Functions of miR-1 and miR-133a during the postnatal development of masseter and gastrocnemius muscles.

The present study investigated the function of miR-1 and miR-133a during the postnatal development of mouse skeletal muscles. The amounts of miR-1 and miR-133a were measured in mouse masseter and gastrocnemius muscles between 1 and 12 weeks after birth with real-time polymerase chain reaction and those of HDACs, MEF2, MyoD family, MCK, SRF, and Cyclin D1 were measured at 2 and 12 weeks with Western blotting. In both the masseter and gastrocnemius muscles, the amount of miR-1 increased between 1 and 12 weeks, whereas the amount of HADC4 decreased between 2 and 12 weeks. In the masseter muscle, those of MEF2, MyoD, Myogenin, and MCK increased between 2 and 12 weeks, whereas, in the gastrocnemius muscle, only those of MRF4 and MCK increased. The extent of these changes in the masseter muscle was greater than that in the gastrocnemius muscle. The amounts of miR-133a, SRF, and Cyclin D1 did not change significantly in the masseter muscle between 1 and 12 weeks after birth. By contrast, in the gastrocnemius muscle, the amounts of miR-133a and Cyclin D1 increased, whereas that of SRF decreased. Our findings suggest that the regulatory pathway of miR-1 via HDAC4 and MEF2 plays a more prominent role during postnatal development in the masseter muscle than in the gastrocnemius muscle, whereas that of miR-133a via SRF plays a more prominent role in the gastrocnemius muscle than in the masseter muscle.

Seasonal and environmental changes of mycorrhizal associations and heterotrophy levels in mixotrophic Pyrola japonica (Ericaceae) growing under different light environments.

PREMISE OF THE STUDY: Mixotrophy is a strategy whereby plants acquire carbon both through photosynthesis and heterotrophic exploitation of mycorrhizal fungi. In Euro-American Pyroleae species studied hitherto, heterotrophy levels vary according to species, sites of study, and possibly light conditions. We investigated mycorrhizal association and mixotrophy in the Asiatic forest species Pyrola japonica, and their plasticity under different light conditions. METHODS: Pyrola japonica was sampled bimonthly in sunny and shaded conditions from a deciduous broadleaf forest. We microscopically assessed the rate of fungal colonization and sequenced the ITS to identify the mycorrhizal fungi. We measured (13)C and (15)N isotopic abundances in P. japonica as compared with neighboring autotrophic and mycoheterotrophic plants, to evaluate P. japonica's heterotrophy level. KEY RESULTS: Pyrola japonica formed arbutoid mycorrhizas devoid of fungal mantles, with intracellular hyphal coils and a Hartig net. It tended to be more colonized by mycorrhizal fungi in spring and summer. Most associated fungi belonged to ectomycorrhizal taxa, and 84% of identified fungi were Russula spp. Rate of mycorrhizal colonization and Russula frequency tended to be higher in shaded conditions. Both δ(13)C and δ(15)N values of P. japonica were significantly higher in autotrophic plants, showing that about half of the carbon on average was received from mycorrhizal fungi. Both isotopic values negatively correlated with light availability, suggesting higher heterotrophy levels in shaded conditions. CONCLUSIONS: The mixotrophic P. japonica undergoes changes in mycorrhizal symbionts and carbon nutrition according to light availability. Our results suggest that during Pyroleae evolution, a tendency to increased heterotrophy emerged in the Pyrola/Orthilia clade.

The significance of thermomechanical processing on the cellular response of biomedical Co-Cr-Mo alloys.

