Microstructure, mechanical properties, biocompatibility, and in vitro corrosion and degradation behavior of a new Zn-5Ge alloy for biodegradable implant materials.

Zinc (Zn)-based alloys are considered a new class of biodegradable implant materials due to their superior chemical stability and processability compared to biodegradable magnesium (Mg) alloys. In this study, we report a new biodegradable Zn-5Ge alloy with highly desirable mechanical, corrosion, and biological properties. Microstructural characterization revealed the effective grain-refining effect of germanium (Ge) on the Zn alloy. Tensile test results indicated that the hot-rolled Zn-5Ge alloy showed an ultimate tensile strength of 237.0 MPa, a yield strength of 175.1 MPa, and an elongation of 21.6%; while as-cast pure Zn showed an ultimate tensile strength of 33.6 MPa, a yield strength of 29.3 MPa, and an elongation of 1.2%. The corrosion rates measured by potentiodynamic polarization tests in Hank's solution in ascending order are: as-cast Zn-5Ge (0.1272 mm/y) < as-cast pure Zn (0.1567 mm/y) < hot-rolled Zn-5Ge (0.2255 mm/y) < hot-rolled pure Zn (0.3057 mm/y). Immersion tests revealed that the degradation rate of as-cast Zn-5Ge is 0.042 mm/y, less than half of that of hot-rolled pure Zn and ∼62% of that of as-cast pure Zn. Moreover, the Zn-5Ge alloy showed excellent in vitro hemocompatibility and the addition of 5% Ge effectively enhanced the hemocompatibility of pure Zn. CCK-8 assay using murine preosteoblast MC3T3-E1 cells indicated that the diluted extracts at a concentration <12.5% of both the as-cast Zn-5Ge alloy and pure Zn showed grade 0 cytotoxicity; the diluted extracts at the concentrations of 50% and 25% of Zn-5Ge alloy showed a significantly higher cell viability than those of pure Zn. STATEMENT OF SIGNIFICANCE: Zinc (Zn)-based alloys are currently considered a new class of biodegradable implant materials due to their excellent processability. Here, we report a novel Zn-5Ge alloy with highly desirable mechanical, corrosion and biological properties. The tensile test results indicated that the hot-rolled Zn-5Ge alloy showed an ultimate tensile strength of 237.0 MPa, a yield strength of 175.1 MPa and an elongation of 21.6%; while as-cast pure Zn showed an ultimate tensile strength of 33.6 MPa, a yield strength of 29.3 MPa and an elongation of 1.2%. The corrosion rate measured by potentiodynamic polarization tests in Hank's solution in the ascending order is: as-cast Zn-5Ge (0.1272 mm/y) < as-cast pure Zn (0.1567 mm/y) < hot-rolled Zn-5Ge (0.2255 mm/y) < hot-rolled pure Zn (0.3057 mm/y). Immersion tests revealed that the degradation rate of the as-cast Zn-5Ge is 0.042 mm/y, less than half of that of the hot-rolled pure Zn, ∼62% of that of as-cast pure Zn. Moreover, the Zn-5Ge alloy showed excellent in vitro biocompatibility.

Effects of citric acid and the siderophore desferrioxamine B (DFO-B) on the mobility of germanium and rare earth elements in soil and uptake in Phalaris arundinacea.

Effects of citric acid and desferrioxamine B (DFO-B) on the availability of Ge and selected rare earth elements (REEs) (La, Nd, Gd, Er) to Phalaris arundinacea were investigated. A soil dissolution experiment was conducted to elucidate the effect of citric acid and DFO-B at different concentrations (1 and 10 mmol L-1 citric acid) on the release of Ge and REEs from soil. In a greenhouse, plants of P. arundinacea were cultivated on soil and on sand cultures to investigate the effects of citric acid and DFO-B on the uptake of Ge and REEs by the plants. Addition of 10 mmol L-1 citric acid significantly enhanced desorption of Ge and REEs from soil and uptake into soil-grown plants. Applying DFO-B enhanced the dissolution and the uptake of REEs, while no effect on Ge was observed. In sand cultures, the presence of citric acid and DFO-B significantly decreased the uptake of Ge and REEs, indicating a discrimination of the formed complexes during uptake. This study clearly indicates that citric acid and the microbial siderophore DFO-B may enhance phytoextraction of Ge and REEs due to the formation of soluble complexes that increase the migration of elements in the rhizosphere.

