Enhancing the bioactivity of zirconia and zirconia composites by surface modification

Among bioceramics, zirconia (ZrO(2)) and alumina (Al(2)O(3)) possess exceptional mechanical properties suitable for load-bearing and wear-resistant applications but the poor bioactivity of these materials is the major concern when bonding and integration to the living bone are desired. This article investigates two different approaches and their underlying mechanisms to improve the bioactivity of zirconia (3Y-TZP) and a zirconia composite with alumina (10Ce-TZP/Al(2)O(3)). Chemical treatment approach applied on 3Y-TZP where the substrates were soaked in 5M H(3)PO(4) to create chemically functional groups on the surface for inducing apatite nucleation. X-ray photoelectron spectroscopy (XPS) was used to detect chemical changes and X-ray diffraction (XRD) to monitor phase changes on the surface before and after acid treatment. Alternate soaking approach applied on 10Ce-TZP/Al(2)O(3) consisted of soaking the composite substrates in CaCl(2) and Na(2)HPO(4) solutions alternately to make a precursor for apatite formation. The bioactivity was evaluated by apatite-forming ability of surface-treated materials in simulated body fluid (SBF). Both methods resulted in the formation of hydroxyapatite on the surface of materials; however, alternate soaking approach showed to be a simpler, faster, and more effective method than the chemical treatment approach for enhancing the bioactivity of zirconia materials.     

Calcium-phosphate surface coating by casting to improve bioactivity of titanium

In order to improve the bioactivity of titanium, an original surface treatment was attempted with the use of a casting technique was attempted. Pure titanium was cast into a special graphite mold in which the cavity wall was coated with hydroxyapatite (HA) powder. According to analyses of X-ray diffraction and EDX, the existence of HA and CaO and uptake of Ca and P on the surface of the titanium castings were identified. By immersing the specimen in Hank's solution, the concentrations of Ca and P on the surface increased with immersion time, and the formation of a thin layer with characteristics of spherical HA precipitates was observed after 1 week. The concentrations of Ca and P elements and the Ca/P ratio on the HA layer increased with immersion time. The formation of the HA layer on the titanium cast by this treatment was significantly accelerated compared with pure titanium. The present surface treatment of Ti is expected to improve early bone fixation of Ti implants.     

Calcified nanostructured silicon wafer surfaces for biosensing: effects of surface modification on bioactivity

The growth of known biologically-relevant mineral phases on semiconducting surfaces is one strategy to explicitly induce bioactivity in such materials, either for sensing or drug delivery applications. In this work, we describe the use of a spark ablation process to fabricate deliberate patterns of Ca(10)(PO4)6(OH)2 on crystalline Si (calcified nanoporous silicon). These patterns have been principally characterized by scanning electron microscopy in conjunction with elemental characterization by energy dispersive x-ray analysis. This is followed by a detailed comparison of the effects of fibroblast adhesion and proliferation onto calcified nanoporous Si, calcified nanoporous Si derivatized with alendronate, as well as control samples of an identical surface area containing porous SiO2. Fibroblast adhesion and proliferation assays demonstrate that a higher density of cells grow on the Ca3(PO4)2/porous Si/SiO2 structures relative to the alendronate-modified surfaces and porous Si/SiO2 samples.     