Strengthening of biomedical Co-Cr-Mo alloys has been explored via thermomechanical processing for enhancing the durability of their biomedical applications. However, the effects of cold and hot deformation on the cellular activity continue to be unclear. In this study, we prepared Co-Cr-Mo alloy rods via cold swaging and hot-caliber rolling and studied the relationship between the microstructure and cellular response of pre-osteoblasts. The cold-swaged rod experienced strain-induced martensitic transformation, which increased the volume fraction of the hexagonal close-packed (hcp) ε-martensite to ∼60 vol.% with an increase in area reduction (r) to 30%. The 111γ fiber texture of the face-centered cubic (fcc) γ-matrix followed the Shoji-Nishiyama orientation relationship with ε-martensite. Cell culture results revealed beneficial effects of cold swaging on the cell response, in terms of adhesion, proliferation and morphology of cells, although increasing r did not significantly affect cellular metabolism levels. The addition of small content of Zr (0.04 wt.%) led to enhanced focal adhesion of cells, which became more significant at higher r. The microstructural evolution during hot-caliber rolling, namely, grain refinement without any phase transformation and strong texture development, did not appreciably affect the cellular activity. These findings are envisaged to facilitate alloy design and microstructural optimization for favorable tuning the osseointegration of biomedical Co-Cr-Mo alloys.

Favorable modulation of osteoblast cellular activity on Zr-modified Co-Cr-Mo alloy: The significant impact of zirconium on cell-substrate interactions.

Cobalt-chromium-molybdenum alloys exhibit good mechanical properties (yield strength: ~530 MPa, ultimate tensile strength: ~1114 MPa, elongation-to-failure: ~47.3%, and modulus: ~227 GPa) and corrosion resistance. In recent years, from the perspective of osseointegration, they are considered to be lower in rank in comparison to the widely used titanium alloys. We elucidate here the significant and favorable modulation of cellular activity of Zr-modified Co-Cr-Mo alloys. The average grain size of Co-Cr-Mo alloy samples with and without Zr was 104 ± 27 and ~53 ± 11 µm, respectively. The determining role of small addition of Zr (0.04 wt. %) to the Co-Cr-Mo alloys in favorable modulation of cellular activity was accomplished by combining cellular biology and materials science and engineering. Experiments on the influence of Zr addition to Co-Cr-Mo alloys clearly demonstrated that the cell adhesion, spread and cell-substrate interactions were enhanced in the presence of Zr. The spread/growth rate of cells was ~120% on the Co-Cr-Mo alloy and 190% per day on the Co-Cr-Mo-Zr alloy. While the % area covered by the cells increased from ~5.1 to ~33.6% on Co-Cr-Mo alloy and ~19.2 to ~47.8% on Co-Cr-Mo-Zr alloy after 2 and 24 hr of incubation. Similarly, the cell density increased from ~1354 to ~3424 cells/cm2 on Co-Cr-Mo alloy and ~3583 to ~7804 cells/cm2 on Co-Cr-Mo-Zr alloy after 2 and 24 hr of incubation. Additionally, stronger vinculin focal adhesion contact and signals associated with actin stress fibers together with extracellular matrix protein, fibronectin, were noted.

Tuning strain-induced γ-to-ε martensitic transformation of biomedical Co-Cr-Mo alloys by introducing parent phase lattice defects.

In this study, we examined the effect of pre-existing dislocation structures in a face-centered cubic γ-phase on strain-induced martensitic transformation (SIMT) to produce a hexagonal close-packed ε-phase in a hot-rolled biomedical Co-Cr-Mo alloy. The as-rolled microstructure was characterized by numerous dislocations as well as stacking faults and deformation twins. SIMT occurred just after macroscopic yielding in tensile deformation. Using synchrotron X-ray diffraction line-profile analysis, we successfully captured the nucleation of ε-martensite during tensile deformation in terms of structural evolution in the surrounding γ-matrix: many dislocations that were introduced into the γ-matrix during the hot-rolling process were consumed to produce ε-martensite, together with strong interactions between dislocations in the γ-matrix. As a result, the SIMT behavior during tensile deformation was accelerated through the consumption of these lattice defects, and the nucleation sites for the SIMT ε-phase transformed into intergranular regions upon hot rolling. Consequently, the hot-rolled Co-Cr-Mo alloy simultaneously exhibited an enhanced strain hardening and a high yield strength. The results of this study suggest the possibility of a novel approach for controlling the γ → ε SIMT behavior, and ultimately, the performance of the alloy in service by manipulating the initial dislocation structures.

