Assessment of tissue response in vivo: PET-CT imaging of titanium and biodegradable magnesium implants.

To study in vivo the bioactivity of biodegradable magnesium implants and other possible biomaterials, we are proposing a previously unexplored application of PET-CT imaging, using available tracers to follow soft tissue and bone remodelling and immune response in the presence of orthopaedic implants. Female Wistar rats received either implants (Ti6Al7Nb titanium or WE43 magnesium) or corresponding transcortical sham defects into the diaphyseal area of the femurs. Inflammatory response was followed with [18F]FDG and osteogenesis with [18F]NaF, over the period of 1.5 months after surgery. An additional pilot study with [68Ga]NODAGA-RGD tracer specific to αvß3 integrin expression was performed to follow the angiogenesis for one month. [18F]FDG tracer uptake peaked on day 3 before declining in all groups, with Mg and Ti groups exhibiting overall higher uptake compared to sham. This suggests increased cellular activity and tissue response in the presence of Mg during the initial weeks, with Ti showing a subsequent increase in tracer uptake on day 45, indicating a foreign body reaction. [18F]NaF uptake demonstrated the superior osteogenic potential of Mg compared to Ti, with peak uptake on day 7 for all groups. [68Ga]NODAGA-RGD pilot study revealed differences in tracer uptake trends between groups, particularly the prolonged expression of αvß3 integrin in the presence of implants. Based on the observed differences in the uptake trends of radiotracers depending on implant material, we suggest that PET-CT is a suitable modality for long-term in vivo assessment of orthopaedic biomaterial biocompatibility and underlying tissue reactions. STATEMENT OF SIGNIFICANCE: The study explores the novel use of positron emission tomography for the assessment of the influence that biomaterials have on the surrounding tissues. Previous related studies have mostly focused on material-related effects such as implant-associated infections or to follow the osseointegration in prosthetics, but the use of PET to evaluate the materials has not been reported before. The approach tests the feasibility of using repeated PET-CT imaging to follow the tissue response over time, potentially improving the methodology for adopting new biomaterials for clinical use.

Assessment of the Roles of Magnesium and Zinc in Clinical Disorders.

The ability and facility of magnesium (Mg2+) and zinc (Zn2+) to interact with phosphate ions confer them the characteristics of essential trace elements. Trace elements are extremely necessary for the basic nucleic acid chemistry of cells of all known living organisms. More than 300 enzymes require zinc and magnesium ions for their catalytic actions, including all the enzymes involved in the synthesis of ATP. In addition, enzymes such as isomerases, oxidoreductases, lyases, transferases, ligases and hydrolases that use other nucleotides to synthesize DNA and RNA require magnesium and zinc. These nucleotides may trigger oxidative damage or important changes against free radicals. In the same way, nucleotides may play an important role in the pathophysiology of degenerative diseases, including in some clinical disorders, where vascular risk factors, oxidative stress and inflammation work to destabilize the patients` homeostatic equilibrium. Indeed, reduced levels of zinc and magnesium may lead to inadequate amount of antioxidant enzymes, and thus, acts as an important contributing factor for the induction of oxidative stress leading to cellular or tissue dysfunction. Hence, the development of zinc or magnesium enzyme inhibitors could be a novel opportunity for the treatment of some human disorders. Therefore, the objective of the present work was to assess the clinical benefits of zinc and magnesium in human health and their effects in some clinical disorders.

In vitro and in vivo assessment of decellularized platelet-rich fibrin-loaded strontium doped porous magnesium phosphate scaffolds in bone regeneration.