钛表面微弧氧化涂层的细胞生物活性

背景：微弧氧化技术可改善钛或钛合金的表面特征。 目的：研究纯钛表面微弧氧化涂层的表面性能及其对MC3T3-E1细胞早期黏附、增殖及成骨能力的影响。 方法：将46个直径10 mm、厚度2 mm圆盘状纯钛试件分为实验组和对照组。实验组置于含0.02 mol/Lβ-甘油磷酸二钠盐及0.2 mol/L乙酸钙的电解液中进行微弧氧化处理,对照组对试件进行机械抛光。扫描电子显微镜观察试件表面形貌,X射线能谱分析检测涂层表面钙磷比,X射线衍射分析检测涂层晶相构成。将MC3T3-E1细胞接种在两组试件表面,1,2,4 h电镜下观察细胞形态,在2,4,7 d通过CCK-8方法检测细胞增殖,并于7,14 d检测碱性磷酸酶活性。 结果与结论：经微弧氧化处理后,钛表面形成粗糙多孔的钙磷涂层,微弧氧化涂层主要元素为Ca、P、O及Ti,微弧氧化膜层主要由氧化钛、钛酸钙、磷酸钙及偏磷酸钙构成。电镜观察显示1 h 微弧氧化涂层表面细胞已伸出伪足,4 h呈现较典型的细胞形态。细胞在微弧氧化处理钛表面4,7 d的细胞增殖和7,14 d的碱性磷酸酶活性高于对照组。表明微弧氧化技术生成的粗糙多孔钙磷涂层能显著促进MC3T3-E1细胞的早期黏附、增殖及成骨活性。     

Cytocompatibility,osseointegration, and bioactivity of three-dimensional porous and nanostructured network on polyetheretherketone

Porous biomaterials with the proper three-dimensional (3D) surface network can enhance biological functionalities especially in tissue engineering, but it has been difficult to accomplish this on an important biopolymer, polyetheretherketone (PEEK), due to its inherent chemical inertness. In this study, a 3D porous and nanostructured network with bio-functional groups is produced on PEEK by sulfonation and subsequent water immersion. Two kinds of sulfonation-treated PEEK (SPEEK) samples, SPEEK-W (water immersion and rinsing after sulfonation) and SPEEK-WA (SPEEK-W with further acetone rinsing) are prepared. The surface characteristics, in vitro cellular behavior, in vivo osseointegration, and apatite-forming ability are systematically investigated by X-ray photoelectron spectroscopy, Fourier-transform infrared spectroscopy, scanning electron microscopy, cell adhesion and cell proliferation assay, real-time RT-PCR analysis, micro-CT evaluation, push-out tests, and immersion tests. SPEEK-WA induces pre-osteoblast functions including initial cell adhesion, proliferation, and osteogenic differentiation in vitro as well as substantially enhanced osseointegration and bone-implant bonding strength in vivo and apatite-forming ability. Although SPEEK-W has a similar surface morphology and chemical composition as SPEEK-WA, its cytocompatibility is inferior due to residual sulfuric acid. Our results reveal that the pre-osteoblast functions, bone growth, and apatite formation on the SPEEK surfaces are affected by many factors, including positive effects introduced by the 3D porous structure and SO₃H groups as well as negative ones due to the low pH environment. Surface functionalization broadens the use of PEEK in orthopedic implants.     

Fluoroapatite glass-ceramic coating on alumina: surface behavior with biological fluids

The results of a surface analysis performed on a fluoroapatite-based glass ceramic (SAF) also coating a full-density alpha-alumina substrate (SAF-alumina coating) are presented. These two materials have also been evaluated after soaking in simulated body fluid to understand their ability to induce hydroxyapatite growth on them. Aiming to understand the fluoroapatite glass-ceramic interaction with some plasma proteins, in the second part of this study, fibronectin, albumin, immunoglobulin G, IgA, and complement factor C3c SAF binding have been evaluated; surface activity on complement activation has also been quantified. SAF-alumina coating provides good sites for the nucleation and growth of an apatite layer, equivalent to the mineral component of bone and binds preferentially plasma fibronectin, which is well known to enhance cell adhesion and spreading. Moreover, SAF-alumina coating reduces alumina complement activation directly or via reduced IgA binding. Alumina was shown to bind the same C3 fragments as Zymosan, used as complement activating control, and to induce increased levels of serum soluble iC3b and Bb. A mechanical resistant material with enhanced bioactivity, bone integration, and reduced inflammatory potential respect to alumina has been obtained.     

