Calcitic Prisms of The Giant Seashell Pinna Nobilis Form Light Guide Arrays.

The shells of the Pinnidae family are based on a double layer of single-crystal-like calcitic prisms and inner aragonitic nacre, a structure known for its outstanding mechanical performance. However, on the posterior side, shells are missing the nacreous layer, which raises the question of whether there can be any functional role in giving up this mechanical performance. Here, it is demonstrated that the prismatic part of the Pinna nobilis shell exhibits unusual optical properties, whereby each prism acts as an individual optical fiber guiding the ambient light to the inner shell cavity by total internal reflection. This pixelated light channeling enhances both spatial resolution and contrast while reducing angular blurring, an apt combination for acute tracking of a moving object. These findings offer insights into the evolutionary aspects of light-sensing and imaging and demonstrate how an architectured optical system for efficient light-tracking can be based on birefringent ceramics.

Hydrogels with stiffness-degradation spatial patterns control anisotropic 3D cell response.

In nature, tissues are patterned, but most biomaterials used in human applications are not. Patterned biomaterials offer the opportunity to mimic spatially segregating biophysical and biochemical properties found in nature. Engineering such properties allows to study cell-matrix interactions in anisotropic matrices in great detail. Here, we developed alginate-based hydrogels with patterns in stiffness and degradation, composed of distinct areas of soft non-degradable (Soft-NoDeg) and stiff degradable (Stiff-Deg) material properties. The hydrogels exhibit emerging patterns in stiffness and degradability over time, taking advantage of dual crosslinking: Diels-Alder covalent crosslinking (norbornene-tetrazine, non degradable) and UV-mediated peptide crosslinking (matrix metalloprotease sensitive peptide, enzymatically degradable). The materials were mechanically characterized using rheology for single-phase and surface micro-indentation for patterned materials. 3D encapsulated mouse embryonic fibroblasts (MEFs) allowed to characterize the anisotropic cell-matrix interaction in terms of cell morphology by employing a novel image-based quantification tool. Live/dead staining showed no differences in cell viability but distinct patterns in proliferation, with higher cell number in Stiff-Deg materials at day 14. Patterns of projected cell area became visible already at day 1, with larger values in Soft-NoDeg materials. This was inverted at day 14, when larger projected cell areas were identified in Stiff-Deg. This shift was accompanied by a significant decrease in cell circularity in Stiff-Deg. The control of anisotropic cell morphology by the material patterns was also confirmed by a significant increase in filopodia number and length in Stiff-Deg materials. The novel image-based quantification tool was useful to spatially visualize and quantify the anisotropic cell response in 3D hydrogels with stiffness-degradation spatial patterns. Our results show that patterning of stiffness and degradability allows to control cell anisotropic response in 3D and can be quantified by image-based strategies. This allows a deeper understanding of cell-matrix interactions in a multicomponent material.

Crystal orientation mapping and microindentation reveal anisotropy in Porites skeletons.

Structures made by scleractinian corals support diverse ocean ecosystems. Despite the importance of coral skeletons and their predicted vulnerability to climate change, few studies have examined the mechanical and crystallographic properties of coral skeletons at the micro- and nano-scales. Here, we investigated the interplay of crystallographic and microarchitectural organization with mechanical anisotropy within Porites skeletons by measuring Young's modulus and hardness along surfaces transverse and longitudinal to the primary coral growth direction. We observed micro-scale anisotropy, where the transverse surface had greater Young's modulus and hardness by ∼ 6 GPa and 0.2 GPa, respectively. Electron backscatter diffraction (EBSD) revealed that this surface also had a higher percentage of crystals oriented with the a-axis between ± 30-60∘, relative to the longitudinal surface, and a broader grain size distribution. Within a region containing a sharp microscale gradient in Young's modulus, nanoscale indentation mapping, energy dispersive spectroscopy (EDS), EBSD, and Raman crystallography were performed. A correlative trend showed higher Young's modulus and hardness in regions with individual crystal bases (c-axis) facing upward, and in crystal fibers relative to centers of calcification. These relationships highlight the difference in mechanical properties between scales (i.e. crystals, crystal bundles, grains). Observations of crystal orientation and mechanical properties suggest that anisotropy is driven by microscale organization and crystal packing rather than intrinsic crystal anisotropy. In comparison with previous observations of nanoscale isotropy in corals, our results illustrate the role of hierarchical architecture in coral skeletons and the influence of biotic and abiotic factors on mechanical properties at different scales. STATEMENT OF SIGNIFICANCE: Coral biomineralization and the ability of corals' skeletal structure to withstand biotic and abiotic forces underpins the success of reef ecosystems. At the microscale, we show increased skeletal stiffness and hardness perpendicular to the coral growth direction. By comparing nano- and micro-scale indentation results, we also reveal an effect of hierarchical architecture on the mechanical properties of coral skeletons and hypothesize that crystal packing and orientation result in microscale anisotropy. In contrast to previous findings, we demonstrate that mechanical and crystallographic properties of coral skeletons can vary between surface planes, within surface planes, and at different analytical scales. These results improve our understanding of biomineralization and the effects of scale and direction on how biomineral structures respond to environmental stimuli.