The present work reports the effect of decellularized platelet-rich fibrin (dPRF) loaded strontium (Sr) doped porous magnesium phosphate (MgP) bioceramics on biocompatibility, biodegradability, and bone regeneration. Sustained release of growth factors from dPRF is a major objective here, which conformed to the availability of dPRF on the scaffold surface even after 7 days of in vitro degradation. dPRF-incorporated MgP scaffolds were implanted in the rabbit femoral bone defect and bone rejuvenation was confirmed by radiological examination, histological examination, fluorochrome labeling study, and micro-CT. µ-CT examination of the regained bone samples exhibited that invasion of mature bone in the pores of the MgP2Sr-dPRF sample was higher than the MgP2Sr which indicated better bone maturation capability of this composition. Quantifiable assessment using oxytetracycline labeling showed 73.55 ± 1.12% new osseous tissue regeneration for MgP2Sr-dPRF samples in contrast to 65.47 ± 1.16% for pure MgP2Sr samples, after 3 months of implantation. Histological analysis depicted the presence of abundant osteoblastic and osteoclastic cells in dPRF-loaded Sr-doped MgP samples as compared to other samples. Radiological studies also mimicked similar results in the MgP2Sr-dPRF group with intact periosteal lining and significant bridging callus formation. The present results indicated that dPRF-loaded Sr-doped magnesium phosphate bioceramics have good biocompatibility, bone-forming ability, and suitable biodegradability in bone regeneration.

In vitro and in vivo assessment of the effect of biodegradable magnesium alloys on osteogenesis.

Magnesium (Mg) and some of its alloys are considered promising biodegradable metallic biomaterials for bone implant applications. The osteogenesis effect of Mg alloys is widely reported; however, the underlying mechanisms are still not clear. In this study, pure Mg, Mg-3Zn, and Mg-2Zn-1Mn were prepared, and their degradation behavior, biocompatibility, and osteogenesis effect were systematically assessed both in vitro and in vivo. Primary rat bone marrow-derived mesenchymal stem cells (BMSCs) were used to evaluate the biocompatibility of the prepared Mg alloys, and a rat femur fracture model was used to assess the stimulating effect of these alloys on bone-tissue formation. Mg-2Zn-1Mn showed higher corrosion resistance and more stable degradation behavior than pure Mg and Mg-3Zn. Extracts of the three materials showed significant stimulating effects on osteogenic differentiation of BMSCs along with non-cytotoxicity. Implantation of Mg-2Zn-1Mn wires into the femur of rats demonstrated superior histocompatibility, stable degradation, and notable promotion of osteogenesis without systemic toxicity. Moreover, the results of both in vitro and in vivo assessments demonstrated that bone morphogenetic proteins and fibroblast growth factor receptors are involved in the stimulating effect of Mg alloys. STATEMENT OF SIGNIFICANCE: This work reports the degradation behavior, biocompatibility, and osteogenic effect of pure Mg and Mg-3Zn and Mg-2Zn-1Mn alloys in both in vitro and in vivo conditions. Mg-2Zn-1Mn showed higher corrosion resistance and more stable degradation behavior than pure Mg and Mg-3Zn. The extracts of the three materials showed a significant stimulating effect on osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells (BMSCs) along with non-cytotoxicity. Mg-2Zn-1Mn wires implanted into the femur of rats showed good histocompatibility, stable degradation, and notable promotion of osteogenesis without systemic toxicity. The results of the present study suggest that bone morphogenetic proteins (BMPs) and fibroblast growth factor receptors (FGFRs) are involved in the stimulating effect of Mg alloys on osteogenesis.

In vitro and in vivo degradation assessment and preventive measures of biodegradable Mg alloys for biomedical applications.

Magnesium (Mg) and its alloys have been widely explored as a potential biodegradable implant material. However, the fast degradation of Mg-based alloys under physiological environment has hindered their widespread use for implant applications till date. The present review focuses on in vitro and in vivo degradation of biodegradable Mg alloys, and preventive measures for biomedical applications. Initially, the corrosion assessment approaches to predict the degradation behavior of Mg alloys are discussed along with the measures to control rapid corrosion. Furthermore, this review attempts to explore the correlation between in vitro and in vivo corrosion behavior of different Mg alloys. It was found that the corrosion depends on experimental conditions, materials and the results of different assessment procedures hardly matches with each other. It has been demonstrated the corrosion rate of magnesium can be tailored by alloying elements, surface treatments and heat treatments. Various researches also studied different biocompatible coatings such as dicalcium phosphate dihydrate (DCPD), ß-tricalcium phosphate (ß-TCP), hydroxyapatite (HA), polycaprolactone (PCL), polylactic acid (PLA), and so on, on Mg alloys to suppress rapid degradation and examine their influence on new bone regeneration as well. This review shows the need for a standard method of corrosion assessment to predict the in vivo corrosion rate based on in vitro data, and thus reducing the in vivo experimentation.

