Biomineral electron backscatter diffraction for palaeontology

Electron backscatter diffraction (EBSD) originated in materials science and has transferred to biomineral research providing insight into fossil and modern biominerals. An electron microscopy technique, EBSD requires a fine polished sample surface where the electron beam diffracts in the first few lattice layers, identifying mineral, polymorph and crystallographic orientation. The technique is particularly well suited for the analysis of modern and fossil calcium carbonate biominerals, where it provides key insight into biological control of mineral formation such as in molluscs and brachiopods. EBSD readily identifies original and secondary mineralogy, which helps to inform our understanding of biomineral evolution such as the identification of original aragonite in Silurian trimerellid brachiopods. As a technique to identify and thus avoid the inclusion of secondary minerals in proxy organisms such as corals, EBSD can be used to ensure accuracy of palaeoproxy data. Even when fossil systems have no modern equivalents, EBSD can provide key data to determine functional mechanisms such as in the lenses of schizochroal eyes of phacopine trilobites. These few examples illustrate that EBSD is proving to be a valuable component of the palaeontology toolkit

Assessment of crystallographic influence on material properties of calcite brachiopods

Calcium carbonate biominerals are frequently analysed in materials science due to their abundance, diversity and unique material properties. Aragonite nacre is intensively studied, but less information is available about the material properties of biogenic calcite, despite its occurrence in a wide range of structures in different organisms. In particular, there is insufficient knowledge about how preferential crystallographic orientations influence these material properties. Here, we study the influence of crystallography on material properties in calcite semi-nacre and fibres of brachiopod shells using nano-indentation and electron backscatter diffraction (EBSD). The nano-indentation results show that calcite semi-nacre is a harder and stiffer (H {approx} 3–5 GPa; E = 50–85 GPa) biomineral structure than calcite fibres (H = 0.4–3 GPa; E = 30–60 GPa). The integration of EBSD to these studies has revealed a relationship between the crystallography and material properties at high spatial resolution for calcite semi-nacre. The presence of crystals with the c-axis perpendicular to the plane-of-view in longitudinal section increases hardness and stiffness. The present study determines how nano-indentation and EBSD can be combined to provide a detailed understanding of biomineral structures and their analysis for application in materials science

Transfer of a large gene regulatory apparatus to a new developmental address in echinoid evolution

Of the five echinoderm classes, only the modern sea urchins (euechinoids) generate a precociously specified embryonic micromere lineage that ingresses before gastrulation and then secretes the biomineral embryonic skeleton. The gene regulatory network (GRN) underlying the specification and differentiation of this lineage is now known. Many of the same differentiation genes as are used in the biomineralization of the embryo skeleton are also used to make the similar biomineral of the spines and test plates of the adult body. Here, we determine the components of the regulatory state upstream of these differentiation genes that are shared between embryonic and adult skeletogenesis. An abrupt “break point” in the micromere GRN is thus revealed, on one side of which most of the regulatory genes are used in both, and on the other side of which the regulatory apparatus is entirely micromere-specific. This reveals the specific linkages of the micromere GRN forged in the evolutionary process by which the skeletogenic gene batteries were caused to be activated in the embryonic micromere lineage. We also show, by comparison with adult skeletogenesis in the sea star, a distant echinoderm outgroup, that the regulatory apparatus responsible for driving the skeletogenic differentiation gene batteries is an ancient pleisiomorphic aspect of the echinoderm-specific regulatory heritage

Biomineral repair of Abalone shell apertures

The shell of the gastropod mollusc, abalone, is comprised of nacre with an outer prismatic layer that is composed of either calcite or aragonite or both, depending on the species. A striking characteristic of the abalone shell is the row of apertures along the dorsal margin. As the organism and shell grow, new apertures are formed and the preceding ones are filled in. Detailed investigations, using electron backscatter diffraction, of the infill in three species of abalone: Haliotis asinina, Haliotis gigantea and Haliotis rufescens reveals that, like the shell, the infill is composed mainly of nacre with an outer prismatic layer. The infill prismatic layer has identical mineralogy as the original shell prismatic layer. In H. asinina and H. gigantea, the prismatic layer of the shell and infill are made of aragonite while in H. rufescens both are composed of calcite. Abalone builds the infill material with the same high level of biological control, replicating the structure, mineralogy and crystallographic orientation as for the shell. The infill of abalone apertures presents us with insight into what is, effectively, shell repair

Development of human limbs.

