone growth in the latter's sites. The use of Ti foams or HA resulted in similar amounts of bone formation in both time intervals. Nevertheless, the presence of a Ti-foam rod preserved defect's marginal bone height as compared with control groups. Also, the Ti-foam group showed a more mature bone pattern at 4 months than HA sites.

Conclusion: The Ti foam exhibited good biocompatibility, and its application resulted in improved maintenance of bone height compared with control sites. The Ti foam in a rod design exhibited bone ingrowth properties suitable for further exploration in other experimental situations.

Mechanical properties of titanium foam for biomedical applications

Understanding the mechanical behaviour of pure titanium (Ti) foam is crucial for the design and development of Ti foam-based load-bearing implants. In this work, pure titanium foam is fabricated by a powder metallurgical process using the space-holder technique with a spacer size of 500 to 800 µm. Experimental data from static compression testing on the Ti foam are presented. The application of theoretical formulae to predict Young's modulus and yield strength of titanium foams is also discussed. A foam with 63% porosity, 87 ± 5 MPa yield strength, and 6.5 ± 1.3 GPa Young's modulus is found to be appropriate for a number of dental and orthopaedic applications.

Influence of calcium ion deposition on apatite-inducing ability of porous titanium for biomedical applications

In the present study, the influence of calcium ion deposition on the apatite-inducing ability of porous titanium(Ti) was investigated in a modified simulated body fluid (m-SBF). Calcium hydroxide (Ca(OH)₂) solutions with five degrees of saturation were used to hydrothermally deposit Ca ions on porous Ti with a porosity of 80%. Apatite-inducing ability of the Ca-ion-deposited porous Ti was evaluated by soaking them in m-SBF for up to 14 days. Scanning electron microscopy (SEM) and X-ray diffractometry (XRD) confirmed that a thin layer of calcium titanate (CaTiO₃)/calcium oxide (CaO) mixture with a nanostructured porous network was produced on porous Ti substrates after hydrothermal treatment at 200 °C for 8 h. X-ray photoelectron spectroscopy results demonstrated that the content of the Ca ions deposited on Ti and the thickness of the CaTiO₃/CaO layer increased with increasing saturation degree of the Ca(OH)₂ solution. The thickest (over 10 nm) CaTiO₃/CaO layer with the highest Ca content was achieved on the Ti treated in an oversaturated Ca(OH)₂ solution (0.2 M). SEM, XRD, transmission electron microscopy and Fourier transformed infrared spectroscopy analysis indicated that the porous Ti samples deposited with the highest content of Ca ions exhibited the best apatite-inducing ability, producing a dense and complete carbonated apatite coating after a 14 day soaking in m-SBF. The present study illustrated the validity of using Ca ion deposition as a pre-treatment to endow desirable apatite-inducing ability of porous Ti for bone tissue engineering applications.

Microstructures and bond strengths of the calcium phosphate coatings formed on titanium from different simulated body fluids

Calcium phosphate (Ca-P) coatings were deposited on Ti substrates by a biomimetic method from m-SBF and 10× SBF, respectively. Comparative study of microstructures and bond strengths of the Ca-P coatings deposited from those different SBFs was carried out. Effect of the surface roughness of the substrates on the bond strength of the Ca-P coatings was also studied. Scanning electron microscopy (SEM), X-ray diffractometry (XRD), Fourier transformed infrared spectroscopy (FTIR), inductive coupled plasma spectrometry (ICP) and thermogravimetry (TG) were used to characterize the Ca-P coatings. The bond strengths between the coatings and Ti substrates were measured using an adhesive strength test. Results indicated that the ionic concentrations of the SBFs and the surface roughness of the substrate had a significant influence on the formation, morphology and bond strength of the Ca-P precipitates. The induction period of time to deposit a complete Ca-P layer from the m-SBF is much longer, but the Ca-P coating is denser and has higher bond strength than that formed from the 10× SBF. The Ti with a surface roughness of R_a 0.64 µm and R_z 2.81 µm favoures the formation of a compact Ca-P coating from the m-SBF with the highest bond strength of approximately 15.5 MPa.

The importance of particle size in porous titanium and nonporous counterparts for surface energy and its impact on apatite formation

The importance of particle size in titanium (Ti) fabricated by powder metallurgy for the surface energy and its impact on the apatite formation was investigated. Four sorts of Ti powders of different mean particle size were realized through 20 min, 2 h, 5 h and 8 h of ball milling, respectively. Each sort of Ti powder was used to fabricate porous Ti and its nonporous counterparts sharing similar surface morphology, grain size and chemical composition, and then alkali-heat treatment was conducted on them. Surface energy was measured on the surfaces of the nonporous Ti counterparts due to the difficulty in measuring the porous surfaces directly. The surface energy increase on the alkali-heat-treated porous and nonporous Ti was observed due to the decrease in the particle size of the Ti powders and the presence of Ti–OH groups brought by the alkali-heat treatment. The apatite-inducing ability of the alkali-heat-treated porous and nonporous Ti with different surface energy values was evaluated in modified simulated body fluid and results indicated that there was a strong correlation between the apatite-inducing ability and the surface energy. The alkali-heat-treated porous and nonporous Ti discs prepared from the powders with an average particle size of 5.89 ± 0.76 μm possessed the highest surface energy and the best apatite-inducing ability when compared to the samples produced from the powders with the average particle size varying from 19.79 ± 0.31 to 10.25 ± 0.39 μm.