Effect of nitrogen on the microstructure and mechanical properties of Co-33Cr-9W alloys prepared by dental casting.

The effect of nitrogen concentration on the mechanical properties of Co-33Cr-9W alloy dental castings fabricated using the "high-Cr and high-N" concept was investigated. Microstructural analysis was performed on the alloys, and findings were discussed in relation to the mechanical properties. Owing to their high nitrogen concentrations (0.25-0.35wt%), all alloys prepared exhibited face-centered cubic (fcc) γ-phase matrices with a-few-millimeter grains consisting of dendritic substructures. Strain-induced martensitic transformations to produce hexagonal close-packed (hcp) ε-phases were not identified under tensile deformation. The precipitation of the intermetallic σ-phase was identified at the interdendritic regions where solidification segregation of Cr and W occurred. The size and chemical composition of this σ-phase did not vary with the bulk nitrogen concentration. Adding nitrogen to the alloys did not alter their tensile yield stress or Vickers hardness values significantly, suggesting that the nitrogen strengthening effect is affected by the manufacturing route as well as local chemistry that is involved in the microstructural evolution during solidification. The tensile ductility, on the other hand, increased with an increase in nitrogen concentration; the alloy with 0.35wt% nitrogen exhibited 21% elongation with a high 0.2% proof stress (589MPa). This significant improvement in ductility was likely caused by the reduction in the amount of σ-phase precipitates at the interdendritic regions following the addition of nitrogen.

Impact of minor alloying with C and Si on the precipitation behavior and mechanical properties of N-doped Co-Cr alloy dental castings.

The addition of carbon and silicon as minor alloying elements was examined as a means to improve the mechanical properties of novel nitrogen-doped Co-Cr-based alloy dental castings. Samples of Co-32Cr-9W-Si-0.25N-C (mass%) alloys were prepared using a dental-casting machine. Microstructural analysis was performed on the alloys using scanning electron microscopy, electron-backscatter diffraction, electron-probe microanalysis, and X-ray diffraction, with a particular focus on the precipitation behavior. The findings were compared with thermodynamic predictions and examined in relation to the tensile properties and Vickers hardness at room temperature. All of the prepared alloys had a face-centered-cubic γ-phase matrix, with grains measuring a few millimeters in diameter and consisting of dendritic substructures. The precipitation of the intermetallic σ-phase, which occurred in the interdendritic regions with solidification segregation of Cr and W, was replaced with M23C6 through the addition of carbon. This significantly increased the ultimate tensile strength of the alloys without severe loss of ductility, although the 0.2% proof stress did not change. The addition of silicon, on the contrary, promoted the formation of the precipitates, which included M6C and the σ-phase, making the alloys brittle. The results of this study highlight the role of minor alloying elements, such as carbon and silicon, on the microstructural and mechanical properties; the findings also shed light on the significance of precipitation control in dental castings of Co-Cr alloys, which should aid the design of novel dental alloys.

Stacking-fault strengthening of biomedical Co-Cr-Mo alloy via multipass thermomechanical processing.

The strengthening of metallic biomaterials, such as Co-Cr-Mo and titanium alloys, is of crucial importance to the improvement of the durability of orthopedic implants. In the present study, we successfully developed a face-centered cubic (fcc) Co-Cr-Mo alloy with an extremely high yield strength (1400 MPa) and good ductility (12%) by multipass hot-rolling, which is suitable for industrial production, and examined the relevant strengthening mechanisms. Using an X-ray diffraction line-profile analysis, we revealed that a substantial increase in the number of stacking faults (SFs) in the fcc γ-matrix occurred at a greater height reduction (r), while physical modeling demonstrated that the contribution of the accumulated SFs (i.e., the reduction in SF spacing) with an increase in r successfully explains the entire strengthening behavior of the hot-rolled alloy. The present study sheds light on the importance of the SF strengthening mechanism, and will help to guide the design and manufacturing strategy for the high-strength Co-Cr-Mo alloys used in highly durable medical devices.