Bioactivity and osteoblast response of the micro-arc oxidized zirconia films

Zirconia films containing Ca and P were prepared by micro-arc oxidation (MAO) of zirconium. The microstructure, in vitro bioactivity, and primary osteoblast response of the films were investigated as a function of the applied voltages in the range of 400-500 V. The results indicate that the MAO-formed zirconia films are porous and nanocrystalline, and predominantly composed of tetragonal zirconia (t-ZrO(2)). The pores and grains sizes and t-ZrO(2) content of the films tend to increase with the applied voltages. The zirconia films formed at higher voltages have higher amount of CaO and phosphate and slightly lower amount of Zr-OH groups. Although, all of the zirconia films can be fully covered by bone-like apatite after immersion in simulated body fluids (SBF) within 10 days, there exists remarkable difference in apatite-induced time. The apatite-forming ability of the films is not only ascribed to Zr-OH groups on the surfaces, but also enhanced by the CaO and phosphate ions incorporated into ZrO(2). Osteoblasts on the films are observed to attach, proliferate, and grow in good state, and have good alkaline phosphatase activity. It is suggested that the MAO-formed ZrO(2) films exhibit favorable bioactivity and biocompatibility.     

Effects of surface undulations of biphasic calcium phosphate tablets on human osteoblast behavior

In this work, the in vitro behavior of human osteoblast cells on the undulated surfaces of biphasic calcium phosphate tablets was investigated. The tablets were produced by uniaxial pressing with convex cylindrical undulations occupying only half of the surface area; the other half was flat. Chemical and physical characterization was performed by scanning electron microscopy (SEM), atomic force microscopy (AFM), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR). XRD and FTIR analyses revealed the presence of hydroxyapatite (HA) and alpha-tricalcium phosphate (alpha-TCP) in a well-defined ratio. Moreover, microtopography, evaluated by SEM and AFM, was similar on the flat region and on that with undulations. However, surface undulations induced different cellular arrangements, confirming the influence of the macrotopography on the cells orientation.     

成骨细胞与镁合金表面硅酸盐-磷酸盐复合涂层的体外相容性

目的 评价成骨细胞在镁合金表面硅酸盐磷酸盐复合涂层上的生长状况,探讨镁合金表面硅酸盐磷酸盐复合涂层与成骨细胞的生物相容性。 方法 用胶原酶消化法分离培养大鼠成骨细胞,并用免疫荧光和茜素红染色法进行鉴定;然后将第4代大鼠的成骨细胞接种于材料表面,用扫描电镜观察成骨细胞在不同表面上黏附形态的变化,同时制备材料浸提液,用浸提液法培养细胞,并通过四甲基偶氮唑盐（MTT）法和碱性磷酸酶（ALP）活性法检测成骨细胞的增殖和分化情况。 结果 将成骨细胞与材料进行复合培养后,扫描电镜下可见,涂层组的成骨细胞生长活力旺盛、形态饱满、分布均匀,而单纯镁合金组未见细胞生长;MTT和ALP活性分析结果显示,硅含量为1%~5%涂层对大鼠成骨细胞的体外生长、增殖和分化的促进作用最佳。 结论 镁合金表面硅酸盐磷酸盐涂层与成骨细胞具有良好的相容性,在体外培养的环境下,有利于细胞的生长、黏附、增殖和分化。     

Reinforcement of bone cement using zirconia fibers with and without acrylic coating

Acrylic (polymethylmethacrylate or PMMA) bone cement was modified by the addition of high-strength zirconia fibers with average lengths of 200 microm and diameters of 15 microm or 30 microm. A novel emulsion polymerization process was developed to encapsulate individual fibers in PMMA. Improvements in tensile and compressive properties as well as in fracture toughness were investigated upon incorporation of uncoated and acrylic coated zirconia fibers. Bone cements were reinforced with 2% by volume of the 15 microm diameter and 5% by volume of the 30 microm fibers. Results indicate that elastic modulus and ultimate strength of bone cements reinforced with zirconia fibers were higher than controls, being the largest for cements reinforced with 30 microm diameter fibers. The fracture toughness of the cement increased by 23% and 41% by the addition of 15 microm and 30 microm fibers, respectively. Coating of individual zirconia fibers did not result in improved material properties of bone cements. The use of uncoated or acrylic coated 30 microm fibers is recommended based on the significant increases in ultimate strength and fracture toughness of the cements.     