Interplay between Interfacial Energy, Contact Mechanics, and Capillary Forces in EGaIn Droplets.

Eutectic gallium-indium (EGaIn) is increasingly employed as an interfacial conductor material in molecular electronics and wearable healthcare devices owing to its ability to be shaped at room temperature, conductivity, and mechanical stability. Despite this emerging usage, the mechanical and physical mechanisms governing EGaIn interactions with surrounding objects─mainly regulated by surface tension and interfacial adhesion─remain poorly understood. Here, using depth-sensing nanoindentation (DSN) on pristine EGaIn/GaOx surfaces, we uncover how changes in EGaIn/substrate interfacial energies regulate the adhesive and contact mechanic behaviors, notably the evolution of EGaIn capillary bridges with distinct capillary geometries and pressures. Varying the interfacial energy by subjecting EGaIn to different chemical environments and by functionalizing the tip with chemically distinct self-assembled monolayers (SAMs), we show that the adhesion forces between EGaIn and the solid substrate can be increased by up to 2 orders of magnitude, resulting in about a 60-fold increase in the elongation of capillary bridges. Our data reveal that by deploying molecular junctions with SAMs of different terminal groups, the trends of charge transport rates, the resistance of monolayers, and the contact interactions between EGaIn and monolayers from electrical characterizations are governed by the interfacial energies as well. This study provides a key understanding into the role of interfacial energy on geometrical characteristics of EGaIn capillary bridges, offering insights toward the fabrication of EGaIn junctions in a controlled fashion.

Groovy and Gnarly: Surface Wrinkles as a Multifunctional Motif for Terrestrial and Marine Environments.

From large ventral pleats of humpback whales to nanoscale ridges on flower petals, wrinkled structures are omnipresent, multifunctional, and found at hugely diverse scales. Depending on the particulars of the biological system-its environment, morphology, and mechanical properties-wrinkles may control adhesion, friction, wetting, or drag; promote interfacial exchange; act as flow channels; or contribute to stretching, mechanical integrity, or structural color. Undulations on natural surfaces primarily arise from stress-induced instabilities of surface layers (e.g., buckling) during growth or aging. Variation in the material properties of surface layers and in the magnitude and orientation of intrinsic stresses during growth lead to a variety of wrinkling morphologies and patterns which, in turn, reflect the wide range of biophysical challenges wrinkled surfaces can solve. Therefore, investigating how surface wrinkles vary and are implemented across biological systems is key to understanding their structure-function relationships. In this work, we synthesize the literature in a metadata analysis of surface wrinkling in various terrestrial and marine organisms to review important morphological parameters and classify functional aspects of surface wrinkles in relation to the size and ecology of organisms. Building on our previous and current experimental studies, we explore case studies on nano/micro-scale wrinkles in biofilms, plant surfaces, and basking shark filter structures to compare developmental and structure-vs-function aspects of wrinkles with vastly different size scales and environmental demands. In doing this and by contrasting wrinkle development in soft and hard biological systems, we provide a template of structure-function relationships of biological surface wrinkles and an outlook for functionalized wrinkled biomimetic surfaces.

Robust and Long-Term Cellular Protein and Enzymatic Activity Preservation in Biomineralized Mammalian Cells.