Intrinsically germanium-69-labeled iron oxide nanoparticles: synthesis and in-vivo dual-modality PET/MR imaging.

Intrinsically germanium-69-labeled super-paramagnetic iron oxide nanoparticles are synthesized via a newly developed, fast and highly specific chelator-free approach. The biodistribution pattern and the feasibility of (69) Ge-SPION@PEG for in vivo dual-modality positron emission tomography/magnetic resonance (PET/MR) imaging and lymph-node mapping are investigated, which represents the first example of the successful utilization of a (69) Ge-based agent for PET/MR imaging.

Metabolism of tellurium, antimony and germanium simultaneously administered to rats.

Recently, tellurium (Te), antimony (Sb) and germanium (Ge) have been used as an alloy in phase-change optical magnetic disks, such as digital versatile disk-random access memory (DVD-RAM) and DVD-recordable disk (DVD-RW). Although these metalloids, the so-called "exotic" elements, are known to be non-essential and harmful, little is known about their toxic effects and metabolism. Metalloid compounds, tellurite, antimonite and germanium dioxide, were simultaneously administered to rats. Their distributions metabolites were determined and identified by speciation. Te and Sb accumulated in red blood cells (RBCs): Te accumulated in RBCs in the dimethylated form, while Sb accumulated in the inorganic/non-methylated form. In addition, trimethyltelluronium (TMTe) was the urinary metabolite of Te, whereas Sb in urine was not methylated but oxidized. Ge was also not methylated in rats. These results suggest that each metalloid is metabolized via a unique pathway.

Dermal absorption of inorganic germanium in rats.

So-called germanium 'health' products including dietary supplements, cosmetics, accessories, and warm bath service containing germanium compounds and metalloid are popular in Japan. Subchronic and chronic oral exposure of germanium dioxide (GeO(2)), popular chemical form of inorganic germanium causes severe germanium toxicosis including death and kidney dysfunction in humans and experimental animals. Intestinal absorption of neutralized GeO(2) or germanate is almost complete in humans and animals. However, it is not known whether germanium is cutaneously absorbed. We tested dermal absorption of neutralized GeO(2) or germanate using male F344/N rats. Three groups of rats were treated with a 3-h topical application of hydrophilic ointment containing graded level of neutralized GeO(2) (pH 7.4): 0, 0.21 and 0.42 mg GeO(2)/g. Germanium concentration in blood and tissues sampled from rats after topical application of inorganic germanium was measured by inductively coupled plasma-mass spectrometry. Animals topically applied 0.42 mg GeO(2)/g ointment had significantly higher germanium concentrations in plasma, liver, and kidney than those of rats that received no topical germanium. The results indicate that skin is permeable to inorganic germanium ion or germanate and recurrent exposure of germanium compounds may pose a potential health hazard.

Biological fabrication of photoluminescent nanocomb structures by metabolic incorporation of germanium into the biosilica of the diatom Nitzschia frustulum.

Diatoms are single-celled algae that make microscale silica shells or "frustules" with intricate nanoscale features such as two-dimensional pore arrays. In this study, the metabolic insertion of low levels of germanium into the frustule biosilica of the pennate diatom Nitzschia frustulum by a two-stage cultivation process induced the formation of frustules which strongly resembled double-sided nanocomb structures. The final product from the two-stage cultivation process contained 0.41 wt % Ge in biosilica and consisted of an equal mixture of parent frustule valves possessing a normal two-dimensional array of 200 nm pores and daughter valves possessing the nanocomb structure. The nanocomb structures had overall length of 8 mum, rib width of 200 nm, rib length of 500 nm, and slit width of 100 nm. Each slit of the nanocomb was most likely formed by a directed morphology change of a row of 200 nm pores to a single open slit following Ge incorporation into the developing frustule during the final cell division. The frustules possessed blue photoluminescence at peak wavelengths between 450 and 480 nm, which was attributed to contributions from nanostructured biosilica in both the parent valves and in the nanocomb daughter valves. This is the first reported study of using a cell culture system to biologically fabricate a photoluminescent nanocomb structure.

