Mineralized dental tissues and dental pulp were characterized using advanced analytical transmission electron microscopy (TEM) methods. Quantitative X-ray energy dispersive spectroscopy was employed to determine the Ca/P and Mg/P concentration ratios. Significantly lower Ca/P concentration ratios were measured in peritubular dentine compared to intertubular dentine, which is accompanied by higher and variable Mg/P concentration ratios. There is strong evidence that magnesium is partially substituting calcium in the hydroxyapatite structure. Electron energy-loss near-edge structures (ELNES) of C-K and O-K from enamel and dentine are noticeably different. We observe a strong influence of beam damage on mineralized dental tissues and dental pulp, causing changes of the composition and consequently also differences in the ELNES. In this article, the importance of TEM sample preparation and specimen damage through electron irradiation is demonstrated.

Teeth are an excellent example where optimally designed nanoarchitectures with precisely constructed components consist of simple compounds. Typically, these simple constituent phases with insignificant properties show mechanical property amplifications when formed into composite architectures. Material properties of functional composites are generally regulated on the nanoscale, which makes their characterization particularly demanding. Using advanced analytical and imaging transmission electron microscopy techniques, we identified innovative microstructural adjustments combined with astonishing compositional adaptations in incisors of coypu. Unique constituents, recognized as an additional amorphous Fe-rich surface layer followed by a transition zone covering pigmented enamel, provide the required structural stability to withstand repeated mechanical load. The chemically diverse Fe-rich surface layer, including ferrihydrite and iron-calcium phosphates, gives the typical orange-brown coloration to the incisors. Within the spaces between elongated hydroxyapatite crystals in the pigmented enamel, only ferrihydrite was found, implying that enamel pigmentation is a very strictly controlled process. Most significantly, an unprecedentedly high amount of Mg was measured in the amorphous flake-like material within the dentinal tubules of the incisors, suggesting the presence of a (Mg,Ca) phosphate phase. This unusually high influx of Mg into the dentin of incisors, but not molars, suggests a substantial functionality of Mg in the initial formation stages and constant growth of incisors. The present results emphasize the strong mutual correlation among the microstructure, chemical composition, and mechanical properties of mineralized dental tissues.

Exposed regions of the arthropod exoskeleton have specialized structure and mineral composition. Their study can provide insights into the evolutionary optimization of the cuticle as a material. We determined the structural and compositional features of claws in the crustacean Ligia pallasii using X-ray micro-computed tomography, scanning electron microscopy (SEM), and analytical scanning transmission electron microscopy (STEM). In addition, we used nanoindentation to determine how these features fine-tune the mechanical properties of the claw cuticle. We found that the inner layer of the claw cuticle—the endocuticle—contains amorphous calcium phosphate, while the outer layer—the exocuticle—is not mineralized and contains elevated amounts of bromine. While the chitin–protein fibers in crustacean exoskeletons generally shift their orientation, they are aligned axially in the claws of L. pallasii. As a consequence, the claw cuticle has larger elastic modulus and hardness in the axial direction. We show that amorphous calcium phosphate mineralization and the brominated cuticle are widespread in isopod crustaceans inhabiting terrestrial habitats. We discuss how the features of the claw cuticle may aid in minimizing the likelihood of fracture. Ultimately, our study points out the features that increase the durability of thin skeletal elements.

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Up to now, chemical synthesis routes only provide restricted opportunities for the formation of structured nano particles. In contrast, living microorganisms generate nano materials of well defined shapes by the precise control of biomineralization. Here we reveal new principles for the generation of functional nano materials through the process of biomineralization. We used the detoxification mechanism of the unicellular alga <i>Scenedesmus obliquus</i> to generate a techno logically interesting zinc-phosphate-based nano material. The algae were incubated in media with a sublethal zinc concentration (6.53 mg Zn dm^-3) for 4 weeks. Using BF-and ADF-STEM imaging combined with analytical XEDS we could show that nano needles containing phosphorus and zinc were formed inside the living cells. Further more, the cells incubated with zinc show a strong fluorescence. Our findings indicate that the algae used polyphosphate bodies for detoxification of the zinc ions, leading to the generation of intracellular zinc-phosphate-based nano needles. Beside the technological application of this material, the fluorescent cells can be used for labeling of e.g. biological probes. This new experimental protocol for the production of an inorganic functional material can be applied also for other substances

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Extended abstract of a paper presented at Microscopy and Microanalysis 2011 in Nashville, Tennessee, USA, August 7–August 11, 2011.

Extended abstract of a paper presented at Microscopy and Microanalysis 2013 in Indianapolis, Indiana, USA, August 4 – August 8, 2013.