Igor Zlotnikov

Self-similar mesostructure evolution of the growing mollusc shell reminiscent of thermodynamically driven grain growth

Significant progress has been made in understanding the interaction between mineral precursors and organic components leading to material formation and structuring in biomineralizing systems1–5. The mesostructure of biological materials, such as the outer calcitic shell of molluscs, is characterized by many parameters and the question arises as to what extent they all are, or need to be, controlled biologically. Here, we analyse the three-dimensional structure of the calcite-based prismatic layer ofPinna nobilis6–8, the giant Mediterranean fan mussel, using high-resolution synchrotronbased microtomography. We show that the evolution of the layer is statistically self-similar and, remarkably, its morphology and mesostructure can be fully predicted using classicalmaterials science theories for normal grain growth9–16. These findings are a fundamental step in understanding the constraints that dictate the shape of these biogenic minerals and shed light on how biological organisms make use of thermodynamics to generate complex morphologies.

A perfectly periodic three-dimensional protein/silica mesoporous structure produced by an organism.

The discovery of perfectly ordered 3D mesoporous protein/silica structure in the axial filament of the marine sponge Monorhaphis chuni is reported. The structure belongs to body-centered tetragonal symmetry system (a=9.88 nm, c=10.83 nm) and comprises interconnecting lattices of protein and silica, templated by the self-assembled, enzymatically active protein-silicatein, whose primary function is the precipitation of silica.

In situ elastic modulus measurements of ultrathin protein-rich organic layers in biosilica: towards deeper understanding of superior resistance to fracture of biocomposites

Biogenic ceramics are known to exhibit superior toughness due to a laminated architecture with ultrathin organic layers separating the ceramic blocks. Theoretical analyses relate the toughness increase to the modulus contrast, Ec/Eo between the stiff, Ec, and the compliant, Eo, components. However, experimental data on this contrast are extremely difficult to obtain by any known technique due to the very small thickness and low modulus values of the organic layers. Here we adapt a recently developed nanoscale modulus mapping technique combined with reverse finite element analysis in order to map the elastic modulus across a 35 nm thick organic layer within biosilica in a giant anchor spicule of the glass sponge Monorhaphis chuni. We find a modulus of 0.7 GPa in the organic layer as compared to 37 GPa in the bioglass. Furthermore, a modulus gradient extends 50 nm into the glass layer, probably due to the spatial distribution of small organic inclusions. With this new methodology it becomes possible to determine the elastic moduli of nanometric inclusions even when embedded in a matrix which is 50 times stiffer.

Structural and mechanical properties of the arthropod cuticle: Comparison between the fang of the spider Cupiennius salei and the carapace of American lobster Homarus americanus

Most biological materials are nanocomposites characterized by a multi-level structural hierarchy. Particularly, the arthropod cuticle is a chitin-based composite material where the mechanical properties strongly depend on both molecular chitin/protein properties, and the structural arrangement of chitin-fibrils within the protein matrix. Here materials properties and structural organization of two types of cuticle from distantly related arthropods, the wandering spider Cupiennius salei and American lobster Homarus americanus were studied using nanoindentation and X-ray diffraction. The structural analysis of the two types of cuticle including the packing and alignment of chitin-fibrils is supported by Monte Carlo simulations of the experimental X-ray data, thereby regions of parallel and rotated fibril arrangement can be clearly distinguished. The tip of the spider fang which is used to inject venom into the prey was found to be considerably harder than the lobster carapace, while its stiffness is slightly lower.

A spider's biological vibration filter: Micromechanical characteristics of a biomaterial surface

A strain-sensing lyriform organ (HS-10) found on all of the legs of a Central American wandering spider (Cupiennius salei) detects courtship, prey and predator vibrations transmitted by the plant on which it sits. It has been suggested that the viscoelastic properties of a cuticular pad directly adjacent to the sensory organ contribute to the organs pronounced high-pass characteristics. Here, we investigate the micromechanical properties of the cuticular pad biomaterial in search of a deeper understanding of its impact on the function of the vibration sensor. These properties are considered to be an effective adaptation for the selective detection of signals for frequencies >40 Hz. Using surface force spectroscopy mapping we determine the elastic modulus of the pad surface over a temperature range of 15-40 °C at various loading frequencies. In the glassy state, the elastic modulus was ~100 MPa, while in the rubbery state the elastic modulus decreased to 20 MPa. These data are analyzed according to the principle of time-temperature superposition to construct a master curve that relates mechanical properties, temperature and stimulus frequencies. By estimating the loss and storage moduli vs. temperature and frequency it was possible to make a direct comparison with electrophysiology experiments, and it was found that the dissipation of energy occurs within a frequency window whose position is controlled by environmental temperatures.