Preservation of evolved biological structure and function in robust engineering materials is of interest for storage of biological samples before diagnosis and development of vaccines, sensors, and enzymatic reactors and has the potential to avoid cryopreservation and its associated cold-chain issues. Here, we demonstrate that "freezing cells in amorphous silica" is a powerful technique for long-term preservation of whole mammalian cell proteomic structure and function at room temperature. Biomimetic silicification employs the crowded protein microenvironment of mammalian cells as a catalytic framework to proximally transform monomeric silicic acid into silicates forming a nanoscopic silica shell over all biomolecular interfaces. Silicification followed by dehydration preserves and passivates proteomic information within a nanoscale thin silica coating that exhibits size selective permeability (<3.6 nm), preventing protein leaching and protease degradation of cellular contents, while providing access of small molecular constituents for cellular enzymatic reaction. Exposure of dehydrated silicified cells to mild etchant or prolonged hydrolysis removes the silica, completely rerevealing biomolecular components and restoring their accessibility and functionality.

Rapid collagen-directed mineralization of calcium fluoride nanocrystals with periodically patterned nanostructures.

Collagen fibrils present periodic structures, which provide space for intrafibrillar growth of oriented hydroxyapatite nanocrystals in bone and contribute to the good mechanical properties of bone. However, there are not many reports focused on bioprocess-inspired synthesis of non-native inorganic materials inside collagen fibrils and detailed forming processes of crystals inside collagen fibrils remain poorly understood. Herein, the rapid intrafibrillar mineralization of calcium fluoride nanocrystals with a periodically patterned nanostructure is demonstrated. The negatively charged calcium fluoride precursor phase infiltrates collagen fibrils through the gap zones creating an intricate periodic mineralization pattern. Later, the nanocrystals initially filling the gap zones only expand gradually into the remaining space within the collagen fibrils. Mineralized tendons with organized calcium fluoride nanocrystals acquire mechanical properties (indentation elastic modulus ∼25.1 GPa and hardness ∼1.5 GPa) comparable or even superior to those of native human dentin and lamellar bone. Understanding the mineral growth processes in collagen may facilitate the development of tissue engineering and repairing.

Implementing Zn2+ ion and pH-value control into artificial mussel glue proteins by abstracting a His-rich domain from preCollagen.

A His-rich domain of preCollagen-D found in byssal threads is derivatized with Cys and Dopa flanks to allow for mussel-inspired polymerization. Artificial mussel glue proteins are accessed that combine cysteinyldopa for adhesion with sequences for pH or Zn2+ induced ß-sheet formation. The artificial constructs show strong adsorption to Al2O3, the resulting coatings tolerate hypersaline conditions and cohesion is improved by activating the ß-sheet formation, that enhances E-modulus up to 60%.

Molding and Encoding Carbon Nitride-Containing Edible Oil Liquid Objects via Interfacial Toughening in Waterborne Systems.

Charge interaction-driven jamming of nanoparticle monolayers at the oil-water interface can be employed as a method to mold liquids into tailored stable 3D liquid objects. Here, 3D liquid objects are fabricated via a combination of biocompatible aqueous poly(vinyl sulfonic acid, sodium salt) solution and a colloidal dispersion of highly fluorescent organo-modified graphitic carbon nitride (g-C3N4) in edible sunflower oil. The as-formed liquid object shows stability in a broad pH range, as well as flexible pathways for efficient exchange of molecules at the liquid-liquid interphase, which allows for photodegradation of rhodamine B at the interface via visible light irradiation that also enables an encoding concept. The g-C3N4-based liquid objects point toward various applications, for example, all-liquid biphasic photocatalysis, artificial compartmentalized systems, liquid-liquid printing, or bioprinting.

Shape-preserving erosion controlled by the graded microarchitecture of shark tooth enameloid.

The teeth of all vertebrates predominantly comprise the same materials, but their lifespans vary widely: in stark contrast to mammals, shark teeth are functional only for weeks, rather than decades, making lifelong durability largely irrelevant. However, their diets are diverse and often mechanically demanding, and as such, their teeth should maintain a functional morphology, even in the face of extremely high and potentially damaging contact stresses. Here, we reconcile the dilemma between the need for an operative tooth geometry and the unavoidable damage inherent to feeding on hard foods, demonstrating that the tooth cusps of Port Jackson sharks, hard-shelled prey specialists, possess unusual microarchitecture that controls tooth erosion in a way that maintains functional cusp shape. The graded architecture in the enameloid provokes a location-specific damage response, combining chipping of outer enameloid and smooth wear of inner enameloid to preserve an efficient shape for grasping hard prey. Our discovery provides experimental support for the dominant theory that multi-layered tooth enameloid facilitated evolutionary diversification of shark ecologies.