Sensitivity and daily quality control of a mobile PET/CT scanner operating in 3-dimensional mode.

UNLABELLED: This study investigated the stability of the sensitivity of a mobile PET/CT scanner and tested a phantom experiment to improve on the daily quality control recommendations of the manufacturer. Unlike in-house scanners, mobile PET/CT devices are subjected to a harsher, continuously changing environment that can alter their performance. The parameter of sensitivity was investigated because it reflects directly on standardized uptake value, a key factor in cancer evaluation. METHODS: A (68)Ge phantom of known activity concentration was scanned 6 times a month for 11 consecutive months using a mobile PET/CT scanner that operates in 3-dimensional mode only. The scans were acquired as 2 contiguous bed positions, with raw data obtained and reconstructed using parameters identical to those used for oncology patients, including CT-extracted attenuation coefficients and decay, scatter, geometry, and randoms corrections. After visual inspection of all reconstructed images, identical regions of interest were drawn on each image to obtain the activity concentration of individual slices. The original activity concentration was then decay-corrected to the scanning day, and the percentage sensitivity of the slice was calculated and graphed. The daily average sensitivity of the scanner, over 11 consecutive months, was also obtained and used to evaluate the stability of sensitivity. RESULTS: Our particular scanner showed a daily average sensitivity ranging from -8.6% to 6.5% except for one instance, when the sensitivity dropped by an unacceptable degree, 34.8%. CONCLUSION: Our 11-mo follow-up of a mobile PET/CT scanner demonstrated that its sensitivity remained within acceptable clinical limits except for one instance, when the scanner had to be serviced before patients could be imaged. To enhance our confidence in the uniformity of sensitivity across slices, we added a phantom scan to the daily quality control recommendations of the manufacturer.

Determination of germanium in urine and its usefulness for biomonitoring of inhalation exposure to inorganic germanium in the occupational setting.

The present study aimed to assess whether urinary germanium concentration can be used as a biomarker of inhalation exposure to airborne dust from metallic germanium (Ge) or GeO2 in the occupational setting. A novel hydride generation-based method coupled with fow-injection graphite furnace atomic absorption spectrometry (HG/FI-GFAAS) was developed for the determination of urinary germanium. It was found that urinary germanium concentration could be reliably determined by a standard additions method after thorough digestion of the urine and careful pH adjustment of the digest. The limit of detection (LOD) in urine for the HG/FI-GFAAS method was 0.25 microg Ge L(-1). In Belgian control male subjects, the urinary germanium concentration was below this LOD. In 75 workers currently exposed to inorganic germanium compounds, respirable and inhalable concentrations of germanium in the aerosols were measured on Monday and Friday at the job sites using personal air samplers. Spot-urine samples were collected on the same days before and after the work shift. The germanium concentrations of respirable dust correlated very well with those of inhalable dust and represented 20% of the inhalable fraction. Workers exposed to metallic Ge dust were on average ten times less exposed to germanium than those whose exposure involved GeO2 (3.4 versus 33.8 microg Ge m(-3)). This difference was reflected in the urinary germanium concentrations (3.4 versus 23.4 microg Ge g(-1) creatinine). Regression analysis showed that the concentration of germanium in the inhalable fraction explained 42% of the post-shift urinary germanium concentration either on Monday or on Friday, whereas in a subgroup of 52 workers mainly exposed to metallic germanium dust 57% (r = 0.76) of the Monday post-shift urinary germanium was explained. Urinary elimination kinetics were studied in seven workers exposed to airborne dust of either metallic Ge or GeO2. The urinary elimination rate of germanium was characterised by half-times ranging from 8.2 to 18.1 h (on average 12 h 46 min). The present study did not allow discrimination between the germanium species to which the workers were exposed, but it showed fast urinary elimination kinetics for inhalation exposure to dust of metallic Ge and GeO2. It pointed out that urine samples taken at the end of the work shift can be used for biological monitoring of inorganic germanium exposure in the occupational setting.

