Claus Moseke

Determination of the Bone Mineral Crystallite Size and Lattice Strain from Diffraction Line Broadening

Diffraction line broadening observed for the biological apatite is ascribed to small crystallite dimensions and lattice imperfections. However, it is rather difficult to separate the individual contribution of each factor. Therefore in numerous works a total inverse width of a diffraction peak is only used as a size/strain parameter. Several authors determine the bioapatite crystallite size ignoring the lattice strain. As is shown in the present paper, this problem can be resolved for oriented specimens. The crystallite size and lattice strain were calculated by two independent methods: Fourier analysis and approximation with threefold convolution of X-ray lines. The approach proposed can be useful in the investigations into structural aspects of the bone apatite and its synthetic analogues as the crystal size is related to surface defects and the lattice strain to lattice imperfections.

Strontium modified biocements with zero order release kinetics.

Strontium-substituted beta-TCP with the general formula Ca((3-x))Sr(x)(PO(4))(2) (0<x<1) was synthesized by calcination of powder mixtures with the appropriate stoichiometry and reacted with acidic monocalcium phosphate monohydrate (MCPM) to form a cementitious matrix of secondary calcium phosphates. The use of Sr-substituted beta-TCP as a reactant not only induced strontium substitution in the setting products but surprisingly it also favoured the formation of monetite (CaHPO(4)) as setting product, whereas Sr-free cements set to brushite (CaHPO(4).2H(2)O). Release experiments under dynamic conditions for up to 15 days revealed the release of Sr(2+) ions at dose ranges of 12-30 ppm with zero order release kinetic. Cement cytocompatibility was investigated by culturing human osteoblast cell line hFOB1.19 on cement surfaces whereas Sr-containing cements were as good as Sr-free cements in providing a template for cell growth and function.

The effect of Cu(II)-loaded brushite scaffolds on growth and activity of osteoblastic cells

Bone substitute materials such as calcium phosphate cements (CPC) are frequently used as growth factor carriers for the stimulation of osteoblast-formation around an implant. However, biological modification based on delicate protein factors like extracellular matrix proteins or growth factors is subject to a number of shortcomings like the need for storage below room temperature and cost of production. The aim of this study was to investigate ionic modification as an alternative bioinorganic route for implant modification. Although it is known that Cu(II) plays a role in angiogenesis and bone formation, not all involved processes are well understood yet. In this study the in vitro effect of Cu(II) on growth and activity of osteoblastic cells seeded on brushite (CaHPO(4) · 2 H(2) O) scaffolds as well as on glass discs was investigated. The results show that Cu(II) enhances cell activity and proliferation of osteoblastic cells on CPC and furthermore affects the expression of several bone specific proteins such as bone sialo protein or osteocalcin. Therefore, the modification of CPC with Cu(II) may offer a promising alternative to protein based modification to stimulate cellular activity for an improved bone healing.

Hard implant coatings with antimicrobial properties

Infection of orthopaedic implants often leads to inflammation immediately after surgery and increases patient morbidity due to repetitive operations. Silver ions have been shown to combine good biocompatibility with a low risk of inducing bacterial resistance. In this study a physical vapour deposition system using both arc deposition and magnetron sputtering has been utilized to produce silver ion doped TiN coatings on Ti substrates. This biphasic system combines the advantages of silver induced bactericidity with the good mechanical properties of TiN. Crystallographic analysis by X-ray diffraction showed that silver was deposited as well in its elementary form as it was incorporated into the crystal lattice of TiN, which resulted in increasing hardness of the TiN-coatings. Elution experiments revealed a continuous release of Ag ions in phosphate buffered saline. The coatings showed significant inhibitory effects on the growth of Staphylococcus epidermidis and Staphylococcus aureus and practically no cell-toxicity in cytocompatibility tests.

Cu2 +, Co2 + and Cr3 + doping of a calcium phosphate cement influences materials properties and response of human mesenchymal stromal cells

The application of biologically active metal ions to stimulate cellular reactions is a promising strategy to accelerate bone defect healing. Brushite-forming calcium phosphate cements were modified with low doses of Cu2+, Co2+ and Cr3+. The modified cements released the metal ions in vitro in concentrations which were shown to be non-toxic for cells. The release kinetics correlated with the solubility of the respective metal phosphates: 17-45 wt.-% of Co2+ and Cu2+, but <1 wt.-% of Cr3+ were released within 28days. Moreover, metal ion doping led to alterations in the exchange of calcium and phosphate ions with cell culture medium. In case of cements modified with 50mmol Cr3+/mol β-tricalcium phosphate (β-TCP), XRD and SEM analyses revealed a significant amount of monetite and a changed morphology of the cement matrix. Cell culture experiments with human mesenchymal stromal cells indicated that the observed cell response is not only influenced by the released metal ions but also by changed cement properties. A positive effect of modifications with 50mmol Cr3+ or 10mmol Cu2+ per mol β-TCP on cell behaviour was observed in indirect and direct culture. Modification with Co2+ resulted in a clear suppression of cell proliferation and osteogenic differentiation. In conclusion, metal ion doping of the cement influences cellular activities in addition to the effect of released metal ions by changing properties of the ceramic matrix.