Endoskeletal mineralization in chimaera and a comparative guide to tessellated cartilage in chondrichthyan fishes (sharks, rays and chimaera).

An accepted uniting character of modern cartilaginous fishes (sharks, rays, chimaera) is the presence of a mineralized, skeletal crust, tiled by numerous minute plates called tesserae. Tesserae have, however, never been demonstrated in modern chimaera and it is debated whether the skeleton mineralizes at all. We show for the first time that tessellated cartilage was not lost in chimaera, as has been previously postulated, and is in many ways similar to that of sharks and rays. Tesserae in Chimaera monstrosa are less regular in shape and size in comparison to the general scheme of polygonal tesserae in sharks and rays, yet share several features with them. For example, Chimaera tesserae, like those of elasmobranchs, possess both intertesseral joints (unmineralized regions, where fibrous tissue links adjacent tesserae) and recurring patterns of local mineral density variation (e.g. Liesegang lines, hypermineralized 'spokes'), reflecting periodic accretion of mineral at tesseral edges as tesserae grow. Chimaera monstrosa's tesserae, however, appear to lack the internal cell networks that characterize tesserae in elasmobranchs, indicating fundamental differences among chondrichthyan groups in how calcification is controlled. By compiling and comparing recent ultrastructure data on tesserae, we also provide a synthesized, up-to-date and comparative glossary on tessellated cartilage, as well as a perspective on the current state of research into the topic, offering benchmark context for future research into modern and extinct vertebrate skeletal tissues.

Toward Artificial Mussel-Glue Proteins: Differentiating Sequence Modules for Adhesion and Switchable Cohesion.

Artificial mussel-glue proteins with pH-triggered cohesion control were synthesized by extending the tyrosinase activated polymerization of peptides to sequences with specific modules for cohesion control. The high propensity of these sequence sections to adopt ß-sheets is suppressed by switch defects. This allows enzymatic activation and polymerization to proceed undisturbed. The ß-sheet formation is regained after polymerization by changing the pH from 5.5 to 6.8, thereby triggering O→N acyl transfer rearrangements that activate the cohesion mechanism. The resulting artificial mussel glue proteins exhibit rapid adsorption on alumina surfaces. The coatings resist harsh hypersaline conditions, and reach remarkable adhesive energies of 2.64 mJ m-2 on silica at pH 6.8. In in situ switch experiments, the minor pH change increases the adhesive properties of a coating by 300 % and nanoindentation confirms the cohesion mechanism to improve bulk stiffness by around 200 %.

Thermal-Disrupting Interface Mitigates Intercellular Cohesion Loss for Accurate Topical Antibacterial Therapy.

Bacterial infections remain a leading threat to global health because of the misuse of antibiotics and the rise in drug-resistant pathogens. Although several strategies such as photothermal therapy and magneto-thermal therapy can suppress bacterial infections, excessive heat often damages host cells and lengthens the healing time. Here, a localized thermal managing strategy, thermal-disrupting interface induced mitigation (TRIM), is reported, to minimize intercellular cohesion loss for accurate antibacterial therapy. The TRIM dressing film is composed of alternative microscale arrangement of heat-responsive hydrogel regions and mechanical support regions, which enables the surface microtopography to have a significant effect on disrupting bacterial colonization upon infrared irradiation. The regulation of the interfacial contact to the attached skin confines the produced heat and minimizes the risk of skin damage during thermoablation. Quantitative mechanobiology studies demonstrate the TRIM dressing film with a critical dimension for surface features plays a critical role in maintaining intercellular cohesion of the epidermis during photothermal therapy. Finally, endowing wound dressing with the TRIM effect via in vivo studies in S. aureus infected mice demonstrates a promising strategy for mitigating the side effects of photothermal therapy against a wide spectrum of bacterial infections, promoting future biointerface design for antibacterial therapy.

Green biolubricant infused slippery surfaces to combat marine biofouling.