Assessment and Imaging of Intracellular Magnesium in SaOS-2 Osteosarcoma Cells and Its Role in Proliferation.

Magnesium is an essential nutrient involved in many important processes in living organisms, including protein synthesis, cellular energy production and storage, cell growth and nucleic acid synthesis. In this study, we analysed the effect of magnesium deficiency on the proliferation of SaOS-2 osteosarcoma cells. When quiescent magnesium-starved cells were induced to proliferate by serum addition, the magnesium content was 2-3 times lower in cells maintained in a medium without magnesium compared with cells growing in the presence of the ion. Magnesium depletion inhibited cell cycle progression and caused the inhibition of cell proliferation, which was associated with mTOR hypophosphorylation at Serine 2448. In order to map the intracellular magnesium distribution, an analytical approach using synchrotron-based X-ray techniques was applied. When cell growth was stimulated, magnesium was mainly localized near the plasma membrane in cells maintained in a medium without magnesium. In non-proliferating cells growing in the presence of the ion, high concentration areas inside the cell were observed. These results support the role of magnesium in the control of cell proliferation, suggesting that mTOR may represent an important target for the antiproliferative effect of magnesium. Selective control of magnesium availability could be a useful strategy for inhibiting osteosarcoma cell growth.

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.

Contributions and competition of Mg2+ and K+ in folding and stabilization of the Twister ribozyme.

Native folded and compact intermediate states of RNA typically involve tertiary structures in the presence of divalent ions such as Mg2+ in a background of monovalent ions. In a recent study, we have shown how the presence of Mg2+ impacts the transition from partially unfolded to folded states through a "push-pull" mechanism where the ion both favors and disfavors the sampling of specific phosphate-phosphate interactions. To further understand the ion atmosphere of RNA in folded and partially folded states results from atomistic umbrella sampling and oscillating chemical potential grand canonical Monte Carlo/molecular dynamics (GCMC/MD) simulations are used to obtain atomic-level details of the distributions of Mg2+ and K+ ions around Twister RNA. Results show the presence of 100 mM Mg2+ to lead to increased charge neutralization over that predicted by counterion condensation theory. Upon going from partially unfolded to folded states, overall charge neutralization increases at all studied ion concentrations that, while associated with an increase in the number of direct ion-phosphate interactions, is fully accounted for by the monovalent K+ ions. Furthermore, K+ preferentially interacts with purine N7 atoms of helical regions in partially unfolded states, thereby potentially stabilizing the helical regions. Thus, both secondary helical structures and formation of tertiary structures leads to increased counterion condensation, thereby stabilizing those structural features of Twister. Notably, it is shown that K+ can act as a surrogate for Mg2+ by participating in specific interactions with nonsequential phosphate pairs that occur in the folded state, explaining the ability of Twister to self-cleave at submillimolar Mg2+ concentrations.

Mechanical performance and biocompatibility assessment of Zn-0.05wt%Mg-(0.5, 1 wt%) Ag alloys.

Zn alloys are emerging as promising degradable biomedical materials due to their tailorable mechanical properties and moderate biodegradable rate, compared with conventional biodegradable metallic materials. Ag, as an effective antibacterial and reinforcing element, was incorporated into Zn-0.05Mg alloys. In the present work, the effects of the Ag addition on mechanical, cytotoxic, hemolytic, pyrogenic, histological behaviors of the animal were investigated. The compressive yielding strength is enhanced from 198 MPa for Zn-0.05Mg alloy up to 224 and 234 MPa for Zn-0.05Mg-0.5Ag and Zn-0.05Mg-1Ag alloys, respectively. When the compressive strain was 65%, the strength of the Zn-0.05Mg-1.0Ag alloy reached 833 MPa, which was much higher than that of 721 MPa for Zn-0.05Mg alloy. The relative growth rate (RGR) for the extracts of Zn-0.05Mg-1Ag alloy with the concentrations of 10, 50, and 100% after 5 days incubation reaches 98.5, 95.2, and 94.2%, which are higher than those in extracts of Zn-0.05Mg-0.5Ag alloy (98.2, 93.9, 92.1%). The hemolysis rate of the Zn-0.05Mg alloys with 0.5 and 1 wt% Ag is 2.46 and 2.28%, respectively. The variations of body weight and temperature, postinjection symptoms, pathological morphologies of the visceral organs demonstrate that the alloys are nontoxic according to the toxicity rating standards. Zn-0.05wt%Mg-(0.5, 1 wt%) Ag alloys are experimentally safe materials and promising for the future application as biodegradable medical devices.

