Gerd Holzhüter

Hydrogen uptake by carbon nanofibers catalyzed by palladium

Abstract Carbon nanofibers of herringbone conformation were obtained by chemical vapor deposition on Pd/La2O3 catalyst, from ethylene—hydrogen mixture. After the removal of La2O3, samples with various Pd/C ratios were obtained by oxidation in air. The hydrogen sorption capacities measured gravimetrically at 10 MPa in pure hydrogen, for six different batches of samples, show a good correlation with the Pd/C ratio reveling a catalytic effect of Pd, which supplies atomic H. A possible charge transfer might lead to the increasing of the H uptake with the increasing Pd/C ratio. A saturation value of 1.5% H (mass) per carbon is reached at rather high Pd/C mole ratio (∼1) for nanofibers with 425– 455 m 2 g −1 BET surface area.

Nanostructuring of Biomaterials—A Pathway to Bone Grafting Substitute

Background:The bone substitute NanoBone® consists of nanocrystalline hydroxyapatite embedded in a highly porous matrix of silica gel. It promotes the healing of bone defects and is degraded by osteoclasts during bone remodeling. The present study investigates the interactions of NanoBone® with bone tissue.Methods:Granules of NanoBone® were implanted in defects of critical size in the mandible of minipigs. Samples were taken after 5 and 10 weeks and demineralized. The composition of the implanted granules was analyzed by means of transmission and scanning electron microscopy and EDX. Enzymeand immunohistochemistry was used to investigate organic components of NanoBone® granules that arised after implantation in the host.Results:EDX demonstrated that 5 weeks after implantation the silica gel was degraded and replaced by an organic matrix. Ultrastructurally, the matrix appeared amorphous with only single collagen fibrillae.PAS-staining indicated the presence of carbohydrates. Immunohistochemically, the bone proteins osteopontin, osteocalcin and BMP-2 were found as constituents of the new matrix. Alkalic phosphatase activity was located in osteoblasts and newly formed bone on NanoBone® and focally in particles. Osteoclasts with ruffled borders, sealing zones, and acid phosphatase activity were situated in resorption lacunae at granule surfaces not covered by new bone.Conclusions:In vivo, the silica gel of NanoBone® is replaced by bone matrix glycoproteins with known functions in attraction, adhesion, and differentation of bone cells as osteoblasts and osteoclasts. We assume that the deposition of these molecules supports the early phase of NanoBone® degradation by osteoclasts and promotes the production of new bone tissue.

Early matrix change of a nanostructured bone grafting substitute in the rat.

A nanocrystalline bone substitute embedded in a highly porous silica gel matrix (NanoBone) has previously been shown to bridge bone defects by an organic matrix. As the initial host response on the bone graft substitute might be a determinant for subsequent bone formation, our present purpose was to characterize the early tissue reaction on this biomaterial. After implantation of 80 mg of NanoBone into the adipose neck tissue of a total of 35 rats, grafts were harvested for subsequent analysis at days 3, 6, 9, 12, and 21. The biomaterial was found encapsulated by granulation tissue which partly penetrated the implant at day 3 and completely pervaded the graft at day 12 on implantation. Histology revealed tartrate-resistant acid phosphatase (TRAP)-positive giant cells covering the biomaterial. ED1 (CD68) immunopositivity of these cells further indicated their osteoclast-like phenotype. Scanning electron microscopy revealed organic tissue components within the periphery of the graft already at day 9, whereas the central hematoma region still presented the silica-surface of the biomaterial. Energy dispersive X-ray spectroscopy further demonstrated that the silica gel was degraded faster in the peripheral granulation tissue than in the central hematoma and was replaced by organic host components by day 12. In conclusion, the silica gel matrix is rapidly replaced by carbohydrate macromolecules. This might represent a key step in the process of graft degradation on its way toward induction of bone formation. The unique composition and structure of this nanoscaled biomaterial seem to support its degradation by host osteoclast-like giant cells.

Biodegradation of titanium implants after long-time insertion used for the treatment of fractured upper and lower jaws through osteosynthesis: element analysis by electron microscopy and EDX or EELS.

