Influence of implant base material on secondary bone healing: an in silico study.

The implant material at the fracture site influences fracture healing not only from biological perspective but also from mechanical perspective. Biodegradable implants such as magnesium (Mg) based alloys have shown faster secondary bone healing properties as compared to bioinert implants such as titanium (Ti). The general reasoning behind this is the benefit of Mg from biocompatibility perspectives. We studied the effect of Ti and Mg as base materials for implants from mechanical perspectives, where we focused on the displacements at the fracture site of the tibia and their influence on the stimulus for bone healing. We found out that in comparison to Ti, Mg implants have minimal stress shielding problem, only which led to better mechanical stimulus at the fracture site.

In vivo urinary compatibility of Mg-Sr-Ag alloy in swine model.

A biodegradable metallic ureteral stent with suitable mechanical properties and antibacterial activity remains a challenge. Here we reveal the scientific significance of a biodegradable Mg-Sr-Ag alloy with a favorable combination of balanced mechanical properties, adjustable indwelling time in urinary tract and evident antibacterial activity via in vivo experiments in a swine model. Attributed to the rheo-solidification process, equiaxial microstructure and significantly refined grains (average grain size: 27.1 µm) were achieved. Mg17Sr2 and Mg4Ag were found as the primary precipitates in the matrix, due to which the alloy obtained ca. 111% increase in ultimate tensile strength in comparison to pure magnesium. Both the in vitro and in vivo results demonstrated the satisfactory biocompatibility of the alloy. Histological evaluation and bioindicators analysis suggested that there was no tissue damage, inflammation and lesions in the urinary system caused by the degradation process. The stent also improved the post-operative bladder functions viewed from the urodynamic results. Our findings highlight the potential of this alloy as antibacterial biodegradable urinary implant material.

Crack Propagation in As-Extruded and Heat-Treated Mg-Dy-Nd-Zn-Zr Alloy Explained by the Effect of LPSO Structures and Their Micro- and Nanohardness.

The investigation of the crack propagation in as-extruded and heat-treated Mg-Dy-Nd-Zn-Zr alloy with a focus on the interaction of long-period stacking-ordered (LPSO) structures is the aim of this study. Solution heat treatment on a hot extruded Mg-Dy-Nd-Zn-Zr (RESOLOY®) was done to change the initial fine-grained microstructure, consisting of grain boundary blocky LPSO and lamellar LPSO structures within the matrix, into coarser grains of less lamellar and blocky LPSO phases. C-ring compression tests in Ringer solution were used to cause a fracture. Crack initiation and propagation is influenced by twin boundaries and LPSO lamellae. The blocky LPSO phases also clearly hinder crack growth, by increasing the energy to pass either through the phase or along its interface. The microstructural features were characterized by micro- and nanohardness as well as the amount and location of LPSO phases in dependence on the heat treatment condition. By applying nanoindentation, blocky LPSO phases show a higher hardness than the grains with or without lamellar LPSO phases and their hardness decreases with heat treatment time. On the other hand, the matrix increases in hardness by solid solution strengthening. The microstructure consisting of a good balance of grain size, matrix and blocky LPSO phases and twins shows the highest fracture energy.

The Video Microscopy-Linked Electrochemical Cell: An Innovative Method to Improve Electrochemical Investigations of Biodegradable Metals.

An innovative, miniature video-optical-electrochemical cell was developed and tested that allows for the conducting of electrochemical corrosion measurements and simultaneous microscopic observations over a small, well-defined surface area of corroding or degrading samples. The setup consisted of a miniature electrochemical cell that was clamped onto the metal sample and fixed under a video microscope before being filled with electrolyte. The miniature cell was comprised of afferent/efferent electrolyte ducts as well as a connection to the Mini Cell System (MCS) for electrochemical measurements. Consequently, all measured and induced currents and voltages referred to the same small area corroding completely within the field of view of the microscope, thus allowing for real-time observation and linking of surface phenomena such as hydrogen evolution and oxide deposition to electrochemical data. The experimental setup was tested on commercial purity (cp) and extra-high purity (XHP) magnesium (Mg) samples using open circuit potential and cyclic voltammetry methods under static and flowing conditions. The corrosion potential was shifted more anodically for cp Mg in comparison to XHP Mg under dynamic conditions. The corrosion current assessed from the cyclic voltametric curves were higher for the cp Mg in comparison to XHP Mg. However, there were no differences between static and flow conditions in the case of XHP Mg in contrast to cp Mg, where the current density was two times higher at dynamic conditions. The measurements and observations with this new method pave the way for a more detailed understanding of magnesium corrosion mechanisms, thus improving predictive power of electrochemical corrosion measurements on newly developed magnesium or other biodegradable alloys applied for medical devices. Different electrochemical tests can be run under various conditions, while being easy to set up and reproduce as well as being minimally destructive to the sample.