Antidepressant and antinociceptive activities in animal models and in vitro assessment of the anti-thyroid activity of bis(DL-pyroglutamate)magnesium complex.

BACKGROUND: Magnesium is an essential element related with biochemistry of the brain and different types of depression have been associated with its deficiency. METHODS: The structure of a novel magnesium bis(DL-pyroglutamate) (Mg(DL-pGlu)2) was elucidated by X-ray crystallography. Wistar rats were used in the in vivo experiments. The antidepressant-like effect was assessed by the forced swim test (FST) and the antinociceptive activity was evaluated using hot plate test. In both, non-specific effects were evaluated by the open field test. Anti-thyroid activity was examined using Lang's method. Albumin binding behavior was evaluated by 3D fluorescence spectroscopy. RESULTS: For the Mg(DL-pGlu)2 complex (30 mg/kg), the FST test on Wistar rats revealed a decrease of 22% in the immobility time and an increment of 106% in the swimming time. The compound alters neither the locomotor activity nor the body weight after chronic administration. At the same dose, it showed antinociceptive activity, increasing the response latency. It blocks iodination reactions generating a charge transfer complex with iodine hence indicating anti-thyroid activity (Kc = 45366.5 ±â€¯29 M-1). Albumin 3D fluorescence spectroscopy experiments showed intensity increase of peak A and decrease of peak B. CONCLUSIONS: The results showed that the new compound produced a lowering of the immobility time and an increment of the swimming ability of the rats. The compound is able to increase the response latency in 70.0%, to capture iodine (anti-thyroid activity) and to interact with albumin through covalent type of interaction of the free NH groups.

Magnesium-zinc scaffold loaded with tetracycline for tissue engineering application: In vitro cell biology and antibacterial activity assessment.

Recently, porous magnesium and its alloys are receiving great consideration as biocompatible and biodegradable scaffolds for bone tissue engineering application. However, they presented poor antibacterial performance and corrosion resistance which limited their clinical applications. In this study, Mg-Zn (MZ) scaffold containing different concentrations of tetracycline (MZ-xTC, x = 1, 5 and 10%) were fabricated by space holder technique to meet the desirable antibacterial activity and corrosion resistance properties. The MZ-TC contains total porosity of 63-65% with pore sizes in the range of 600-800 µm in order to accommodate bone cells. The MZ scaffold presented higher compressive strength and corrosion resistance compared to pure Mg scaffold. However, tetracycline incorporation has less significant effect on the mechanical and corrosion properties of the scaffolds. Moreover, MZ-xTC scaffolds drug release profiles show an initial immediate release which is followed by more stable release patterns. The bioactivity test reveals that the MZ-xTC scaffolds are capable of developing the formation of HA layers in simulated body fluid (SBF). Next, Staphylococcus aureus and Escherichia coli bacteria were utilized to assess the antimicrobial activity of the MZ-xTC scaffolds. The findings indicate that those scaffolds that incorporate a high level concentration of tetracycline are tougher against bacterial organization than MZ scaffolds. However, the MTT assay demonstrates that the MZ scaffolds containing 1 to 5% tetracycline are more effective to sustain cell viability, whereas MZ-10TC shows some toxicity. The alkaline phosphatase (ALP) activity of the MZ-(1-5)TC was considerably higher than that of MZ-10TC on the 3 and 7 days, implying higher osteoblastic differentiation. All the findings suggest that the MZ-xTC scaffolds containing 1 to 5% tetracycline is a promising candidate for bone tissue healing due to excellent antibacterial activity and biocompatibility.

Dantrolene requires Mg2+ to arrest malignant hyperthermia.