In vitro cytotoxicity of binary TI alloys for bone implants

Interest in using titanium (Ti) alloys as load-bearing implant materials has increased due to their high strength to weight ratio, lower elastic modulus, and superior biocompatibility and enhanced corrosion resistance compared to conventional metals such as stainless steel and Co-Cr alloys. In the present study, the in vitro cytotoxicity of five binary titanium alloys, Ti15Ta, Ti15Nb, Ti15Zr, Ti15Sn and Ti15Mo, was assessed using human osteosarcoma cell line, SaOS-2 cells. The Cell proliferation and viability were determined, and cell adhesion and morphology on the surfaces of the binary Ti alloys after cell culture were observed by SEM. Results indicated that the Ti binary alloys of Ti15Ta, Ti15Nb and Ti15Zr exhibited the same level of excellent biocompatibility; Ti15Sn alloy exhibited a moderate biocompatibility while Ti15Mo alloy exhibited a moderate cytotoxicity. The SaOS-2 osteoblast-like cells had flattened and spread across the surfaces of the Ti15Ta, Ti15Nb, Ti15Zr and Ti15Sn groups; however, the cell shapes on the Ti15Mo alloy was shrinking and unhealthy. These results indicated that the Mo contents should be limited to a certain level in the design and development of new Ti alloys for implant material applications.

Bioactivating the surfaces of titanium by sol-gel process

In the present study, titanium (Ti) samples were surface-modified by titania (TiO₂), silica (SiO₂) and hydroxyapatite (HA) coatings using a sol-gel process. The bioactivity of the film-coated Ti samples was investigated by cell attachment and morphology study using human osteoblast-like SaOS-2 cells. Results of the cell attachment indicated that the densities of cell attachment on the surfaces of Ti samples were significantly increased by film coatings; the density of cell attachment on HA film-coated surface was higher than those on TiO₂ and SiO₂ film-coated surfaces. Cell morphology study showed that the cells attached, spread and grew well on the three kinds of film-coated surfaces. It can be concluded that the three kinds of film coatings can bioactivate the surfaces of Ti samples effectively. Overall, Ti sample with HA film-coated surface exhibited the best bioactivity.

Biomimetic coating on pure titanium submitted to different surface treatments

Titanium (Ti) plates were firstly treated to form various types of oxide layers on the surface and then immersed into simulated body fluid (SBF) to evaluate the apatite forming ability. The surface morphology and roughness of the different oxide layers were measured by atomic force microscopy (AFM), and the surface energies were determined based on the Owens-Wendt (OW) methods. It was found that Ti samples after Alkali-Heat treatment (AH) achieved the best apatite formation after soaking in SBF for 3 weeks, compared to those without treatment, thermal or H₂O₂ oxidation. Furthermore, contact angle measurement revealed that the oxide layer on the alkali-heat treated Ti samples possessed the highest surface energy. The results indicate that the apatite inducing ability of a titanium oxide layer is linked to its surface energy. Apatite nucleation is easier on a surface with a higher surface energy.

Apatite formation on nano-structured titanium and niobium surface

Current orthopaedic biomaterials research mainly focuses on developing implants that could induce controlled, guided and rapid healing. In the present study, the surface morphologies of titanium (Ti) and niobium (Nb) metals were tailored to form nanoporous, nanoplate and nanofibrelike structures through adjustment of the temperature in the alkali treatment. The in vitro bioactivity of these structures was then evaluated by soaking in simulated body fluid (SBF). It was found that the morphology of the modified surface significantly influenced the apatite inducing ability. The Ti surface with a nanofiber-like structure showed better apatite inducing ability, than the nanoporous or nanoplate surface structures. A thick dense apatite layer formed on the Ti surface with nanofiberlike structure after 1 week soaking in SBF. It is expected that the anofibre-like surface could achieve good apatite formation in vivo and subsequently enhance osteoblast cell adhesion and bone formation in vivo.

Biomimetic porous titanium scaffolds for orthopaedic and dental applications

The development of artificial organs and implants for replacement of injured and diseased hard tissues such as bones, teeth and joints is highly desired in orthopedic surgery. Orthopedic prostheses have shown an enormous success in restoring the function and offering high quality of life to millions of individuals each year. Therefore, it is pertinent for an engineer to set out new approaches to restore the normal function of impaired hard tissues.