HYPOTHESIS: Marine biofouling is a global, longstanding problem for maritime industries and coastal areas arising from the attachment of fouling organisms onto solid immersed surfaces. Slippery Liquid Infused Porous Surfaces (SLIPS) have recently shown promising capacity to combat marine biofouling. In most SLIPS coatings, the lubricant is a silicone/fluorinated-based synthetic component that may not be fully compatible with the marine life. We hypothesized that eco-friendly biolubricants could be used to replace synthetic lubricants in SLIPS for marine anti-fouling. EXPERIMENTS: We developed SLIPS coatings using oleic acid (OA) and methyl oleate (MO) as infusing phases. The infusion efficiency was verified with confocal microscopy, surface spectroscopy, wetting efficiency, and nanocontact mechanics. Using green mussels as a model organism, we tested the anti-fouling performance of the biolubricant infused SLIPS and verified its non-cytotoxicity against fish gill cells. FINDINGS: We find that UV-treated PDMS infused with MO gives the most uniform infused film, in agreement with the lowest interfacial energy among all surface/biolubricants produced. These surfaces exhibit efficient anti-fouling properties, as defined by the lowest number of mussel adhesive threads attached to the surface as well as by the smallest surface/thread adhesion strength. We find a direct correlation between anti-fouling performance and the substrate/biolubricant interfacial energy.

SupraCells: Living Mammalian Cells Protected within Functional Modular Nanoparticle-Based Exoskeletons.

Creating a synthetic exoskeleton from abiotic materials to protect delicate mammalian cells and impart them with new functionalities could revolutionize fields like cell-based sensing and create diverse new cellular phenotypes. Herein, the concept of "SupraCells," which are living mammalian cells encapsulated and protected within functional modular nanoparticle-based exoskeletons, is introduced. Exoskeletons are generated within seconds through immediate interparticle and cell/particle complexation that abolishes the macropinocytotic and endocytotic nanoparticle internalization pathways that occur without complexation. SupraCell formation is shown to be generalizable to wide classes of nanoparticles and various types of cells. It induces a spore-like state, wherein cells do not replicate or spread on surfaces but are endowed with extremophile properties, for example, resistance to osmotic stress, reactive oxygen species, pH, and UV exposure, along with abiotic properties like magnetism, conductivity, and multifluorescence. Upon decomplexation cells return to their normal replicative states. SupraCells represent a new class of living hybrid materials with a broad range of functionalities.

A diecast mineralization process forms the tough mantis shrimp dactyl club.

Biomineralization, the process by which mineralized tissues grow and harden via biogenic mineral deposition, is a relatively lengthy process in many mineral-producing organisms, resulting in challenges to study the growth and biomineralization of complex hard mineralized tissues. Arthropods are ideal model organisms to study biomineralization because they regularly molt their exoskeletons and grow new ones in a relatively fast timescale, providing opportunities to track mineralization of entire tissues. Here, we monitored the biomineralization of the mantis shrimp dactyl club-a model bioapatite-based mineralized structure with exceptional mechanical properties-immediately after ecdysis until the formation of the fully functional club and unveil an unusual development mechanism. A flexible membrane initially folded within the club cavity expands to form the new club's envelope. Mineralization proceeds inwards by mineral deposition from this membrane, which contains proteins regulating mineralization. Building a transcriptome of the club tissue and probing it with proteomic data, we identified and sequenced Club Mineralization Protein 1 (CMP-1), an abundant mildly phosphorylated protein from the flexible membrane suggested to be involved in calcium phosphate mineralization of the club, as indicated by in vitro studies using recombinant CMP-1. This work provides a comprehensive picture of the development of a complex hard tissue, from the secretion of its organic macromolecular template to the formation of the fully functional club.

Biological composites-complex structures for functional diversity.

The bulk of Earth's biological materials consist of few base substances-essentially proteins, polysaccharides, and minerals-that assemble into large varieties of structures. Multifunctionality arises naturally from this structural complexity: An example is the combination of rigidity and flexibility in protein-based teeth of the squid sucker ring. Other examples are time-delayed actuation in plant seed pods triggered by environmental signals, such as fire and water, and surface nanostructures that combine light manipulation with mechanical protection or water repellency. Bioinspired engineering transfers some of these structural principles into technically more relevant base materials to obtain new, often unexpected combinations of material properties. Less appreciated is the huge potential of using bioinspired structural complexity to avoid unnecessary chemical diversity, enabling easier recycling and, thus, a more sustainable materials economy.