Hierarchical Structuring of Liquid Crystal Polymer-Laponite Hybrid Materials

Biomimetic organic-inorganic composite materials were fabricated via one-step self-organization on three hierarchical levels. The organic component was a polyoxazoline with pendent cholesteryl and carboxyl (N-Boc-protected amino acid) side chains that was able to form a chiral nematic lyotropic phase and bind to positively charged inorganic faces of Laponite. The Laponite particles formed a mesocrystalline arrangement within the liquid-crystal (LC) polymer phase upon shearing a viscous dispersion of Laponite nanoparticles and LC polymer in DMF. Complementary analytical and mechanical characterization techniques (AUC, POM, TEM, SEM, SAXS, μCT, and nanoindentation) covering the millimeter, micrometer, and nanometer length scales reveal the hierarchical structures and properties of the composite materials consisting of different ratios of Laponite nanoparticles and liquid-crystalline polymer.

Micro- and nano-structural details of a spider's filter for substrate vibrations: relevance for low-frequency signal transmission

The metatarsal lyriform organ of the Central American wandering spider Cupiennius salei is its most sensitive vibration detector. It is able to sense a wide range of vibration stimuli over four orders of magnitude in frequency between at least as low as 0.1 Hz and several kilohertz. Transmission of the vibrations to the slit organ is controlled by a cuticular pad in front of it. While the mechanism of high-frequency stimulus transfer (above ca 40 Hz) is well understood and related to the viscoelastic properties of the pads epicuticle, it is not yet clear how low-frequency stimuli (less than 40 Hz) are transmitted. Here, we study how the pad material affects the pads mechanical properties and thus its role in the transfer of the stimulus, using a variety of experimental techniques, such as X-ray micro-computed tomography for three-dimensional imaging, X-ray scattering for structural analysis, and atomic force microscopy and scanning electron microscopy for surface imaging. The mechanical properties were investigated using scanning acoustic microscopy and nanoindentation. We show that large tarsal deflections cause large deformation in the distal highly hydrated part of the pad. Beyond this region, a sclerotized region serves as a supporting frame which resists the deformation and is displaced to push against the slits, with displacement values considerably scaled down to only a few micrometres. Unravelling the structural arrangement in such specialized structures may provide conceptual ideas for the design of new materials capable of controlling a technical sensors specificity and selectivity, which is so typical of biological sensors.

Eshelby Twist as a Possible Source of Lattice Rotation in a Perfectly Ordered Protein/Silica Structure Grown by a Simple Organism

The formation mechanism of a perfectly ordered protein/silica structure in the axial filament of the anchor spicule of the silica sponge Monorhaphis chuni is suggested. Experimental evidence shows that the growth of this architecture is realized by a thermodynamically driven dislocation-mediated spiral growth mechanism, resulting in a specific rotation of the mesoscopic crystal lattice (Eshelby twist).

Characterizing moisture-dependent mechanical properties of organic materials: humidity-controlled static and dynamic nanoindentation of wood cell walls

Nanoindentation is an ideal technique to study local mechanical properties of a wide range of materials on the sub-micron scale. It has been widely used to investigate biological materials in the dry state; however, their properties are strongly affected by their moisture content, which until now has not been consistently controlled. In the present study, we developed an experimental set-up for measuring local mechanical properties of materials by nanoindentation in a controlled environment of relative humidity (RH) and temperature. The significance of this new approach in studying biological materials was demonstrated for the secondary cell wall layer (S2) in Spruce wood (Picea abies). The hardness of the cell wall layer decreased from an average of approximately 0.6 GPa at 6% RH down to approximately 0.2 GPa at 79% RH, corresponding to a reduction by a factor of 3. Under the same conditions, the indentation modulus also decreased by about 40%. The newly designed experimental set-up has a strong potential for a variety of applications involving the temperature- and humidity-dependent properties of biological and artificial organic nanocomposites.

Gas barrier properties of bio-inspired Laponite–LC polymer hybrid films

Bio-inspired Laponite (clay)-liquid crystal (LC) polymer composite materials with high clay fractions (>80%) and a high level of orientation of the clay platelets, i.e. with structural features similar to the ones found in natural nacre, have been shown to exhibit a promising behavior in the context of reduced oxygen transmission. Key characteristics of these bio-inspired composite materials are their high inorganic content, high level of exfoliation and orientation of the clay platelets, and the use of a LC polymer forming the organic matrix in between the Laponite particles. Each single feature may be beneficial to increase the materials gas barrier property rendering this composite a promising system with advantageous barrier capacities. In this detailed study, Laponite/LC polymer composite coatings with different clay loadings were investigated regarding their oxygen transmission rate. The obtained gas barrier performance was linked to the quality, respective Laponite content and the underlying composite micro- and nanostructure of the coatings. Most efficient oxygen barrier properties were observed for composite coatings with 83% Laponite loading that exhibit a structure similar to sheet-like nacre. Further on, advantageous mechanical properties of these Laponite/LC polymer composites reported previously give rise to a multifunctional composite system.