Increased osteoblast adhesion on nanograined hydroxyapatite and partially stabilized zirconia composites

To improve the mechanical properties of hydroxyapatite (HA, Ca(10)(PO(4))(6)(OH)(2)) for orthopedic applications, numerous investigators have proposed combining HA with high strength materials, specifically zirconia. Despite the fact that, compared to pure HA, it is now well-established that zirconia and HA composites have improved mechanical properties, the cytocompatibility properties of this composite remain largely uninvestigated. For these reasons, the objective of the present in vitro study was to synthesize HA and partially stabilized zirconia composites for osteoblast (bone-forming cell) adhesion assays. Various sintering temperatures and amounts of zirconia in HA composites were used in order to ascertain their influence on osteoblast adhesion. Results demonstrated increased interactions between HA and partially stabilized zirconia, when either higher sintering temperatures (between 900 and 1,300 degrees C for 1 h) or higher zirconia contents (between 10 and 40 wt %) were used during material synthesis. More importantly, greater osteoblast adhesion was measured on HA-zirconia composites sintered either at lower temperatures (specifically, 900 degrees C) or with lower amounts of zirconia added to HA composites (specifically, 10 wt %). Results further indicated that when sintered at lower temperatures the composites possessed smaller nanometer grain sizes with increased surface roughness and a more stable HA phase. For these reasons, this study suggests that to optimize osteoblast adhesion on HA and partially stabilized zirconia composites for orthopedic applications, low sintering temperatures and low amounts of zirconia should be used. This suggests that a delicate balance must be reached between increasing mechanical properties of HA without decreasing osteoblast cytocompatibility properties through zirconia addition.     

In vitro bioactivity and phase stability of plasma-sprayed nanostructured 3Y-TZP coatings

In this work, plasma-sprayed nanostructured zirconia coatings stabilized with 3 mol.% yttria (3Y-TZP) were deposited on Ti substrates. The microstructure and phase composition of coatings were characterized using scanning electron microscopy and X-ray diffraction. The in vitro bioactivity of coatings was evaluated by examining the formation of bone-like apatite on its surface in simulated body fluid. MG63 cell lines were cultured on the coating to investigate its cytocompatibility. The crystalline phase of the as-sprayed coating was tetragonal zirconia, and no monoclinic zirconia was detected. The size of the grains on the as-sprayed coating surface was less than 100 nm. The apatite could precipitate on the surface of the coating immersed in simulated body fluid for 28 days while no apatite was formed on the surface of 3Y-TZP ceramic control, indicating that the bioactivity of the coating is superior to the ceramic with the same composition. It also revealed that the polished coating whose nanostructural outmost layer was removed was bioinert, implying the significance of the nanosized grains for its bioactivity. MG63 cells could adhere, grow and proliferate well on the coating surface, indicating that the coating had good cytocompatibility. Phase stability of plasma-sprayed 3Y-TZP coating was evaluated under hydrothermal conditions at 134 °C. It revealed that the plasma-sprayed nanostructured zirconia coating was more sensitive to aging than that of zirconia ceramics.     