Developing high strength and ductility in biomedical Co-Cr cast alloys by simultaneous doping with nitrogen and carbon.

There is a strong demand for biomedical Co-Cr-based cast alloys with enhanced mechanical properties for use in dental applications. We present a design strategy for development of Co-Cr-based cast alloys with very high strength, comparable to that of wrought Co-Cr alloys, without loss of ductility. The strategy consists of simultaneous doping of nitrogen and carbon, accompanied by increasing of the Cr content to increase the nitrogen solubility. The strategy was verified by preparing Co-33Cr-9W-0.35N-(0.01-0.31)C (mass%) alloys. We determined the carbon concentration dependence of the microstructures and their mechanical properties. Metal ion release of the alloys in an aqueous solution of 0.6% sodium chloride (NaCl) and 1% lactic acid was also evaluated to ensure their corrosion resistance. As a result of the nitrogen doping, the formation of a brittle σ-phase, a chromium-rich intermetallic compound, was significantly suppressed. Adding carbon to the alloys resulted in finer-grained microstructures and carbide precipitation; accordingly, the strength increased with increasing carbon concentration. The tensile ductility, on the other hand, increased with increasing carbon concentration only up to a point, reaching a maximum at a carbon concentration of ∼0.1mass% and decreasing with further carbon doping. However, the alloy with 0.31mass% of carbon exhibited 14% elongation and also possessed very high strength (725MPa in 0.2% proof stress). The addition of carbon did not significantly degrade the corrosion resistance. The results show that our strategy realizes a novel high-strength Co-Cr-based cast alloy that can be produced for advanced dental applications using a conventional casting procedure. STATEMENT OF SIGNIFICANCE: The present study suggested a novel alloy design concept for realizing high-strength Co-Cr-based cast alloys. The proposed strategy is beneficial from the practical point of view because it uses conventional casting approach-a simpler, more cost-effective, industrially friendly manufacturing process than other manufacturing processes such as thermomechanical processing or powder metallurgy. The developed alloys showed the excellent strength-ductility balance and significantly high strength comparable to that of wrought Co-Cr-Mo alloys, while maintaining acceptable ductility and good corrosion resistance. We described the relationship between microstructures and mechanical and corrosion prosperities of the developed alloys; this provides the fundamental aspect of the proposed strategy and will be helpful for further investigations or industrial realization of the proposed strategy.

Characterisation of nanoscale carbide precipitation in as-cast Co-Cr-W-based dental alloys.

Carbide precipitation in biomedical Co-Cr alloys significantly influences the performance in terms of mechanical properties and corrosion resistance. In this study, we examined the carbide precipitation associated with local solute partitioning in as-cast Co-Cr-W-based dental alloys at the micro- and nano-scale. Co-28Cr-9W-1Si (mass%) alloys with carbon concentrations ranging from 0.01 to 0.33 mass% were prepared. It was found that Cr, W, Si, and C segregated at the interdendritic regions of the face-centered cubic (fcc) γ-grain microstructures, resulting in precipitation; the amount of interdendritic precipitates was found to increase with increasing bulk carbon concentration. We identified, for the first time, the nanosized mixed-phase constituents, which were composed of ∼100 nm fine grains of the γ-phase and M23C6-type carbide, at the interdendritic regions of the high-carbon Co-Cr-W alloy. These nanosized M23C6 carbides were produced with a cube-on-cube orientation relationship ([001]γ||[001]M23C6 and (100)γ||(100)M23C6 ) with the surrounding submicron γ-grains, while the local morphology changed from equiaxed to plate-like morphology. Nanoscale partitioning of the alloying elements was considered to be the origin of the interdendritic nanostructured constituents. As such, the nanoscale elemental partitioning was observed to alter the local mechanical properties and to affect the corrosion resistance of the alloys. The observed results indicate the importance of local chemistry-which has not received sufficient attention to date-along with carbide precipitation in the optimization of alloy design for achieving desirable properties such as high strength, ductility, and corrosion resistance in dental Co-Cr-based cast alloys.