Determination of germanium in human specimens: comparative study of atomic absorption spectrometry and microwave-induced plasma mass spectrometry.

The determination methods of germanium (Ge) in biological specimens such as blood plasma, erythrocytes, urine, hair, nail, and other organs were established using graphite furnace atomic absorption spectrometry (GFAAS) and microwave-induced plasma mass spectrometry (MIP-MS). The detection limits of Ge standard solution were 3 ng/mL with GFAAS and 0.05 ng/mL with MIP-MS. The detection limits in organ samples depended on the type of samples and sampling amounts: 3-30 ng/g by GFAAS and 0.05-0.5 ng/g by MIP-MS. The sensitivity of GFAAS was lower than that of MIP-MS; however, it was adequate for determining Ge concentrations in specimens from patients who had ingested Ge. Samples were digested by a simple wet-ashing procedure using nitric acid and perchloric acid. To avoid the interfering effects of coexisting elements and perchloric acid residue, an extraction method using organic solvent was tried. When using MIP-MS, extraction was not necessary; however, both dilution and addition of an internal standard were needed. Special attention was required for iron-rich samples because a molecular ion of 56Fe16O was observed at nm/z72 where 2Ge was monitored. The results of Ge concentrations in human samples obtained by these methods agreed well. Interfering effects of perchloric acid, which was used for digestion and which remained in samples, were observed in both methods. Hair and nail samples from people who had ingested Ge were useful for monitoring Ge in the body. Hair samples were useful for determining past exposure to Ge when the distribution patterns from the scalp to the end of the strand were analyzed. In control subjects, Ge concentrations in the listed specimens and organs were lower than 0.1 microg/g or mL, and these low levels of Ge were able to be determined by MIP-MS in combination with the extraction method.

Kinetics of germanium dioxide in rats.

The kinetics of germanium dioxide (GeO2) in single dose and repeated exposures were investigated in male Wistar rats. In the single dose GeO2 (100 mg/kg BW, p.o.) exposure study, values of several kinetic parameters were shown as follows, a maximum concentration in serum of 15.5+/-0.7 microg/ml (mean +/- S.E.M.), an absorption half-life of 0.7+/-0.1 h (mean +/- S.E.M.), an elimination half-life of 2.3+/-0.5 h (mean +/- S.E.M.), a distribution of the central compartment Vp (3.1+/-0.3 1, mean +/- S.E.M.), and the apparent volume of distribution of the tissue compartment Vt (8.5+/-2.9 1, mean +/- S.E.M.). In the repeated exposure study, 730+/-92 mg GeO2 in 1 1 double-distilled H2O ( = 100 mg/kg/day) was given daily to rats for 4 weeks (p.o.). After sacrificing the rats, the analysis of tissue distribution showed that GeO2 was accumulated in some important organs or tissues in the body, especially the peripheral nerves and kidney. These results indicate that GeO2 could be absorbed rapidly but had a longer elimination half-life in rats. In addition, GeO2 was accumulated especially in the nerves and kidney following long-term exposure.

Pharmacokinetics of germanium after po beta-carboxyethylgermanium sesquioxide in 24 Chinese volunteers.