In vitro bioactivity of laser ablation pseudowollastonite coating

Pseudowollastonite (psW) coatings on titanium alloys substrates were prepared by laser ablation and immersed in simulated body fluid (SBF) for different periods in order to investigate the nucleation and growth of hydroxyapatite (HA)-like formation on their surface. The structure of the coatings before soaking was analysed by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The interfacial reactions product was examined by thin-film XRD, SEM and transmission electron microscopy at low and high resolution level, both fitted with energy-dispersive X-ray spectroscopy. Additional changes in ionic concentration, using inductively couple plasma atomic emission spectroscopy, were determined as well as pH right at the psW-coatings/SBF interface using an ion-sensitive field effect transistor. The solution composition changes, increasing the Ca(2+) and Si(4+) concentration and pH as a function of the soaking time while HPO(4)(2-) decreased.The results obtained showed that the coating surfaces were covered by HA-like, which indicated that the psW-coating possesses good bioactivity and also suggested that the mechanism of HA-like layer formation in SBF was similar to that showed in in vitro test by other silica-based materials.     

Synthesis, characterization, bioactivity and biocompatibility of nanostructured materials based on the wollastonite-poly(ethylmethacrylate-co-vinylpyrrolidone) system

Composite materials are very promising biomaterials for hard tissue augmentation. The approach assayed in this work involves the manufacturing of a composite made of a bioactive ceramic, natural wollastonite (W) and a nanostructured copolymer of ethylmethacrylate (EMA) and vinylpyrrolidone (VP) to yield a bioresorbable and biocompatible VP-EMA copolymer. A bulk polymerization was induced thermally at 50 degrees C, using 1 wt % azobis(isobutyronitrile) (AIBN) as free-radical initiator. Structural characterization, compressive strength, flexural strength (FS), degradation, bioactivity, and biocompatibility were evaluated in specimens with a 60/40 VP/EMA ratio and ceramic content in the range 0-60%. A good integration between phases was achieved. Greater compression and FS, in comparison with the pure copolymer specimens was obtained only when the ceramic load got up to 60% of the total weight. The soaking in NaCl solution resulted in the initial swelling of the specimens tested. The maximum swelling was reached after 2-3 h of immersion and it was significantly greater for lower ceramic loads. This result makes the polymer component the main responsible for the interactions with the media. After soaking in SBF, microdomains segregation can be observed in the polymer component that can be related with a dramatic difference in the reactivity of both monomers in free radical polymerization, whereas the formation of an apatite-like layer on the W surfaces can be observed. Biocompatibility in vitro studies showed the absence of cytotoxicity of all formulations. The cells were able to adhere on the polystyrene negative control and on specimens containing 60 wt % wollastonite forming a monolayer and showing a normal morphology. However, a low cellular growth was observed.     

Low-temperature degradation in zirconia with a porous surface

Today there is growing interest in zirconia in the dental field, but its use is still recent. Dental zirconia is mainly found in the form of yttria-stabilized zirconia crowns, bridges and abutments, and several companies are developing zirconia implants as an alternative to the standard biomedical grade titanium. In order to favor bone in-growth and osseointegration of zirconia implants, several strategies are now being explored to process rough and/or porous surfaces. The aim of this paper was to evaluate the resistance to environmental degradation of yttria-stabilized zirconia coated with a porous layer. We show that specific conditions of processing to generate the porous layer at the surface can lead to an accelerated tetragonal-monoclinic transformation of the porous layer in the presence of water. The impact of the transformation was evaluated in terms of structural integrity. Bending strength was not affected but the cohesion of the porous coating and its adhesion with the dense part deteriorated. We show that other processing conditions insure much better stability. Low-temperature degradation resistance of such porous surfaces should therefore be carefully followed and controlled in order to avoid critical problems in the future.     

Influence of recovering collagen with bioactive glass on osteoblast behavior

Bioactive ceramics have interesting properties from the biological standpoint, but their effects on cellular events remain partially unknown. In the current work, we investigated cellular viability, proliferation, and metabolic activity of rat primary osteoblasts in contact with four different samples: type I collagen, bioactive glass-coated collagen (GC), and both samples submitted to immersion for 5 days in a simulated body fluid. The bioactive glass coating was obtained from a sol-gel process. The cell viability, the alkaline phosphate, the collagen secretion, and the nitric oxide production by osteoblast were measured after 72 h of incubation in the presence of the samples. The GC that was immersed for 5 days in a simulated body fluid solution showed an increase in osteoblast viability and proliferation when it was compared with control and the other samples.     

