Functional adaptations in the tergite cuticle of the desert isopod Hemilepistus reaumuri (Milne-Edwards, 1840).

To survive in its extreme habitat, the cuticle of the burrowing desert isopod Hemilepistus reaumuri requires properties distinct from isopods living in moist or mesic habitats. In particular, the anterior tergites are exposed to high mechanical loads and temperatures when individuals guard the entrance of their burrow. We have, therefore, investigated the architecture, composition, calcite texture and local mechanical properties of the tergite cuticle, with particular emphasis on large anterior cuticle tubercles and differences between the anterior and posterior tergite. Unexpectedly, structure and thickness of the epicuticle resemble those in mesic isopod species. The anterior tergite has a thicker endocuticle and a higher local stiffness than the posterior tergite. Calcite distribution in the cuticle is unusual, because in addition to the exocuticle the endocuticle distally also contains calcite. The calcite consists of a distal layer of dense and highly co-oriented crystal-units, followed proximally by irregularly distributed and, with respect to each other, misoriented calcite crystallites. The calcite layer at the tip of the tubercle is thicker relative to the tubercle slopes, and its crystallites are more misoriented to each other. A steep decrease of local stiffness and hardness is observed within a distal region of the cuticle, likely caused by a successive increase in the ACC/calcite ratio rather than changes in the degree of mineralisation. Comparison of the results with other isopods reveals a much lower ACC/calcite ratio in H. reaumuri and a correlation between the degree of terrestriality of isopod species and the magnesium content of the cuticle.

Tailored disorder in calcite organization in tergite cuticle of the supralittoral isopod Tylos europaeus Arcangeli, 1938.

The crustacean cuticle forms skeletal elements consisting of chitin-protein fibrils reinforced by amorphous and crystalline calcium carbonate and phosphate minerals. The edges of skeletal elements are of particular interest. They are subject to repeated strain and stress because they form transitions to the arthrodial membranes connecting them. These allow for relative movements of skeletal elements. In this study, we investigate structure, chemical composition, mineral organization and local mechanical properties of the anterior and posterior edges of the tergite cuticle in the conglobating beach isopod Tylos europaeus and compare these with the protective dorsal region of the tergites. The distribution of mineral phases at the edges resembles that of dorsal regions of the tergites. At the transition with the unmineralized arthrodial membrane the calcite containing distal exocuticle is replaced by epicuticular material and the subjacent cuticular layers containing amorphous calcium carbonate become enriched with amorphous calcium phosphate. At the edges, the local elastic modulus and hardness values are significantly lower compared to dorsal regions of the tergite cuticle, for both, the calcite and the amorphous mineral containing layers. The calcite within the tergite cuticle is assembled in different texture patterns: (i) almost random co-orientation, (ii) almost single crystalline calcite, and (iii) a graded organization. Calcite organization and co-orientation strength is highly variable, not only on very few tens of micrometres, but also between regions with different skeletal functionality. Our results show that besides structure and composition, patterns of calcite organization contribute to the hierarchical architecture and functionality of biological composites.

Function-related adaptations of ultrastructure, mineral phase distribution and mechanical properties in the incisive cuticle of mandibles of Porcellio scaber Latreille, 1804.

In terrestrial isopods the mandibles consist of a corpus carrying strong muscle tissue, and a pars incisiva (PI) that cuts dry leaves into smaller ingestible pieces. We studied the cuticle of the PI of Porcellio scaber in order to understand region-dependent differences in its ultrastructure, composition, and the resulting mechanical properties, employing several microscopic and analytical techniques as well as nanoindentation experiments. The cuticle of the incisive tip is not mineralized and consists of an unusually thick epicuticle containing thin fibrils, two subjacent cuticular layers, and a central core containing fibrils of different orientation, either longitudinal or circumferential. A thick epicuticle of the middle region just behind the tip projects long epicuticular extensions into the subjacent endocuticle, likely to prevent delamination. A distinct exocuticular layer is lacking in the middle region. Most chitin-protein fibrils within the endocuticle are oriented in parallel pointing towards the tip. Surprisingly, the middle region is mineralized by amorphous calcium phosphate (ACP) only. Near the base, ACP is successively replaced by amorphous calcium carbonate and calcite is restricted to a distal layer in the base. At the transition between middle and base, the epicuticle forms a hybrid material containing fibrils of the exocuticle. Nanoindentation experiments reveal an increase of the stiffness and hardness from the tip towards the base and significantly higher values on transversal in comparison to longitudinal planes. The results suggest that ultrastructure and composition are adapted for conveying high forces from a rather thin cutting edge to the stable base of the PI.

Characterization of crocodile teeth: correlation of composition, microstructure, and hardness.