Malignant hyperthermia (MH) is a clinical syndrome of skeletal muscle that presents as a hypermetabolic response to volatile anesthetic gases, where susceptible persons may develop lethally high body temperatures. Genetic predisposition mainly arises from mutations on the skeletal muscle ryanodine receptor (RyR). Dantrolene is administered to alleviate MH symptoms, but its mechanism of action and its influence on the Ca2+ transients elicited by MH triggers are unknown. Here, we show that Ca2+ release in the absence of Mg2+ is unaffected by the presence of dantrolene but that dantrolene becomes increasingly effective as cytoplasmic-free [Mg2+] (free [Mg2+]cyto) passes mM levels. Furthermore, we found in human muscle susceptible to MH that dantrolene was ineffective at reducing halothane-induced repetitive Ca2+ waves in the presence of resting levels of free [Mg2+]cyto (1 mM). However, an increase of free [Mg2+]cyto to 1.5 mM could increase the period between Ca2+ waves. These results reconcile previous contradictory reports in muscle fibers and isolated RyRs, where Mg2+ is present or absent, respectively, and define the mechanism of action of dantrolene is to increase the Mg2+ affinity of the RyR (or "stabilize" the resting state of the channel) and suggest that the accumulation of the metabolite Mg2+ from MgATP hydrolysis is required to make dantrolene administration effective in arresting an MH episode.

Cytotoxicity assessment of adipose-derived mesenchymal stem cells on synthesized biodegradable Mg-Zn-Ca alloys.

Magnesium (Mg)-based alloys have been extensively considered as biodegradable implant materials for orthopedic surgery. Mg and its alloys are metallic biomaterials that can degrade in the body and promote new bone formation. In this study, the corrosion behavior and cytotoxicity of Mg-Zn-Ca alloys are evaluated with adipose-derived mesenchymal stem cells (ASCs). Mg-2Zn and Mg-2Zn-xCa (x=1, 2 and 3wt.%) alloys were designated. Mg alloys were analyzed with scanning electron microscopy and potentiodynamic polarization. To understand the in-vitro biocompatibility and cytotoxicity of Mg-2Zn and Mg-2Zn-xCa alloys, ASCs were cultured for 24 and 72h in contact with 10%, 50% and 100% extraction of all alloys prepared in DMEM. Cell cytotoxicity and viability of ASCs were examined by MTT assay. Alloying elements including Zn and Ca improved the corrosion resistance of alloys were compared with pure Mg. The cytotoxicity results showed that all alloys had no significant adverse effects on cell viability in 24h. After 72h, cell viability and proliferation increased in the cells exposed to pure Mg and Mg-2Zn-1Ca extracts. The release of Mg, Zn and Ca ions in culture media had no toxic impacts on ASCs viability and proliferation. Mg-2Zn-1Ca alloy can be suggested as a good candidate to be used in biomedical applications.

Uranium in well drinking water of Kabul, Afghanistan and its effective, low-cost depuration using Mg-Fe based hydrotalcite-like compounds.

Toxic elements in drinking water have great effects on human health. However, there is very limited information about toxic elements in drinking water in Afghanistan. In this study, levels of 10 elements (chromium, nickel, copper, arsenic, cadmium, antimony, barium, mercury, lead and uranium) in 227 well drinking water samples in Kabul, Afghanistan were examined for the first time. Chromium (in 0.9% of the 227 samples), arsenic (7.0%) and uranium (19.4%) exceeded the values in WHO health-based guidelines for drinking-water quality. Maximum chromium, arsenic and uranium levels in the water samples were 1.3-, 10.4- and 17.2-fold higher than the values in the guidelines, respectively. We next focused on uranium, which is the most seriously polluted element among the 10 elements. Mean ± SD (138.0 ± 1.4) of the 238U/235U isotopic ratio in the water samples was in the range of previously reported ratios for natural source uranium. We then examined the effect of our originally developed magnesium (Mg)-iron (Fe)-based hydrotalcite-like compounds (MF-HT) on adsorption for uranium. All of the uranium-polluted well water samples from Kabul (mean ± SD = 190.4 ± 113.9 µg/L; n = 11) could be remediated up to 1.2 ± 1.7 µg/L by 1% weight of our MF-HT within 60 s at very low cost (<0.001 cents/day/family) in theory. Thus, we demonstrated not only elevated levels of some toxic elements including natural source uranium but also an effective depurative for uranium in well drinking water from Kabul. Since our depurative is effective for remediation of arsenic as shown in our previous studies, its practical use in Kabul may be encouraged.