Preventing high-temperature oxidation of Co-Cr-based dental alloys by boron doping.

Biomedical Co-Cr-based alloys used in dental restorations are usually subjected to high-temperature treatments during manufacturing. Therefore, it is practically essential to characterise and control the oxide films formed on the surfaces of these alloys during heat treatment in terms of material loss, the accuracy of fit, and the aesthetics of dental restorations. In this work, the effects of boron doping on the surface oxide films formed on Ni-free Co-28Cr-9W-1Si (mass%) dental alloys under short-term exposure to high temperatures, which simulate the manufacturing process of porcelain-fused-to-metal (PFM) restorations, were investigated. The surface oxides primarily consisted of Cr2O3 in all prepared alloys. The chemical composition of these surface layers varies with the B concentration in the bulk, with the addition of boron stabilising the dense Cr2O3 phase in the oxide films. Nanoscale boron enrichment is clearly observed at the interface between the oxide films and the metal substrate, with the oxidation of boron atoms leading to the formation of a B2O3 layer. Since B2O3 and Cr2O3 prevent oxygen diffusion, the surface oxide films on the boron-containing alloys are thinner; however, no additional thinning was observed when increasing the boron content from 0.01 to 0.8 mass%. It was also found that a small amount of boron does not degrade the corrosion properties of the alloys in a 0.9% NaCl solution. The results obtained in this study will aid in the improvement of manufacturing processes, and ultimately, the performance of PFM restorations.

Manufacturing of high-strength Ni-free Co-Cr-Mo alloy rods via cold swaging.

The strengthening of biomedical metallic materials is crucial to increasing component durability in biomedical applications. In this study, we employ cold swaging as a strengthening method for Ni-free Co-Cr-Mo alloy rods and examine its effect on the resultant microstructures and mechanical properties. N is added to the alloy to improve the cold deformability, and a maximum reduction in area (r) of 42.6% is successfully obtained via cold swaging. The rod strength and ductility increase and decrease, respectively, with increasing cold-swaging reduction r. Further, the 0.2% proof stress at r=42.6% eventually reaches 1900MPa, which is superior to that obtained for the other strengthening methods proposed to date. Such significant strengthening resulting from the cold-swaging process may be derived from extremely large work hardening due to a strain-induced γ (fcc)→ε (hcp) martensitic transformation, with the resultant intersecting ε-martensite plates causing local strain accumulation at the interfaces. The lattice defects (dislocations/stacking faults) inside the ε phase also likely contribute to the overall strength. However, excessive application of strain during the cold-swaging process results in a severe loss in ductility. The feasibility of cold swaging for the manufacture of high-strength Co-Cr-Mo alloy rods is discussed.

Development of microstructure and mechanical properties during annealing of a cold-swaged Co-Cr-Mo alloy rod.