Twelve patients underwent an osteosynthesis with titanium to treat upper and lower jaw fractures. Six to 12 months later, the miniplates were removed. Tissue samples were analyzed by light and electron microscopy for detection of a metallosis. The analysis showed new bone formation like callus tissue around the miniplates. In some cases small, rounded deposits and accumulation of colloid-like particles located next to bigger titanium artifacts were detected in the cytoplasm of histiocytes and in the matrix of connective tissue. The titanium was identified by elemental analysis using EDX in SEM as well as by EELS and electron diffraction in TEM. Both kinds of particles contain titanium, but they seem to be different in composition and derivation. The bigger particles seem to consist of metallic titanium and sourced by working on the metallic implants during the implantation itself. On the other hand, the colloidal-like, small, rounded particles in tissue macrophages and outside the cells in the matrix of connective tissue are presumably of other origin; for example, they could be derived from biodegradation and chemical conversion of the metallic implants. The titanium miniplates were examined before and after implantation by using ESCA technique and revealed metallic titanium and different compositions with other elements. The amount of titanium load of the tissue was very low in most cases and presumably not of biomedical relevance.Twelve patients underwent an osteosynthesis with titanium to treat upper and lower jaw fractures. Six to 12 months later, the miniplates were removed. Tissue samples were analyzed by light and electron microscopy for detection of a metallosis. The analysis showed new bone formation like callus tissue around the miniplates. In some cases small, rounded deposits and accumulation of colloid-like particles located next to bigger titanium artifacts were detected in the cytoplasm of histiocytes and in the matrix of connective tissue. The titanium was identified by elemental analysis using EDX in SEM as well as by EELS and electron diffraction in TEM. Both kinds of particles contain titanium, but they seem to be different in composition and derivation. The bigger particles seem to consist of metallic titanium and sourced by working on the metallic implants during the implantation itself. On the other hand, the colloidal-like, small, rounded particles in tissue macrophages and outside the cells in the matrix of connective tissue are presumably of other origin; for example, they could be derived from biodegradation and chemical conversion of the metallic implants. The titanium miniplates were examined before and after implantation by using ESCA technique and revealed metallic titanium and different compositions with other elements. The amount of titanium load of the tissue was very low in most cases and presumably not of biomedical relevance.

Evaluation of injectable silica-embedded nanohydroxyapatite bone substitute in a rat tibia defect model

In clinical practice, vertebral compression fractures occur after trauma and osteoporosis. Kyphoplasty is a minimally invasive procedure using bone filler material for the treatment of such fractures. A full synthetic injectable bone substitute (SIBS) was manufactured by means of spray drying. The aim of this study was to characterize the SIBS and to analyze the remodelling process during degradation of the biomaterial and new bone formation after implantation. SIBS is an aqueous suspension of donut-like microparticles. These microparticles consist of nanocrystallites of synthetic hydroxyapatite embedded in amorphous silica gel. After implantation of SIBS in a proximal tibial diaphyseal defect in 52 rats, grafts were harvested for subsequent analysis on different days. Newly formed bone originating from endosteum was observed on day 6. Hematomas in the medullary space and cortical wounds disappeared on day 12. The wound region was completely replaced by a composite of newly formed cancellous bone, extracellular matrix, and SIBS. At day 63 the cortical defect was fully healed by bone, while newly formed bone in the medullary space almost disappeared and was replaced with bone marrow. In conclusion, SIBS demonstrated a unique structure with osteoinductive and bioresorbable properties, which induced fast bone regeneration. Therefore, a clinical application of SIBS for kyphoplasty is promising.

Lateral augmentation of the mandible in minipigs with a synthetic nanostructured hydroxyapatite block

The purpose of this study was to evaluate biomaterial degradation and new bone formation after implantation of a nanostructured hydroxyapatite (HA) grafting block. Furthermore, physical characteristics of the biomaterial were measured. The biomaterial consists of nanostructured HA embedded in a porous matrix of silica (SiO(2) ) gel. The blocks with two different contents of silica (group A: 24 wt % and group B: 39 wt %) were fixed with titanium screws at the lateral aspect of the mandible of minipigs (n = 5). The specific surface areas of both blocks were measured using Brunauer-Emmett-Teller (BET) equation and mercury intrusion. In all animals, the wound healing was uneventful. After 5 weeks, the biomaterial percentage was 51.5% ± 12.1% for group A and 33.2% ± 5.9% for group B (p = 0.017). New bone formation accounted to 7.6% ± 6.0% for group A and 15.3% ± 8.3% for group B (p = 0.126) after 5 weeks. After 10 weeks, further resorption of the biomaterial led to percentages of 30.6% ± 10.0% for group A and 12.1% ± 6.7% for group B (p = 0.000). After 10 weeks, new bone formations were measured to be 34.1% ± 10.8% in group A and 39.9% ± 13.5% in group B (p = 0.383). The rate of degradation of the biomaterial is controlled by the composition of the material. A higher content of silica gel matrix leads to faster degradation of the biomaterial. The formation of new bone failed to show a significant difference between both groups.