Influence of the amount of intermetallics on the degradation of Mg-Nd alloys under physiological conditions.

The influence of amount of intermetallics on the degradation of as-extruded Mg-Nd alloys with different contents of Nd was investigated via immersion testing in DMEM+10% FBS under cell culture conditions and subsequent microstructural characterizations. It is found that the presence of intermetallic particles Mg41Nd5 affects the corrosion of Mg-Nd alloys in two conflicting ways. One is their negative role that their existence enhances the micro-galvanic corrosion. Another is their positive role. Their existence favours the formation of a continuous and compact corrosion layer. At the early stage of immersion, their negative role predominated. The degradation rate of Mg-Nd alloys monotonously increases with increasing the amount of intermetallics. Mg-5Nd alloy with maximum amount of intermetallics suffered from the most severe corrosion. With the immersion proceeding (≥7 days), then the positive role of these intermetallic particles Mg41Nd5 could not be neglected. Owing to the interaction between their positive and negative roles, at the later stage of immersion the corrosion rate of Mg-Nd alloys first increases with increasing the content of Nd, then reaches to the maximum at 2 wt. % Nd. With a further increase of Nd content, a decrease in corrosion rate occurs. The main corrosion products on the surfaces of Mg-Nd alloys include carbonates, calcium-phosphate, neodymium oxide and/or neodymium hydroxide. They are amorphous at the early stage of immersion. With the immersion proceeding, they are transformed to crystalline. The existence of undegradable Mg41Nd5 particles in the corrosion layer can enhance the crystallization of such amorphous corrosion products.

In vivo assessment of biodegradable magnesium alloy ureteral stents in a pig model.

Today, ureteral stent technology is making progress towards the reduction of complications and patient discomfort. Therefore, magnesium alloys have become excellent candidate materials for manufacturing ureteral stents due to their biodegradability and antibacterial activity. Built on our previous work on biodegradable magnesium alloys, this article reports a semisolid rheo-formed magnesium implant that displays degradability and biocompatibility in vivo, and feasibility as ureteral stents in a pig model. Refined non-dendritic microstructure was observed in the rheo-formed alloy, whose grain size and shape factor were ca. 25.2 µm and ca. 1.56 respectively. Neither post-interventional inflammation nor pathological changes were observed in the urinary system during the implantation period of 14 weeks, and the degradation profile (14 weeks) meets the common requirement for the indwelling time of ureteral stents (8 to 16 weeks). Furthermore, histopathological observation and urinalysis results confirmed that the alloy had significantly higher antibacterial activity than the medical-grade stainless steel control. To our knowledge, this is the first in vivo study of biodegradable magnesium alloy as urinary implants in large animal models. Our results demonstrate that magnesium alloys may be a reasonable option for manufacturing biodegradable ureteral stents.

Phase Formation during Solidification of Mg-Nd-Zn Alloys: An In Situ Synchrotron Radiation Diffraction Study.

Mg-4Nd base alloys with Zn additions of 3, 5 and 8 wt % were investigated with in situ synchrotron radiation diffraction during solidification. This method enabled the investigation of phase formation and transformation in the alloys. The diffraction results were supported with TEM observations on the as-solidified samples. The results show the effect of increased Zn addition on stabilizing the Mg3RE phase (RE-rare earth). The experimental results agree only partially with the theoretical calculations indicating the need to improve the existing thermodynamic database on the alloy system.

Microhardness and In Vitro Corrosion of Heat-Treated Mg-Y-Ag Biodegradable Alloy.

Magnesium alloys are promising candidates for biodegradable medical implants which reduce the necessity of second surgery to remove the implants. Yttrium in solid solution is an attractive alloying element because it improves mechanical properties and exhibits suitable corrosion properties. Silver was shown to have an antibacterial effect and can also enhance the mechanical properties of magnesium alloys. Measurements of microhardness and electrical resistivity were used to study the response of Mg-4Y and Mg-4Y-1Ag alloys to isochronal or isothermal heat treatments. Hardening response and electrical resistivity annealing curves in these alloys were compared in order to investigate the effect of silver addition. Procedures for solid solution annealing and artificial aging of the Mg-4Y-1Ag alloy were developed. The corrosion rate of the as-cast and heat-treated Mg-4Y-1Ag alloy was measured by the mass loss method. It was found out that solid solution heat treatment, as well artificial aging to peak hardness, lead to substantial improvement in the corrosion properties of the Mg-4Y-1Ag alloy.