AIM: To compare the pharmacokinetics after po different doses of beta-carboxyethylgermanium sesquioxide (Ge-132). METHODS: An atomic absorption spectrophotometric system was used to measure germanium concentrations in plasma and urine samples after po Ge-132 1 (low dose, LD), 2.5 (medium dose, MD), and 4 (high dose, HD) g.m-2 in 24 healthy volunteers (one dose per 8 subjects). RESULTS: T1/2 alpha (LD, 1.2 +/- 0.7 h; MD, 1.1 +/- 0.6 h; HD, 1.2 +/- 0.5 h), T1/2 beta (LD, 5.2 +/- 1.2 h; MD, 5.8 +/- 2.5 h; HD, 5.5 +/- 1.4 h) and Cl/F (LD, 33 +/- 12 L.h-1; MD, 35 +/- 10 L.h-1; HD, 33 +/- 11 L.h-1) were not dose-related. Tmax was between 0.75 h and 2 h. Cmax (LD, 5.3 +/- 2.2 mg.L-1; MD, 13 +/- 5 mg.L-1; HD 18 +/- 8 mg.L-1, HD) and AUC (LD, 31 +/- 13 mg.h.L-1; MD, 60 +/- 16 mg.h.L-1; HD, 79 +/- 42 mg.h.L-1) were positive correlation to the dose of Ge-132. Urine-eliminated germanium within 24 h accounted for 11 +/- 3% of LD, 9 +/- 3% of MD, and 6 +/- 5% of HD (calculated from Ge/F) and showed a negative correlation to the dose. CONCLUSION: 1) Intracorporal process of Ge after po Ge-132 coincided with the first-order absorption and elimination with two-compartment kinetic model; 2) The amount of germanium eliminated in urine was below 11%.

Dissolution/reprecipitation and protein adsorption studies of calcium phosphate coatings by FT-IR/ATR techniques.

The surfaces of bioactive Ca-P ceramics immediately change when exposed to proteinaceous solutions. The dissolution behavior and protein interactions of these bioactive materials at the bone/implant interface need to be investigated to understand their material-cellular interactions fully. In this study, FT-IR/ATR techniques were used to study the in situ phosphate release kinetics of Ca-P coatings. The net loss of phosphate molecules from coatings was slower in saline solutions compared with alpha-MEM solutions. Coatings exposed to alpha-MEM solutions containing fibronectin released phosphate molecules slower than coatings exposed to alpha-MEM solutions containing albumin. Conformational changes in fibronectin and albumin adsorbed onto Ca-P and uncoated germanium surfaces were also investigated using FT-IR/ATR spectroscopy. Analysis of changes in the amide I bands indicated that there was a greater loss of beta-sheet structure in adsorbed fibronectin on Ca-P coatings when compared with bare germanium surfaces. Although albumin did change its structure upon adsorption on both Ca-P and germanium, unlike fibronectin, adsorbed albumin structure was similar on Ca-P coatings and germanium. Furthermore, with time the conformation of adsorbed fibronectin and albumin appeared to be very stable on Ca-P coatings, whereas albumin adsorbed to germanium exhibited an increase in ratio of alpha-helix to beta-turn.

[Absolute and relative bioavailability of germanium in the rabbit]. / Biodisponibilités absolue et relative du germanium chez le lapin.

The debated consumption of germanium suggested the authors to compare biopharmaceutical parameters of germanium oxide and germanium sesquioxide. A first evaluation, in rabbit, has been based on Germanium blood levels determined by atomic absorption spectrometry, after cross administration of both products by the I.V. and oral routes. When given orally, the apparent oxide bioavailability is very low (about 10%) but better than that of the sesquioxide. That difference could result from differences of disposition parameters of both products, which have to be studied late.

Chronic tubulointerstitial changes induced by germanium dioxide in comparison with carboxyethylgermanium sesquioxide.