Hydrophobically modified water-soluble polymers and polyelectrolytes as micellar promotors in the Rh(I) catalyzed hydrogenation of an amino acid precursor in water

Hydrophobically modified water-soluble polymers (HMWSPs) and hydrophobically modified polyelectrolytes (HMPEs,“polysoaps”) of various compositions and different molecular weights have been prepared and used in the micellar promoted enantioselective hydrogenation of methyl (Z)-α-acetamidocinnamate with a chiral Rh(I) catalyst in aqueous medium. The efficiency of the micellar polymers in the catalytic reaction is greatly determined by their solubility (dispersibility) behavior and solubilization capacity for catalyst and substrate. The latters depends on the compactness of the polyamphiphiles as revealed by transmission electron microscopy (TEM) and light microscopy images. The highest catalytic performance was obtained with polymer-surfactant complexes prepared by micellar copolymerization with neutral or anionic tensides, regardless of the electrical charge of the hydrophobically modified polymer. In most cases a synergism between rate and enantioselectivity has been observed.

Polymerized ionic amphiphiles: Synthesis and effects in the enantioselective hydrogenation of an amino acid precursor

A number of polymerizable ionic amphiphiles has been polymerized in water at various concentrations above their critical micelle concentration (cmc). Oligomeric and polymeric species are formed upon photochemical initiation. Depending on the position of the double bond within the monomer and polymerization conditions (temperature, concentration, use of radical starters) polymeric assemblies were obtained of different size and hence water-solubility. After separation of low-molecular components by dialysis water-soluble polymers have been characterized by their molecular weights (membrane and vapor pressure osmometry) and transmission electron microscopy (TEM). They showed different efficiency as promotors in the rhodium complex-catalyzed asymmetric hydrogenation of (Z)-methyl α-acetamidocinnamate in water and have been compared with their monomeric counterparts. As was demonstrated in one case, associates formed by intramicellar polymerization display an enhanced catalytic efficiency whereas clustered assemblies are inactive.

Detection of gold particles in the neck skin after lightning stroke with evaporation of an ornamental chain

A German couple was struck by lightning. Both patients survived this event. Whereas the husband was unconscious for only a few minutes, his wife fell into coma for 24 h. The lightning stroke entered the body of the woman behind the left ear and left it at the left shoe. The stroke caused a partial evaporation of a gold ornamental chain on the neck, resulting in a tattoo of the neck skin. A biopsy of the skin 6 months after the event showed the accumulation of gold particles of different size in the dermis down to the subcutaneous fatty tissue. In semithin sections, histiocytes, multinucleated foreign giant cells, and fibroblasts were visible with uptaken metallic particles. In transmission electron microscopy, gold globules of up to 30 µm in diameter were visible outside the cells in the collageneous matrix of the connective tissue besides smaller metallic particles up to 5 nm inside lysosomes and residual bodies of phagocytic cells. Four different kinds of gold particles could be differentiated: globules, granular irregular particles, tubules, and tanglelike tracks. In scanning electron microscopy, gold particles were demonstrated by backscatter detection in the connective tissue of subcutis, where the EDX elemental analysis showed strong signals of aurum (Au), copper (Cu), and argentum (Ag). The detected metals were quantified by AAS as 70% gold, 21% silver, and 9% copper, which demonstrates the composition of gold alloy of the neck chain of the patient. Tanglelike tracks and elongated gold deposits represent crystals of gold salts, as detected by electron diffraction and polarization microscopy. Attempts to remove the gold particles from the skin to remove the tattoo should not be undertaken because the gold is deep and widespread.

Comparison of Bone Substitutes in a Tibia Defect Model in Wistar-Rats

Various bone graft substitutes were used in clinical practise in the treatment of bone defects after trauma or osteoporosis. Many synthetic biomaterials were developed in recent years primarily based on hydroxyapatite (HA). NanoBone® is a nanocrystalline hydroxyapatite (HA) embedded in a porous matrix of silica (SiO2). The ratio of HA:SiO2 varied between 76:24 (wt%; NanoBone®) and 61:39 (wt%; Nanobone® S). The two bone substitutes NB and NB S and a natural bovine bone substitute Bio-Oss® (BO) were evaluated by means of implantation in the tibia of the rat. The aim of this study was to analyze the remodelling process and to measure new bone formation and degradation after implantation of these biomaterials. A tibia defect model was used for all investigations with testing periods of 12, 21 and 84 days. (n=5 for each time point). The results showed, that all bone grafts were well accepted by the host tissue without inflammatory reactions. In comparison to the biomaterial BO, NanoBone® and NanoBone® S were quickly degraded, whereas autologous proteins were incorporated into nanopores. New bone formation was statistically higher in NanoBone® S compared to Bio-Oss® in defect area after 84 days implantation. The presence of osteoclasts in tissue sections were demonstrated by TRAP- and ED1-immunohistology.