Influence of the Microstructure and Silver Content on Degradation, Cytocompatibility, and Antibacterial Properties of Magnesium-Silver Alloys In Vitro.

Implantation is a frequent procedure in orthopedic surgery, particularly in the aging population. However, it possesses the risk of infection and biofilm formation at the surgical site. This can cause unnecessary suffering to patients and burden on the healthcare system. Pure Mg, as a promising metal for biodegradable orthopedic implants, exhibits some antibacterial effects due to the alkaline pH produced during degradation. However, this antibacterial effect may not be sufficient in a dynamic environment, for example, the human body. The aim of this study was to increase the antibacterial properties under harsh and dynamic conditions by alloying silver metal with pure Mg as much as possible. Meanwhile, the Mg-Ag alloys should not show obvious cytotoxicity to human primary osteoblasts. Therefore, we studied the influence of the microstructure and the silver content on the degradation behavior, cytocompatibility, and antibacterial properties of Mg-Ag alloys in vitro. The results indicated that a higher silver content can increase the degradation rate of Mg-Ag alloys. However, the degradation rate could be reduced by eliminating the precipitates in the Mg-Ag alloys via T4 treatment. By controlling the microstructure and increasing the silver content, Mg-Ag alloys obtained good antibacterial properties in harsh and dynamic conditions but had almost equivalent cytocompatibility to human primary osteoblasts as pure Mg.

Influence of Dy in solid solution on the degradation behavior of binary Mg-Dy alloys in cell culture medium.

Rare earth element Dy is one of the promising alloying elements for magnesium alloy as biodegradable implants. To understand the effect of Dy in solid solution on the degradation of Mg-Dy alloys in simulated physiological conditions, the present work studied the microstructure and degradation behavior of Mg-Dy alloys in cell culture medium. It is found the corrosion resistance enhances with the increase of Dy content in solid solution in Mg. This can be attributed to the formation of a relatively more corrosion resistant Dy-enriched film which decreases the anodic dissolution of Mg.

Microstructure and degradation performance of biodegradable Mg-Si-Sr implant alloys.

In this work the microstructure and degradation behavior of several as-cast alloy compositions belonging to the Mg rich corner of the Mg-Si-Sr system are presented and related. The intermetallic phases are identified and analyzed describing the microstructure evolution during solidification. It is intended in this work to obtain insight in the behavior of the ternary alloys in in vitro tests and to analyze the degradation behavior of the alloys under physiologically relevant conditions. The as-cast specimens have been exposed to immersion tests, both mass loss (ML) and potentiodynamic polarization (PDP). The degradation rate (DR) have been assessed and correlated to microstructure features, impurity levels and alloy composition. The initial reactions resulted to be more severe while the degradation stabilizes with time. A higher DR is related with a high content of the Mg17Sr2 phase and with the presence of coarse particles of the intermetallics Mg2Si, MgSiSr and MgSi2Sr. Specimens with a higher DR typically have higher levels of impurities and alloy contents.

Intramedullary Mg2Ag nails augment callus formation during fracture healing in mice.

UNLABELLED: Intramedullary stabilization is frequently used to treat long bone fractures. Implants usually remain unless complications arise. Since implant removal can become technically very challenging with the potential to cause further tissue damage, biodegradable materials are emerging as alternative options. Magnesium (Mg)-based biodegradable implants have a controllable degradation rate and good tissue compatibility, which makes them attractive for musculoskeletal research. Here we report for the first time the implantation of intramedullary nails made of an Mg alloy containing 2% silver (Mg2Ag) into intact and fractured femora of mice. Prior in vitro analyses revealed an inhibitory effect of Mg2Ag degradation products on osteoclast differentiation and function with no impair of osteoblast function. In vivo, Mg2Ag implants degraded under non-fracture and fracture conditions within 210days and 133days, respectively. During fracture repair, osteoblast function and subsequent bone formation were enhanced, while osteoclast activity and bone resorption were decreased, leading to an augmented callus formation. We observed a widening of the femoral shaft under steady state and regenerating conditions, which was at least in part due to an uncoupled bone remodeling. However, Mg2Ag implants did not cause any systemic adverse effects. These data suggest that Mg2Ag implants might be promising for intramedullary fixation of long bone fractures, a novel concept that has to be further investigated in future studies. STATEMENT OF SIGNIFICANCE: Biodegradable implants are promising alternatives to standard steel or titanium implants to avoid implant removal after fracture healing. We therefore developed an intramedullary nail using a novel biodegradable magnesium-silver-alloy (Mg2Ag) and investigated the in vitro and in vivo effects of the implants on bone remodeling under steady state and fracture healing conditions in mice. Our results demonstrate that intramedullary Mg2Ag nails degrade in vivo over time without causing adverse effects. Importantly, radiographs, µCT and bone histomorphometry revealed a significant increase in callus size due to an augmented bone formation rate and a reduced bone resorption in fractures supported by Mg2Ag nails, thereby improving bone healing. Thus, intramedullary Mg2Ag nails are promising biomaterials for fracture healing to circumvent implant removal.