Structure and composition of teeth of the saltwater crocodile Crocodylus porosus were characterized by several high-resolution analytical techniques. X-ray diffraction in combination with elemental analysis and infrared spectroscopy showed that the mineral phase of the teeth is a carbonated calcium-deficient nanocrystalline hydroxyapatite in all three tooth-constituting tissues: Dentin, enamel, and cementum. The fluoride content in the three tissues is very low (<0.1 wt.%) and comparable to that in human teeth. The mineral content of dentin, enamel, and cementum as determined by thermogravimetry is 71.3, 80.5, and 66.8 wt.%, respectively. Synchrotron X-ray microtomography showed the internal structure and allowed to visualize the degree of mineralization in dentin, enamel, and cementum. Virtual sections through the tooth and scanning electron micrographs showed that the enamel layer is comparably thin (100-200 µm). The crystallites in the enamel are oriented perpendicularly to the tooth surface. At the dentin-enamel-junction, the packing density of crystallites decreases, and the crystallites do not display an ordered structure as in the enamel. The microhardness was 0.60±0.05 GPa for dentin, 3.15±0.15 GPa for enamel, 0.26±0.08 GPa for cementum close to the crown, and 0.31±0.04 GPa for cementum close to the root margin. This can be explained with the different degree of mineralization of the different tissue types and is comparable with human teeth.

Toothpaste Abrasion and Abrasive Particle Content: Correlating High-Resolution Profilometric Analysis with Relative Dentin Abrasivity (RDA).

In this in vitro study, the influence of the concentration of abrasive particles on the abrasivity of toothpastes was investigated using laser scan profilometry on polymethyl methacrylate (PMMA) surfaces with the aim of providing an alternative method to developers for screening of new toothpaste formulations. PMMA plates were tested in a toothbrush simulator with distilled water and four model toothpastes with increasing content of hydrated silica (2.5, 5.0, 7.5, 10.0 wt%). The viscosity of the model toothpaste formulations was kept constant by means of varying the content of sodium carboxymethyl cellulose and water. The brushed surfaces were evaluated using laser scan profilometry at micrometer-scale resolutions, and the total volume of the introduced scratches was calculated along with the roughness parameters Ra, Rz and Rv. RDA measurements commissioned for the same toothpaste formulations were used to analyze the correlation between results obtained with the different methods. The same experimental procedure was applied to five commercially available toothpastes, and the results were evaluated against our model system. In addition, we characterize abrasive hydrated silica and discuss their effects on PMMA-sample surfaces. The results show that the abrasiveness of a model toothpaste increases with the weight percentage of hydrated silica. Increasing roughness parameter and volume loss values show good correlation with the likewise increasing corresponding RDA values for all model toothpastes, as well as commercial toothpastes without ingredients that can damage the used substrate PMMA. From our results, we deduce an abrasion classification that corresponds to the RDA classification established for marketed toothpastes.

Overview of Calcium Phosphates used in Biomimetic Oral Care.

BACKGROUND: The use of biomimetic agents is an emerging field in modern oral care. Promising biomimetic substances for such applications are calcium phosphates, because their chemical composition is very similar to that of the mineral phase in human teeth, especially of natural enamel. Examples for their application include the remineralization of early caries lesions and repair of small enamel defects. OBJECTIVE: This review provides an interdisciplinary view on calcium phosphates and their applications in biomimetic oral care. The aim of this work is to give an overview of in vivo and in situ studies comparing several calcium phosphates in preventive dentistry that can be used as a knowledge base for the development of innovative alternative oral care concepts. METHODS: Books, reviews, and original research papers with a focus on in vivo and in situ studies were included. The databases PubMed® and SciFinder® were used for literature search. Calcium phosphates that are frequently utilized in oral care products are covered in this review and were used as search terms alone and together with the following key words: in vivo, in situ, caries, clinical study, and remineralization. From 13,470 studies found, 35 studies complied with the inclusion criteria and were used for this review. RESULTS: Published in vivo and in situ studies demonstrate calcium phosphates' potential in enamel remineralization. However, more studies are needed to further substantiate existing results and to extend and refine the application of calcium phosphates in modern oral care. CONCLUSION: Calcium phosphates represent an innovative biomimetic approach for daily oral care because of their high similarity to natural enamel that will broaden the range of future treatments in preventive dentistry.

From insect scales to sensor design: modelling the mechanochromic properties of bicontinuous cubic structures.

Many of the three-dimensional photonic crystals occurring in the scales of insects have bicontinuous cubic structures. Their optical properties have been studied extensively, however little is known about their mechanical properties and their optical response under deformation. We demonstrated a mechanochromic effect by deforming the scales of a weevil and calculated the elastic, optical and mechanochromic (assuming homogeneous deformation) properties of the three types of bicontinuous cubic structures occurring in nature: P-structure (primitive), G-structure (gyroid) and D-structure (diamond). The results show that all investigated properties of these three structure types strongly depend on their geometry, structural parameters such as volume fractions of the two constituting phases and the directions of the incident light or applied stress, respectively. Interestingly, the mechanochromic simulation results predict that these structures may show blue-shift or even red-shift under compression along certain directions. Our results provide design guidelines for mechanochromic sensing materials operating in the elastic regime, including parameters such as sensitivity and direction of spectral shift.