Biomechanical Design of the Mantis Shrimp Saddle: A Biomineralized Spring Used for Rapid Raptorial Strikes.

Stomatopods deliver one of the fastest strikes in the animal kingdom using their powerful "dactyl clubs." This kinematic performance is enabled by a power amplification device whereby elastic energy is stored in a saddle-shape mineralized bilayer structure. We combined a set of comprehensive micro-mechanical measurements with finite element modeling (FEM) to quantitatively elucidate the saddle biomechanical design. Dynamic nano-scale testing reveals that viscoelastic dissipation is minimized in the highly mineralized layer, whereas micro-bending experiments on miniature cantilevers highlight the critical role of the bilayer arrangement in optimizing storage of elastic energy. FEM shows that the saddle shape prevents stress concentration and the stresses remain well within the elastic range during loading, while the neutral surface coincides with the bilayer interface to prevent interfacial delamination. The study unveils the multi-scale design behind the intriguing ability of the saddle to store a high density of elastic energy using stiff but intrinsically brittle materials. VIDEO ABSTRACT.

Multi-scale structural design and biomechanics of the pistol shrimp snapper claw.

The Arthropoda, the largest phylum of the Animal Kingdom, have successfully evolved to survive various ecological constraints under a wide range of environmental conditions. Central to this survival are the structural designs developed in their exoskeletons and their raptorial appendages for protection and hunting. One such example, the pistol shrimp, is a shallow-water crustacean that is well-known for its aggressive hunting behavior, using its snapper claw to trigger the nucleation of cavitation bubbles that strike targets. In this study, we conducted a multi-scale structural/nanomechanics relationship study of this biotool to analyze its mechanical response to contact stresses. We found that the pistol shrimp snapper claw, which exhibits the capacity to emit a high-velocity water jet during rapid closure actions, is more brittle than other mineralized biotools, exhibiting accelerated wear damage under contact stresses. However, due to an angular offset between the dactylus and pollex of the snapper claw, the appendage never engages in any mechanical contact during the snapping action. This feature is in stark contrast to that reported in other fast raptorial appendages of crustaceans, notably the mantis shrimp dactyl club, which is designed to shatter close range targets in contact mode and exhibits a superior resistance to contact damage and wear. These findings suggest that adaptation of hunting appendages goes beyond their macroscopic morphology, and that multi-scale structural design concomitantly adapted to function, with enhanced structural complexification for tools that are subjected to more intense contact stresses. STATEMENT OF SIGNIFICANCE: The evolution success of crustaceans is largely due to the diversification of their mineralized exoskeletons and hunting appendages, which exhibit a large palette of morphometric characteristics that have adapted to meet particular functions. We explored the "snapper claw" of the pistol shrimp, which is used to generate cavitation bubbles and strike prey. Our multi-scale structure-property relationship study reveals that the snapper claw is more brittle than other fast raptorial appendages - such as the stomatopod dactyl club - because it is not directly subjected to direct contact forces during action. This study implies that when higher mechanical stresses are needed to meet the function, the internal structure is built of a more complex architecture that allows to mitigate internal structural damage.

Squid Suckerin Biomimetic Peptides Form Amyloid-like Crystals with Robust Mechanical Properties.

We present the self-assembly of fibers formed from a peptide sequence (A1H1) derived from suckerin proteins of squid sucker ring teeth (SRT). SRT are protein-only biopolymers with an unconventional set of physicochemical and mechanical properties including high elastic modulus coupled with thermoplastic behavior. We have identified a conserved peptide building block from suckerins that possess the ability to assemble into materials with similar mechanical properties as the native SRT. A1H1 displays amphiphilic characteristics and self-assembles from the bottom-up into mm-scale fibers initiated by the addition of a polar aprotic solvent. A1H1 fibers are thermally resistant up to 239 °C, coupled with an elastic modulus of ∼7.7 GPa, which can be explained by the tight packing of ß-sheet-enriched crystalline building blocks as identified by wide-angle X-ray scattering (WAXS), with intersheet and interstrand distances of 5.37 and 4.38 Å, respectively. A compact packing of the peptides at their Ala-rich terminals within the fibers was confirmed from molecular dynamics simulations, and we propose a hierarchical model of fiber assembly of the mature peptide fiber.