Degradation of bioabsorbable Mg-based alloys: Assessment of the effects of insoluble corrosion products and joint effects of alloying components on mammalian cells.

This work is focused on the processes occurring at the bioabsorbable metallic biomaterial/cell interfaces that may lead to toxicity. A critical analysis of the results obtained when degradable metal disks (pure Mg and rare earth-containing alloys (ZEK100 alloys)) are in direct contact with cell culture and those obtained with indirect methods such as the use of metal salts and extracts was made. Viability was assessed by Acridine Orange dye, neutral red and clonogenic assays. The effects of concentration of corrosion products and possible joint effects of the binary and ternary combinations of La, Zn and Mg ions, as constituents of ZEK alloys, were evaluated on a mammalian cell culture. In all cases more detrimental effects were found for pure Mg than for the alloys. Experiments with disks showed that gradual alterations in pH and in the amount of corrosion products were better tolerated by cells and resulted in higher viability than abrupt changes. In addition, viability was dependent on the distance from the source of ions. Experiments with extracts showed that the effect of insoluble degradation products was highly detrimental. Indirect tests with Zn ions revealed that harmful effects may be found at concentrations ≥ 150 µM and at ≥ 100 µM in mixtures with Mg. These mixtures lead to more deleterious effects than single ions. Results highlight the need to develop a battery of tests to evaluate the biocompatibility of bioabsorbable biomaterials.

Optimization of a low-cost defined medium for alcoholic fermentation--a case study for potential application in bioethanol production from industrial wastewaters.

In bioethanol production processes, the media composition has an impact on product concentration, yields and the overall process economics. The main purpose of this research was to develop a low-cost mineral-based supplement for successful alcoholic fermentation in an attempt to provide an economically feasible alternative to produce bioethanol from novel sources, for example, sugary industrial wastewaters. Statistical experimental designs were used to select essential nutrients for yeast fermentation, and its optimal concentrations were estimated by Response Surface Methodology. Fermentations were performed on synthetic media inoculated with 2.0 g L(-1) of yeast, and the evolution of biomass, sugar, ethanol, CO2 and glycerol were monitored over time. A mix of salts [10.6 g L(-1) (NH4)2HPO4; 6.4 g L(-1) MgSO4·7H2O and 7.5 mg L(-1) ZnSO4·7H2O] was found to be optimal. It led to the complete fermentation of the sugars in less than 12h with an average ethanol yield of 0.42 g ethanol/g sugar. A general C-balance indicated that no carbonaceous compounds different from biomass, ethanol, CO2 or glycerol were produced in significant amounts in the fermentation process. Similar results were obtained when soft drink wastewaters were tested to evaluate the potential industrial application of this supplement. The ethanol yields were very close to those obtained when yeast extract was used as the supplement, but the optimized mineral-based medium is six times cheaper, which favorably impacts the process economics and makes this supplement more attractive from an industrial viewpoint.

Assessment of cellular reactions to magnesium as implant material in comparison to titanium and to glyconate using the mouse tail model.

PURPOSE: Nowadays, research in magnesium alloys as a biodegradable implant material has increased. The aim of this study was to examine osteoinductive properties and tissue responses to pure magnesium in comparison to conventional permanent (titanium) and to degradable (glyconate) implant materials. METHODS: Magnesium wires (0.4 mm in diameter, 10 mm length) were implanted into tail veins of mice and examined after 2, 4, 8, 16 and 32 weeks. Titanium and glyconate as controls were assessed after 2, 4, 8 and 24 weeks. µ-computed tompgraphy, histology and SEM examinations were performed. RESULTS: Magnesium implants showed increasing structural losses over time with fragmentation after an observation period of 32 weeks. Glyconate was fully degraded and titanium remained almost unaffected after 24 weeks. In contrast to some titanium and glyconate implants, first calcium and phosphate precipitations could be observed around magnesium implants after two weeks. However, ossification could not be observed even after 32 weeks, whereas enchondral ossification was found partially in the sourrounding of glyconate and titanium implants after eight weeks. Nevertheless, magnesium implants showed less inflammatory responses and fibrosing properties than the conventional implant materials. CONCLUSIONS: Although the assumed osteoinductive properties could not be detected, magnesium appears to be a promising degradable implant material because of the low sensitizing and inflammatory potential.