Real-time measurement of protein adsorption on electrophoretically deposited hydroxyapatite coatings and magnetron sputtered metallic films using the surface acoustic wave technique.

Surface acoustic wave (SAW) biosensors are highly sensitive for mass binding and are therefore used to detect protein-protein and protein-antibody interactions. Whilst the standard surface of the chips is a thin gold film, measurements on implant- or bone-like surfaces could significantly enhance the range of possible applications for this technique. The aim of this study was to establish methods to coat biosensor chips with Ti, TiN, and silver-doped TiN using physical vapor deposition as well as with hydroxyapatite by electrophoresis. To demonstrate that protein adsorption can be detected on these surfaces, binding experiments with fibronectin and fibronectin-specific antibodies have been performed with the coatings, which successfully proved the applicability of PVD and EPD for SAW biosensor functionalization.

Chemical characterization of hydroxyapatite obtained by wet chemistry in the presence of V, Co, and Cu ions

A model system for the precipitation of hydroxyapatite (HA) from saturated solutions at basic pH was utilized to investigate the effects of V, Co, and Cu ions on crystallography and stoichiometry of the produced apatites. X-ray diffraction (XRD) was applied to analyze phase composition and crystallinity of powders obtained with different metal ion concentrations and annealed at different sintering temperatures. This procedure used the temperature-dependent phase transitions and decompositions of calcium phosphates to analyze the particular influences of the metal ions on apatite mineralization. Comparative XRD measurements showed that all metal ion species reduced crystallinity and crystallite size of the produced apatites. Furthermore the transformation of amorphous calcium phosphate (ACP) to HA was partially inhibited, as was deduced from the formation of α-tricalcium phosphate (α-TCP) peaks in XRD patterns of the heated powders as well as from the reduced intensity of the OH stretch vibration in FTIR spectra. The thermally induced formation of β-TCP indicated a significantly reduced Ca/P ratio as compared to stoichiometric HA. This effect was more pronounced with rising metal ion content. In addition, the appearance of metal oxides in the XRD patterns of samples heated to higher temperatures indicated the incorporation of metal ions in the precipitated apatites. Peak shifts showed that both the apatitic as well as the β-TCP phase apparently had incorporated metal ions.

Electrochemically assisted deposition of strontium modified magnesium phosphate on titanium surfaces.

Electrochemically assisted deposition was utilized to produce ceramic coatings on the basis of magnesium ammonium phosphate (struvite) on corundum-blasted titanium surfaces. By the addition of defined concentrations of strontium nitrate to the coating electrolyte Sr(2+) ions were successfully incorporated into the struvite matrix. By variation of deposition parameters it was possible to fabricate coatings with different kinetics of Sr(2+) into physiological media, whereas the release of therapeutically relevant strontium doses could be sustained over several weeks. Morphological and crystallographic examinations of the immersed coatings revealed that the degradation of struvite and the release of Sr(2+) ions were accompanied by a transformation of the coating to a calcium phosphate based phase similar to low-crystalline hydroxyapatite. These findings showed that strontium doped struvite coatings may provide a promising degradable coating system for the local application of strontium or other biologically active metal ions in the implant-bone interface.

Physical and chemical characterization of Ag-doped Ti coatings produced by magnetron sputtering of modular targets

Silver-doped Ti films were produced using a single magnetron sputtering source equipped with a titanium target containing implemented silver modules under variation of bias voltage and substrate temperature. The Ti(Ag) films were characterized regarding their morphology, contact angle, phase composition, silver content and distribution as well as the elution of Ag(+) ions into cell media. SEM and AFM pictures showed that substrate heating during film deposition supported the formation of even and dense surface layers with small roughness values, an effect that could even be enforced, when a substrate bias voltage was applied instead. The deposition of both Ti and Ag was confirmed by X-ray diffraction. ICP-MS and EDX showed a clear correlation between the applied sputtering parameters and the silver content of the coatings. Surface-sensitive XPS measurements revealed that higher substrate temperatures led to an accumulation of Ag in the near-surface region, while the application of a bias voltage had the opposite effect. Additional elution measurements using ICP-MS showed that the release kinetics depended on the amount of silver located at the film surface and hence could be tailored by variation of the sputter parameters.

Organic/Inorganic Hybrid Nanoceramics Based on Sol-Gel Chemistry

The synthesis of nanostructured organic/inorganic hybrid materials by the sol-gel process plays a key role in the development of advanced materials with suitable properties for, for example, the application in photocatalysis, fuel cells, corrosion protective coatings, and biomedical engineering and biomaterials science. The process is based on the controlled hydrolysis and polycondensation of silica and metal alkoxides, which leads to the formation of nanoscaled particles and their arrangement to a nanoporous network after gelation. Depending on aging and drying conditions, either dense monolithic ceramic composites or (nano)porous materials can be fabricated. This chapter describes recent approaches of bottom-up material fabrication by the sol-gel process including bulk materials like bioactive glasses or dental filling composites and functional coatings in biomedical applications as well as the use of sol-gel-derived materials to encapsulate drugs and even living prokaryotic and eukaryotic cells.