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.     

In vitro bioactivity of S520 glass fibers and initial assessment of osteoblast attachment

Bioactive glass fibers are attractive materials for use as tissue-engineering scaffolds and as the reinforcing phase for resorbable bioactive composites. The bioactivity of S520 glass fibers (52.0 mol % SiO(2), 20.9 Na(2)O, 7.1 K(2)O, 18.0 CaO, and 2.0 P(2)O(5)) was evaluated in two media, simulated body fluid (SBF) and Dulbecco's modified Eagle's medium (DMEM), for up to 20 days at 37 degrees C. Hydroxyapatite formation was observed on S520 fiber surfaces after 5 h in SBF. After a 20-day immersion, a continuous hydroxyapatite layer was present on the surface of samples immersed in SBF as well as on those samples immersed in DMEM [fiber surface area to solution volume ratio (SA:V) of 0.10 cm(2)/mL]. Backscattered electron imaging and EDS analysis revealed that the hydroxyapatite layer formation was more extensive for samples immersed in SBF. Decreasing the SA:V ratio to 0.05 cm(2)/mL decreased the time required to form a continuous hydroxyapatite surface layer. ICP was used to reveal Si, Ca, and P release profiles in DMEM after the 1st h (15.1, 83.8, and 29.7 ppm, respectively) were similar to those concentrations previously determined to stimulate gene expression in osteoblasts in vitro (16.5, 83.3, and 30.4 ppm, respectively). The tensile strength of the 20-microm diameter fibers was 925 +/- 424 MPa. Primary human osteoblast attachment to the fiber surface was studied by using SEM, and mineralization was studied by using alizarin red staining. Osteoblast dorsal ruffles, cell projections, and lamellipodia were observed, and by 7 days, cells had proliferated to form monolayer areas as shown by SEM. At 14 days, nodule formation was observed, and these nodules stained positive for alizarin red, demonstrating Ca deposition and, therefore mineralization.     

骨用镁合金表面Ti-Cu涂层的腐蚀行为

背景：镁合金作为骨折内固定材料具有力学性能与骨相近、生物相容性好和可降解等突出优点,但其在体液中降解速度过快,成为其临床应用的瓶颈,因此综合利用表面处理提高其耐蚀性能具有重要意义。 目的：综合运用磁控溅射技术和碱热处理技术在镁合金表面制备兼具耐蚀性能和生物活性的涂层。 方法：首先采用熔炼技术制备Mg-Zn-Mn合金,利用磁控溅射技术在材料表面制备致密涂层,然后利用碱性溶液对表面涂层进行处理,利用模拟体液浸泡实验研究涂层的腐蚀行为,通过表面产物中钙和磷的含量推测涂层的生物活性。 结果与结论：经磁控溅射和碱热处理技术在镁合金表面制备兼具耐蚀性能和生物活性的涂层;经模拟体液浸泡24和168 h后涂层表面沉积含Ca,P产物,Ca/P比分别为1.54和2.11,接近类骨磷酸盐Ca/P比,涂层表面的浸泡24 h形成5-10 μm点腐蚀,随着浸泡时间增加点腐蚀逐渐长大,浸泡168 h后点腐蚀增加为100-800 μm。     

Microstructure and mechanical properties of hydroxyapatite ceramics with zirconia dispersion prepared by post-sintering

Hydroxyapatite ceramics with zirconia dispersion from fine powders synthesized hydrothermally were post-sintered at 1000-1300 degrees C under 200 MPa of argon for 1 h without capsules, after normal sintering in air at 1200 degrees C for 3 h. Densification was most significant with post-sintering at 1200 degrees C. Fracture toughness, Vickers hardness and elastic properties of these materials were investigated. Post-sintering gave twice the K1c value of transparent pure hydroxyapatite ceramics. Vickers hardness and Young's modulus of the ceramics were increased by post-sintering.