This work offers a new view on the developmental history of tetrapods. It proposes an original evolution model of human limbs based on metameric formation of osteogenic buds in accordance to primary segmentation and biplanar symmetry. While going through evolution, osteogenic buds initially identical to each other were changing their sizes, realigning, regressing, uniting while keeping the direction of the formation in accordance to the following formula (taking into account sesamoid bones):
2; 1; 2; 3; 2; 3; 5; 5; 8; 8 (in the upper limb together with the upper limb girdle); 3; 2; 3; 2; 1; 2; 8; 8; 5; 5 (in the lower limb together with the pelvic bones)

Biological niches within human calcified aortic valves. Towards understanding of the pathological biomineralization process

Despite recent advances, mineralization site, its microarchitecture, and composition in calcific heart valve remain poorly understood. A multiscale investigation, using scanning electron microscopy (SEM), transmission electron microscopy (TEM), and energy dispersive X-ray spectrometry (EDS), from micrometre up to nanometre, was conducted on human severely calcified aortic and mitral valves, to provide new insights into calcificationp rocess. Our aim was to evaluate the spatial relationship existing between bioapatite crystals, their local growing microenvironment, and the presence of a hierarchical architecture. Here we detected the presence of bioapatite crystals in two different mineralization sites that suggest the action of two different growth processes:a pathological crystallization process that occurs in biological niches and is ascribed to a purely physicochemical process and a matrix- mediated mineralized process in which the extracellular matrix acts as the template for a site-directed nanocrystals nucleation. Different shapes of bioapatite crystallization were observed at micrometer scale in each microenvironment but at the nanoscale level crystals appear to be made up by the same subunit

Biominerals - source and inspiration for novel advanced materials

Biomineralization seems an odd sort of word. How can you combine biology and minerals? However, a quick look around brings to light many familiar objects that are examples of biominerals. Most dramatic are the coral reefs and sea shells of the marine environment (calcium carbonate) and human bone and teeth (calcium hydroxyapatite) but there are many other examples. In the past 10 years, an increasing number of biominerals has been reported (Table 1). Interest in the biological and chemical processes that lead to biomineralization, howeyer, has only developed rather recently. Early observations were made by paleontologists who were interested in the preservation, through geological time, of the hard parts of organisms such as shells and skeletons but only in 1989 did the field really come of age with the almost simultaneous publication of three monographs covering current knowledge of the biological, biochemical, chemical and taxonomic aspects of biomineralization (Mann et al. 1989; Lowenstam & Weiner 1989; Simkiss & Wilbur 1989)

Toward a Structure Determination Method for Biomineral-Associated Protein Using Combined Solid- State NMR and Computational Structure Prediction

SummaryProtein-biomineral interactions are paramount to materials production in biology, including the mineral phase of hard tissue. Unfortunately, the structure of biomineral-associated proteins cannot be determined by X-ray crystallography or solution nuclear magnetic resonance (NMR). Here we report a method for determining the structure of biomineral-associated proteins. The method combines solid-state NMR (ssNMR) and ssNMR-biased computational structure prediction. In addition, the algorithm is able to identify lattice geometries most compatible with ssNMR constraints, representing a quantitative, novel method for investigating crystal-face binding specificity. We use this method to determine most of the structure of human salivary statherin interacting with the mineral phase of tooth enamel. Computation and experiment converge on an ensemble of related structures and identify preferential binding at three crystal surfaces. The work represents a significant advance toward determining structure of biomineral-adsorbed protein using experimentally biased structure prediction. This method is generally applicable to proteins that can be chemically synthesized