An in vivo study on the metabolism and osteogenic activity of bioabsorbable Mg-1Sr alloy.

Previous studies indicated that local delivery of strontium effectively increased bone quality and formation around osseointegrating implants. Therefore, implant materials with long-lasting and controllable strontium release are avidly pursued. The central objective of the present study was to investigate the in vivo biocompatibility, metabolism and osteogenic activity of the bioabsorbable Mg-1Sr (wt.%, nominal composition) alloy for bone regeneration. The general corrosion rate of the alloy implant as a femoral fracture fixation device was 0.55±0.03mm·y(-1) (mean value±standard deviation) in New Zealand White rabbits which meet the bone implantation requirements and can be adjusted by material processing methods. All rabbits survived and the histological evaluation showed no abnormal physiology or diseases 16 weeks post-implantation. The degradation process of the alloy did not significantly alter 16 primary indexes of hematology, cardiac damage, inflammation, hepatic functions and metabolic process. Significant increases in peri-implant bone volume and direct bone-to-implant contact (48.3%±15.3% and 15.9%±5.6%, respectively) as well as the expressions of four osteogenesis related genes (runt-related transcription factor 2, alkaline phosphatase, osteocalcin, and collagen, type I, alpha 1) were observed after 16 weeks implantation for the Mg-1Sr group when compared to the pure Mg group. The sound osteogenic properties of the Mg-1Sr alloy by long-lasting and controllable Sr release suggesting a very attractive clinical potential. STATEMENT OF SIGNIFICANCE: Sr (strontium) has exhibited pronounced effects to reduce the bone fracture risk in osteoporotic patients. Nonetheless, long-lasting local Sr release is hardly achieved by traditional methods like surface treatment. Therefore, a more efficient Sr local delivery platform is in high clinical demand. The stable and adjustable degradation process of Mg alloy makes it an ideal Sr delivery platform. We combine the well-known osteogenic properties of strontium with magnesium to manufacture bioabsorbable Mg-1Sr alloy with stable Sr release based on our previous studies. The in vitro and in vivo results both showed the alloy's suitable degradation rate and biocompatibility, and the sound osteogenic properties and stimulation effect on bone formation suggest its very attractive clinical potential.

Histological Comparison of New Biodegradable Magnesium-Based Implants for Maxillofacial Applications.

BACKGROUND: A variety of materials have been used for bone augmentation, distraction osteotomy, and in post-cancer patients following tumor removal. However, a temporary metal implant that would resorb after successful treatment is a new concept. Magnesium was suggested as a suitable material for these purposes because it is biocompatible, has better mechanical properties than titanium, and stimulates new bone formation. This study evaluates histological appearance of magnesium-based implants and the surrounding bone. MATERIALS AND METHODS: Three magnesium-based biomaterials were tested in a rabbit bone defect model: magnesium-hydroxyapatite (Mg-HA), W4 (96 % magnesium, 4 % yttrium), and pure magnesium (pure Mg). Animals were sacrificed after 6 and 12 weeks and the samples were analyzed histologically and histomorphometrically. RESULTS: Mg-HA had the highest mean amount of tartrate-resistant acid phosphatase (TRAP) positive cells at the implantation site of all groups. It had shown the fastest degradation rate already at 6 weeks but the least amount of new bone formation. New bone was seen forming in direct contact with pure Mg and W4. The mean gas volume was highest in W4 compared to pure Mg and Mg-HA but this difference was not statistically significant. W4 had the lowest mean number of TRAP-positive cells of all materials. CONCLUSION: Pure Mg and W4 were shown to be the most promising materials in this study in respect to the bone response to the implant material. They could be used for screws and plates in bone augmentation procedures.

Mechanical properties and corrosion behavior of Mg-Gd-Ca-Zr alloys for medical applications.