In this study, we investigated the evolution of the microstructure and mechanical properties during annealing of a cold-swaged Ni-free Co-Cr-Mo alloy for biomedical applications. A Co-28Cr-6Mo-0.14N-0.05C (mass%) alloy rod was processed by cold swaging, with a reduction in area of 27.7%, and then annealed at 1173-1423K for various periods up to 6h. The duplex microstructure of the cold-swaged rod consisted of a face-centered cubic γ-matrix and hexagonal closed-packed ε-martensite developed during cold swaging. This structure transformed nearly completely to the γ-phase after annealing and many annealing twin boundaries were observed as a result of the heat treatment. A small amount of the ε-phase was identified in specimens annealed at 1173K. Growth of the γ-grains occurred with increasing annealing time at temperatures ≥1273K. Interestingly, the grain sizes remained almost unchanged at 1173K and a very fine grain size of approximately 8µm was obtained. The precipitation that occurred during annealing was attributed to the limited grain coarsening during heat treatment. Consequently, the specimens treated at this temperature showed the highest tensile strength and lowest ductility among the specimens prepared. An elongation-to-failure value larger than 30% is sufficient for the proposed applications. The other specimens treated at higher temperatures possessed similar tensile properties and did not show any significant variations with different annealing times. Optimization of the present rod manufacturing process, including cold swaging and interval annealing heat treatment, is discussed.

Cold-rolling behavior of biomedical Ni-free Co-Cr-Mo alloys: Role of strain-induced ε martensite and its intersecting phenomena.

Ni-free Co-Cr-Mo alloys are some of the most difficult-to-work metallic materials used commonly in biomedical applications. Since the difficulty in plastically deforming them limits their use, an in-depth understanding of their plastic deformability is of crucial importance for both academic and practical purposes. In this study, the microstructural evolution of a Co-29Cr-6Mo-0.2N (mass%) alloy during cold rolling was investigated. Further, its work-hardening behavior is discussed while focusing on the strain-induced face-centered cubic (fcc) γ→hexagonal close-packed (hcp) ε martensitic transformation (SIMT). The planar dislocation slip and subsequent SIMT occurred even in the initial stage of the deformation process owing to the low stability of the γ-phase and contributed to the work hardening behavior. However, the amount of the SIMTed ε-phase did not explain the overall variation in work hardening during cold rolling. It was found that the intersecting of the SIMTed ε-plates enhanced local strain evolution and then produced fine domain-like deformation microstructures at the intersections. Consequently, the degree of work hardening was reduced during subsequent plastic deformation, resulting in the alloy exhibiting a two-stage work hardening behavior. The results obtained in this study suggest that the interaction between ε-martensites, and ultimately its relaxation mechanism, is of significant importance; therefore, this aspect should be addressed in detail; the atomic structures of the γ-matrix/ε-martensite interfaces, the phenomenon of slip transfer at the interfaces, and the slipping behavior of the ε-phase itself are needed to be elucidated for further increasing the cold deformability of such alloys.

Assessment of precipitation behavior in dental castings of a Co-Cr-Mo alloy.

This study investigated solute portioning and precipitation in dental castings of a Co-Cr-Mo alloy and discussed their effects on alloy performance, in particular, the mechanical properties. Samples of a commercial Co-29Cr-6Mo (mass%) alloy were prepared using a dental-casting machine. The precipitates formed owing to the partitioning behaviors of the alloying elements were investigated using scanning electron microscopy, electron backscatter diffraction analysis, electron probe microanalysis, and transmission electron microscopy. The prepared samples exhibited a very coarse face-centered-cubic γ-phase dendritic structure with an average grain size of a few millimeters. A large number of precipitates, which decomposed further into complex interdendritic constituents (σ- and M23C6 carbide phases) were observed in the interdendritic regions rich in Cr, Mo, Si, and C. A reaction between the σ-phase and carbon is probably responsible for the carbide M23C6; however, this reaction did not occur to completion in the current case in spite of slow cooling (i.e., long exposure to elevated temperatures) in dental casting. While these precipitates result in high strength (hardness) and/or brittleness, the properties can be improved further by optimizing the alloy composition and the manufacturing process. The results of this study shed light on the significance of precipitation control in dental castings of Co-Cr-Mo alloys and should aid in the design of novel biomedical Co-Cr-based dental alloys that exhibit better performances.