The high heart health value of drinking-water magnesium.

Universal drinking water and beverages containing moderate to high levels of magnesium (10­100 ppm) could potentially prevent 4.5 million heart disease and stroke deaths per year, worldwide. This potential is calculated with 2010 global mortality figures combined with a recent quantification of water-magnesium's inverse association with heart disease and stroke mortality. The modern processed food diet, low in magnesium and spreading globally, makes this well-researched potential of drinking-water magnesium worth serious consideration, especially in areas where insufficient dietary intake of magnesium is prevalent.

Luminal Mg2+, a key factor controlling RYR2-mediated Ca2+ release: cytoplasmic and luminal regulation modeled in a tetrameric channel.

In cardiac muscle, intracellular Ca(2+) and Mg(2+) are potent regulators of calcium release from the sarcoplasmic reticulum (SR). It is well known that the free [Ca(2+)] in the SR ([Ca(2+)](L)) stimulates the Ca(2+) release channels (ryanodine receptor [RYR]2). However, little is known about the action of luminal Mg(2+), which has not been regarded as an important regulator of Ca(2+) release. The effects of luminal Ca(2+) and Mg(2+) on sheep RYR2 were measured in lipid bilayers. Cytoplasmic and luminal Ca(2+) produced a synergistic increase in the opening rate of RYRs. A novel, high affinity inhibition of RYR2 by luminal Mg(2+) was observed, pointing to an important physiological role for luminal Mg(2+) in cardiac muscle. At diastolic [Ca(2+)](C), luminal Mg(2+) inhibition was voltage independent, with K(i) = 45 microM at luminal [Ca(2+)] ([Ca(2+)](L)) = 100 microM. Luminal and cytoplasmic Mg(2+) inhibition was alleviated by increasing [Ca(2+)](L) or [Ca(2+)](C). Ca(2+) and Mg(2+) on opposite sides of the bilayer exhibited competitive effects on RYRs, indicating that they can compete via the pore for common sites. The data were accurately fitted by a model based on a tetrameric RYR structure with four Ca(2+)-sensing mechanisms on each subunit: activating luminal L-site (40-microM affinity for Mg(2+) and Ca(2+)), cytoplasmic A-site (1.2 microM for Ca(2+) and 60 microM for Mg(2+)), inactivating cytoplasmic I(1)-site (approximately 10 mM for Ca(2+) and Mg(2+)), and I(2)-site (1.2 microM for Ca(2+)). Activation of three or more subunits will cause channel opening. Mg(2+) inhibition occurs primarily by Mg(2+) displacing Ca(2+) from the L- and A-sites, and Mg(2+) fails to open the channel. The model predicts that under physiological conditions, SR load-dependent Ca(2+) release (1) is mainly determined by Ca(2+) displacement of Mg(2+) from the L-site as SR loading increases, and (2) depends on the properties of both luminal and cytoplasmic activation mechanisms.

Electric fields due to synaptic currents sharpen excitatory transmission.

The synaptic response waveform, which determines signal integration properties in the brain, depends on the spatiotemporal profile of neurotransmitter in the synaptic cleft. Here, we show that electrophoretic interactions between AMPA receptor-mediated excitatory currents and negatively charged glutamate molecules accelerate the clearance of glutamate from the synaptic cleft, speeding up synaptic responses. This phenomenon is reversed upon depolarization and diminished when intracleft electric fields are weakened through a decrease in the AMPA receptor density. In contrast, the kinetics of receptor-mediated currents evoked by direct application of glutamate are voltage-independent, as are synaptic currents mediated by the electrically neutral neurotransmitter GABA. Voltage-dependent temporal tuning of excitatory synaptic responses may thus contribute to signal integration in neural circuits.