Magnesium alloys are promising candidates for biomedical applications. In this work, influences of composition and heat treatment on the microstructure, the mechanical properties and the corrosion behavior of Mg-Gd-Ca-Zr alloys as potential biomedical implant candidates were investigated. Mg5Gd phase was observed at the grain boundaries of Mg-10Gd-xCa-0.5Zr (x=0, 0.3, 1.2wt%) alloys. Increase in the Ca content led to the formation of additional Mg2Ca phase. The Ca additions increased both the compressive and the tensile yield strengths, but reduced the ductility and the corrosion resistance in cell culture medium. After solution heat treatment, the Mg5Gd particles dissolved in the Mg matrix. The compressive strength decreased, while the corrosion resistance improved in the solution treated alloys. After ageing at 200°C, metastable ß' phase formed on prismatic planes and a new type of basal precipitates have been observed, which improved the compressive and tensile ultimate strength, but decreased the ductility.

Element distribution in the corrosion layer and cytotoxicity of alloy Mg-10Dy during in vitro biodegradation.

The present work investigates the corrosion behaviour, the element distribution in the corrosion layer and the cytocompatibility of alloy Mg-10Dy. The corrosion experiments were performed in a cell culture medium (CCM) under cell culture conditions close to the in vivo environment. The element distribution on the surface as well as in cross-sections of the corrosion layer was investigated using scanning electron microscopy, energy-dispersive X-ray analysis, X-ray photoelectron spectroscopy and X-ray diffraction. The cytocompatibility of alloy Mg-10Dy with primary human osteoblasts was evaluated by MTT, cell adhesion and live/dead staining tests. The results show that the corrosion layer was enriched in Dy, while the P and Ca content gradually decreased from the surface to the bottom of the corrosion layer. In addition, large amounts of MgCO3·3H2O formed in the corrosion layer after 28 days immersion. Both extracts and the Dy-enriched corrosion layer of alloy Mg-10Dy showed no cytotoxicity to primary human osteoblasts.

Fast escape of hydrogen from gas cavities around corroding magnesium implants.

Magnesium materials are of increasing interest in the development of biodegradable implants as they exhibit properties that make them promising candidates. However, the formation of gas cavities after implantation of magnesium alloys has been widely reported in the literature. The composition of the gas and the concentration of its components in these cavities are not known as only a few studies using non-specific techniques were done about 60 years ago. Currently many researchers assume that these cavities contain primarily hydrogen because it is a product of magnesium corrosion in aqueous media. In order to clearly answer this question we implanted rare earth-containing magnesium alloy disks in mice and determined the concentration of hydrogen gas for up to 10 days using an amperometric hydrogen sensor and mass spectrometric measurements. We were able to directly monitor the hydrogen concentration over a period of 10 days and show that the gas cavities contained only a low concentration of hydrogen gas, even shortly after formation of the cavities. This means that hydrogen must be exchanged very quickly after implantation. To confirm these results hydrogen gas was directly injected subcutaneously. Most of the hydrogen gas was found to exchange within 1h after injection. Overall, our results disprove the common misbelief that these cavities mainly contain hydrogen and show how quickly this gas is exchanged with the surrounding tissue.

Influence of ageing treatment on microstructure, mechanical and bio-corrosion properties of Mg-Dy alloys.

Mg-Dy alloys have shown to be promising for medical applications. In order to investigate the influence of ageing treatment on their mechanical and corrosion properties, three Mg-xDy alloys (x=10, 15, 20 wt%) were prepared. Their microstructure, mechanical and corrosion behavior were investigated. The results indicate that ageing at 250 °C has little influence on the mechanical and corrosion properties. In contrast, ageing at 200 °C significantly increases the yield strength, and reduces the ductility. After ageing at 200 °C, the corrosion rate of Mg-20Dy alloy increases largely in 0.9 wt% NaCl solution, but remains unchanged in cell culture medium.

Ion release from magnesium materials in physiological solutions under different oxygen tensions.

Although magnesium as degradable biomaterial already showed clinical proof of concepts, the design of new alloys requires predictive in vitro methods, which are still lacking. Incubation under cell culture conditions to obtain "physiological" corrosion may be a solution. The aim of this study was to analyse the influence of different solutions, addition of proteins and of oxygen availability on the corrosion of different magnesium materials (pure Mg, WE43, and E11) with different surface finishing. Oxygen content in solution, pH, osmolality and ion release were determined. Corrosion led to a reduction of oxygen in solution. The influence of oxygen on pH was enhanced by proteins, while osmolality was not influenced. Magnesium ion release was solution-dependent and enhanced in the initial phase by proteins with delayed release of alloying elements. The main corrosion product formed was magnesium carbonate. Therefore, cell culture conditions are proposed as first step toward physiological corrosion.