Chronic nephrotoxicity was investigated in rats orally administered germanium dioxide (GeO2) and carboxyethylgermanium sesquioxide (Ge-132) for 24 weeks. Increased BUN and serum phosphate as well as decreased creatinine clearance, weight loss, anemia and liver dysfunction were apparent at week 24 only in the GeO2 treated group. Vacuolar degeneration and granular depositions were observed by light microscope in the degenerated renal distal tubules in the rats of this group, with the semiquantitative scores of tubular degeneration being 95 +/- 9% in the GeO2 group, 3 +/- 1% in the Ge-132 group and 1 +/- 1% in the control group, respectively. Electron microscopy revealed electron-dense inclusions in the swollen mitochondrial matrix of the distal tubular epithelium in the GeO2 group. Although systemic toxicities were reduced after GeO2 was discontinued at week 24, renal tubulointerstitial fibrosis became prominent even at week 40 (16 weeks after discontinuation). A Ge.K alpha X-ray spectrum was clearly demonstrated in the mitochondrial matrix of the distal tubular epithelium in the GeO2 group with the help of electron probe X-ray microanalysis. On the other hand, neither toxic effects nor renal histological abnormalities were manifested in either the Ge-132 or the control group. The renal tissue content of germanium was high at weeks 24 and 40 in the GeO2 group. From these results, it is concluded that GeO2 causes characteristic nephropathy while Ge-132 does not. In addition, it appears that residual GeO2 remains for a considerably long time even after the cessation of GeO2 intake.

Subacute nephrotoxicity of germanium dioxide in the experimental animal.

Germanium (Ge; atomic number 32, atomic weight 72.6) belongs to IVb group of the Periodic Table and is found as a trace metal in soil, rocks, plants, and animals. It is widely used in industry because of its semiconductive nature. Some biological activities have been shown in Ge derivatives. Recently, patients with persistent renal damage after chronic ingestion of germanium dioxide (GeO2)-containing compounds have been reported in Japan. This study aimed to investigate subacute nephrotoxicity of GeO2 in Lewis male rats. The rats were treated orally with GeO2 for 13 weeks (GeO2 group) and were compared with those treated with GeO2 for only the first 4 weeks (GeO2-4-week group) and with untreated controls. Renal dysfunction was demonstrated by the increased serum creatinine, BUN, and serum phosphate and decreased creatinine clearance. Liver dysfunction was observed as demonstrated by the increased GOT and GPT, and hypoproteinemia by the decreased total protein and albumin in the GeO2 group. However, daily urinary protein excretion or urinalysis did not differ among the groups. Kidney weight and Ge content of tissues were significantly elevated in the GeO2 group. With the light microscope, vacuoles and the depositions of PAS-stained particles, which correspond to electron-microscopic dense granules in the swollen mitochondria, were predominantly observed in distal tubular epithelium in the GeO2 group. Even in the GeO2-4-week group of rats, serum creatinine was increased and the above-mentioned histological abnormalities were observed, but were less intense.

Determination and biokinetics of germanium in mouse tissues by atomic absorption spectrometry with electrothermal atomization.

There are few papers on the analysis of Ge in biomaterials. A highly sensitive method is required for such determination of Ge because its concentration is very low. We developed a sensitive analytical method for determining Ge in biomaterials by using 0.05 mol/l Co (NO3)2 media as a matrix modifier with flameless atomic absorption spectrometry. This method was applied to the determination of Ge in mouse tissues, and the biokinetic parameters of Ge in mouse tissues were evaluated using the data of uptake and retention after a single peroral administration of GeO2 solution. Ge was not detectable in the blood or tissues of normal mice. The uptake of Ge into kidney is much larger than that into other tissues after injection of GeO2 solution. The disappearance of Ge from blood and the tissues is very fast. It could hardly be detected in any of the tissues 24 h after injection. The biological half-life of Ge in brain is 6.3 h, but in other tissues--1.2 h (in blood) to 4.5 h (in pancreas)--it is shorter than in brain. By the fact that Ge is rapidly excreted from all tissues, it can be understood that Ge is not detected in normal mouse tissues and its toxicity is weaker than that of other metals.

Germanium-induced nephropathy: report of two cases and review of the literature.

We report two cases of renal failure following long-term ingestion of germanium dioxide (GeO2) and comment on eight other cases reported in Japan. Ge-induced nephropathy is characterized by insidious onset of renal failure without proteinuria or hematuria after oral intake of Ge-containing compounds for more than several months, and by degeneration of renal tubular cells with minor glomerular abnormality in histology. When patients ceased to ingest Ge compounds, renal function gradually recovered but never returned to the normal range. Serious extrarenal complication can contribute to an unfavorable prognosis.