Texture evolution and mechanical anisotropy of biomedical hot-rolled Co-Cr-Mo alloy.

Crystallographic textures and their effect on the mechanical anisotropy of a hot-rolled biomedical Co-Cr-Mo alloy were investigated. The hot-rolled Co-28Cr-6Mo-0.13N (mass%) alloy examined here exhibited a monotonic strength increment following hot-rolling reduction, eventually reaching a 0.2% proof stress of 1400 MPa while maintaining acceptable ductility (>10%). The dominant hot-rolling texture was a brass-type component, which is characterized by the alloy's peculiarly low stacking fault energy (SFE) even at hot rolling temperatures, although the minor peaks of the near copper component were also identified. However, because of the onset of dynamic recrystallization (DRX) during the hot rolling process, the texture intensity was relatively weak even after 90% hot rolling, although the grain refinement originating from the DRX was not significant (the "less active DRX" condition increased the strain accumulation during the process, resulting in high-strength samples). The weakened texture development resulted in negligible in-plane anisotropy for the hot-rolled specimen strength, when the specimens were tensile strained in the rolling direction (RD) and transverse direction (TD). The elongation-to-failure, however, exhibited a difference with respect to the tensile loading axis. It is suggested that the ductility anisotropy is closely related to a strain-induced γ (fcc) → ε (hcp) martensitic transformation during tensile loading, resulting in a difference in the proportion of quasi-cleavage fracture surfaces. The obtained results will be helpful in the development of high-strength Co-Cr-Mo alloy plates and sheets, and have implications regarding plastic deformation and texture evolution during the hot rolling of non-conventional metallic materials with low SFE at elevated temperatures, where planar dislocation slips of Shockley partial dislocations and thermally activated process interplay.

Effect of carbon on the microstructure, mechanical properties and metal ion release of Ni-free Co-Cr-Mo alloys containing nitrogen.

This paper investigated the effect of carbon addition on the microstructure and tensile properties of Ni-free biomedical Co-29Cr-6Mo (mass%) alloys containing 0.2 mass% nitrogen. The release of metal ions by the alloys was preliminarily evaluated in an aqueous solution of 0.6% sodium chloride (NaCl) and 1% lactic acid, after which samples with different carbon contents were subjected to hot rolling. All specimens were found to primarily consist of a γ-phase matrix due to nitrogen doping, with only the volume fraction of M23C6 increasing with carbon concentration. Owing to the very fine size of these carbide particles (less than 1 µm), which results from fragmentation during hot rolling, the increased formation of M23C6 increased the 0.2% proof stress, but reduced the elongation-to-failure. Carbon addition also increased the amount of Co and Cr released during static immersion; Co and Cr concentrations at the surfaces, which increased with increasing the bulk carbon concentrations, possibly enhanced the metal ion release. However, only a very small change in the Mo concentration was noticed in the solution. Therefore, it is not necessarily considered a suitable means of improving the strength of biomedical Co-Cr-Mo alloys, even though it has only to date been used in this alloy system. The results of this study revealed the limitations of the carbon strengthening and can aid in the design of biomedical Co-Cr-Mo-based alloys that exhibit the high durability needed for their practical application.

Analysis of the Fracture Mechanism of Ti-6Al-4V Alloy Rods That Failed Clinically After Spinal Instrumentation Surgery.