Over the last few decades, a large number of metals and applied materials have been developed with significant improvement in various properties in a wide range of medical applications. However, the traditional metallic bone implants are dense and often suffer from the problems of adverse reaction, biomechanical mismatch and lack of adequate space for new bone tissue to grow into the implant. Scientific advancements have been made to fabricate porous scaffolds that mimic the architecture and mechanical properties of natural bone. The porous structure provides necessary framework for the bone cells to grow into the pores and integrate with host tissue, known as osteointegration. The appropriate mechanical properties, in particular, the low elastic modulus mimicking that of bone may minimize or eliminate the stress-shielding problem. Another important approach is to develop biocompatible and corrosion resistant metallic materials to diminish or avoid adverse body reaction. Although numerous types of materials can be involved in this fast developing field, some of them are more widely used in medical applications. Amongst them, titanium and some of its alloys provide many advantages such as excellent biocompatibility, high strength-to-weight ratio, lower elastic modulus, and superior corrosion resistance, required for dental and orthopedic implants. Alloying elements, i.e. Zr, Nb, Ta, Sn, Mo and Si, would lead to superior improvement in properties of titanium for biomedical applications.

New processes have recently been developed to synthesize biomimetic porous titanium scaffolds for bone replacement through powder metallurgy. In particular, the space holder sintering method is capable of adjusting the pore shape, the porosity, and the pore size distribution, notably within the range of 200 to 500 m as required for osteoconductive applications. The present chapter provides a review on the characteristics of porous metal scaffolds used as bone replacement as well as fabrication processes of porous titanium (Ti) scaffolds through a space holder sintering method. Finally, surface modification of the resultant porous Ti scaffolds through a biomimetic chemical technique is reviewed, in order to ensure that the surfaces of the scaffolds fulfill the requirements for biomedical applications.

Nanocrystalline titanium to mesoporous anatase with high bioactivity

In present study, the formation of bioactive anatase on bulk titanium (Ti) by hybrid surface mechanical attrition treatment (SMAT) is reported. A commercial pure Ti plate first underwent SMAT in a vacuum for 1 h to produce a nanocrystalline layer with a thickness of about 30 µm, and then the nanocrystalline Ti (30 nm) was transformed into mesoporous anatase with a grain size 10 nm by chemical oxidation and calcination. The mesoporous anatase showed excellent bioactivity while being soaked in simulated body fluid, which could be attributed to the unique nanostructure on the SMAT Ti surface.

Interfacial chemistry and adhesion between titanium dioxide nanotube layers and titanium substrates

Interfacial chemistry and adhesion between titanium dioxide (TiO₂) nanotube layers and titanium (Ti) substrates were studied in this Article. TiO₂ nanotube layers were produced on pure Ti by anodization and annealed in air for different time durations. The adhesion of the TiO₂ nanotube layers was then investigated by Rockwell C indentation test. Results show that adhesion of TiO₂ nanotube layers improved with the extension of annealing time. This improvement in adhesion of TiO₂ nanotube layers was analyzed from the viewpoint of interfacial chemistry using energy dispersive X-ray spectrometry (EDS) and X-ray photoelectron spectroscopy (XPS). It suggests that more Ti-O bonds formed in the interface after annealing, and this led to the improved adhesion of the TiO₂ nanoube layers.

Tribological behaviour of pure Ti with a nanocrystalline surface layer under different loads

Surface mechanical attrition treatment (SMAT), a novel surface severe plastic deformation method, was carried out for titanium (Ti) to create a gradient-structured Ti (SMAT Ti). The tribological behaviour was studied under different loads and dry sliding conditions. The results showed that the deformation layer of SMAT Ti was about 160 μm. The friction and wear results showed that the wear resistance of SMAT Ti was enhanced compared to the coarse-grained (CG) counterpart. SMAT Ti showed abrasive wear under 1 and 5 N, and exhibited abrasive and adhesive wear under 2 N. While CG Ti showed abrasive and adhesive wear under 1-2 N, and exhibited abrasive wear under 5 N for the work hardening effects.

In vitro behavior of human osteoblast-like cells (SaOS2) cultured on surface modified titanium and titanium–zirconium alloy

In this study, titanium (Ti) and titanium-zirconium (TiZr) alloy samples fabricated through powder metallurgy were surface modified by alkali-heat treatment and calcium (Ca)-ion-deposition. The alteration of the surface morphology and the chemistry of the Ti and TiZr after surface modification were examined. The bioactivity of the Ti and TiZr alloys after the surface modification was demonstrated. Subsequently, the cytocompatibility of the surface modified Ti and TiZr was evaluated via in vitro cell culture using human osteoblast-like cells (SaOS2). The cellular attachment, adhesion and proliferation after cell culture for 14 days were characterized by scanning electron microscopy (SEM) and MTT assay. The relationship between surface morphology and chemical composition of the surface modified Ti and TiZr and cellular responses was investigated. Results indicated that the surface-modified Ti and TiZr alloys exhibited excellent in vitro cytocompatibility together with satisfactory bioactivity. Since osteoblast adhesion and proliferation are essential prerequisites for a successful implant in vivo, these results provide evidence that Ti and TiZr alloys after appropriate surface modification are promising biomaterials for hard tissue replacement.