Composites of fluoroapatite and methylmethacrylate-based polymers (PMMA) for biomimetic tooth replacement.

Synthetic composite materials that mimic the structure and composition of mammalian tooth enamel were prepared by mixing fluoroapatite rods (diameter 2-3 µm, thickness about 0.5 µm) and methylmethacrylate (MMA), followed by polymerization either during or immediately after ultracentrifugation, using either a tertiary amine/radical initiator for polymerization at room temperature or a radical initiator for thermal polymerization. This led to mineral-rich composites (mineral content between 50 and 75 wt%). To enhance the mechanical stability and the interaction between fluoroapatite and polymer matrix, small amounts of differently functionalized MMA monomers were added to the co-monomer mixture. Another approach was the coating of the fluoroapatite rods with silica and the polymerization in the presence of a siloxane-functionalized MMA monomer. The hardness of the composites was about 0.2-0.4 GPa as determined by Vickers indentation tests, about 2 times higher than the polymer matrix alone. The composites had a good resistance against acids (60 min at pH 3, 37 °C).

Functional adaptation of crustacean exoskeletal elements through structural and compositional diversity: a combined experimental and theoretical study.

The crustacean cuticle is a composite material that covers the whole animal and forms the continuous exoskeleton. Nano-fibers composed of chitin and protein molecules form most of the organic matrix of the cuticle that, at the macroscale, is organized in up to eight hierarchical levels. At least two of them, the exo- and endocuticle, contain a mineral phase of mainly Mg-calcite, amorphous calcium carbonate and phosphate. The high number of hierarchical levels and the compositional diversity provide a high degree of freedom for varying the physical, in particular mechanical, properties of the material. This makes the cuticle a versatile material ideally suited to form a variety of skeletal elements that are adapted to different functions and the eco-physiological strains of individual species. This review presents our recent analytical, experimental and theoretical studies on the cuticle, summarising at which hierarchical levels structure and composition are modified to achieve the required physical properties. We describe our multi-scale hierarchical modeling approach based on the results from these studies, aiming at systematically predicting the structure-composition-property relations of cuticle composites from the molecular level to the macro-scale. This modeling approach provides a tool to facilitate the development of optimized biomimetic materials within a knowledge-based design approach.

Ultrastructural organization and micromechanical properties of shark tooth enameloid.

The outer part of shark teeth is formed by the hard and mineral-rich enameloid that has excellent mechanical properties, which makes it a very interesting model system for the development of new bio-inspired dental materials. We characterized the microstructure, chemical composition and resulting local mechanical properties of the enameloid from teeth of Isurus oxyrinchus (shortfin mako shark) by performing an in-depth analysis using various high-resolution analytical techniques, including scanning electron microscopy, qualitative energy-dispersive X-ray spectroscopy and nanoindentation. Shark tooth enameloid reveals an intricate hierarchical arrangement of thin (50-80nm) and long (>1µm) crystallites of fluoroapatite with a high degree of structural anisotropy, which leads to exceptional mechanical properties. Both stiffness and hardness are surprisingly homogeneous in the shiny layer as well as in the enameloid: although both tooth phases differ in structure and composition, they show almost no orientation dependence with respect to the loading direction of the enameloid crystallites. The results were used to determine the structural hierarchy of shark teeth, which can be used as a base for establishing design criteria for synthetic bio-inspired and biomimetic dental composites.

Ab initio study of thermodynamic, structural, and elastic properties of Mg-substituted crystalline calcite.

Arthropoda, which represent nearly 80% of all known animal species, are protected by an exoskeleton formed by their cuticle. The cuticle represents a hierarchically structured multifunctional biocomposite based on chitin and proteins. Some groups, such as Crustacea, reinforce the load-bearing parts of their cuticle with calcite. As the calcite sometimes contains Mg it was speculated that Mg may have a stiffening impact on the mechanical properties of the cuticle (Becker et al., Dalton Trans. (2005) 1814). Motivated by these facts, we present a theoretical parameter-free quantum-mechanical study of the phase stability and structural and elastic properties of Mg-substituted calcite crystals. The Mg-substitutions were chosen as examples of states that occur in complex chemical environments typical for biological systems in which calcite crystals contain impurities, the role of which is still the topic of debate. Density functional theory calculations of bulk (Ca,Mg)CO3 were performed employing 30-atom supercells within the generalized gradient approximation as implemented in the Vienna Ab-initio Simulation Package. Based on the calculated thermodynamic results, low concentrations of Mg atoms are predicted to be stable in calcite crystals in agreement with experimental findings. Examining the structural characteristics, Mg additions nearly linearly reduce the volume of substituted crystals. The predicted elastic bulk modulus results reveal that the Mg substitution nearly linearly stiffens the calcite crystals. Due to the quite large size-mismatch of Mg and Ca atoms, Mg substitution results in local distortions such as off-planar tilting of the CO3²â» group.