STUDY DESIGN: Retrieval analysis of 2 Ti-6Al-4V alloy rods that fractured after spinal instrumentation surgery. OBJECTIVE: To determine the mechanism that underlies fractures of Ti-6Al-4V alloy rods after spinal instrumentation surgery from a materials science viewpoint. SUMMARY OF BACKGROUND DATA: Rod failures after spinal instrumentation surgery are often reported and many case-based studies have been published. However, the details of the mechanism that underlies the fractures have not yet been fully elucidated. METHODS: Two patients, a 71-year-old female and an 11-year-old male, underwent radiography and removal of their fractured rods. The latter patient had been treated using the growing-rod method. Metallurgical failure analysis of the retrieved rods was conducted, and material properties were compared between the unused and fractured rods. RESULTS: The microstructures and mechanical properties of the Ti-6Al-4V alloy rods that failed after spinal instrumentation surgery were similar to those of unused rods. Analysis of the fracture surfaces clearly identified fatigue cracking in both cases that would have lowered the resistance of the rods to failures caused by external stresses. Shot blasting the surfaces of Ti-6Al-4V alloy rods and bending the rods to fit particular contours, which is always conducted during spinal instrumentation surgery, probably introduced fatigue cracking because the alloy is highly notch sensitive. CONCLUSION: Improvements should be made to rod design and/or rod material, because the fatigue resistance of titanium alloys is intrinsically lower than that of other commercially available rod materials, including cobalt-chromium alloys. These imperfections may have greater consequences for the growing-rod method and pseudarthrosis, where the rods are not completely fixed, and they subsequently suffer from severe long-arm moments. LEVEL OF EVIDENCE: N/A.

Strengthening of biomedical Ni-free Co-Cr-Mo alloy by multipass "low-strain-per-pass" thermomechanical processing.

Further strengthening of biomedical Co-Cr-Mo alloys is desired, owing to the demand for improvements to their durability in applications such as artificial hip joints, spinal rods, bone plates, and screws. Here, we present a strategy-multipass "low-strain-per-pass" thermomechanical processing-for achieving high-strength biomedical Co-Cr-Mo alloys with sufficient ductility. The process primarily consists of multipass hot deformation, which involves repeated introduction of relatively small amounts of strain to the alloy at elevated temperatures. The concept was verified by performing hot rolling of a Co-28 Cr-6 Mo-0.13N (mass%) alloy and its strengthening mechanisms were examined. Strength increased monotonically with hot-rolling reduction, eventually reaching 1,400 MPa in 0.2% proof stress, an exceptionally high value. Synchrotron X-ray diffraction (XRD) line-profile analysis revealed a drastic increase in the dislocation density with an increase in hot-rolling reduction and proposed that the significant strengthening was primarily driven by the increased dislocation density, while the contributions of grain refinement were minor. In addition, extra strengthening, which originates from contributions of planar defects (stacking faults/deformation twins), became apparent for greater hot-rolling reductions. The results obtained in this work help in reconsidering the existing strengthening strategy for the alloys, and thus, a novel feasible manufacturing route using conventional hot deformation processing, such as forging, rolling, swaging, and drawing, is realized. STATEMENT OF SIGNIFICANCE: The results obtained in this work suggested a novel microstructural design concept/feasible manufacturing route of high-strength Co-Cr-Mo alloys using conventional hot deformation processing. The present strategy focuses on the strengthening due to the introduction of a high density of lattice defects rather than grain refinement using dynamic recrystallization (DRX). The hot-rolled samples obtained by our process exhibited exceptional strength, which is comparable to the highest strength reported for biomedical Co-Cr-Mo alloys. It was also found that the acceptable ductility can be obtained even in such highly distorted Co-Cr-Mo alloys. We described the strengthening mechanisms in detail; this will be helpful for further investigations or industrial realization of the proposed strategy.

Synthesis of an Iron-Oxidizing System during Growth of Thiobacillus ferrooxidans on Sulfur-Basal Salts Medium.

It was found that the de novo synthesis of not only sulfur:ferric ion oxidoreductase (ferric ion-reducing system) but also iron oxidase was absolutely required when Thiobacillus ferrooxidans AP19-3 was grown on sulfur-salts medium. The results strongly suggest that iron oxidase is involved in sulfur oxidation. This bacterium could not grow on sulfur-salts medium under anaerobic conditions with Fe as a terminal electron acceptor, suggesting that energy conservation by electron transfer between elemental sulfur and Fe is not available for this bacterium.