In this study, the pH-dependent friction of layer-by-layer assemblies of poly(allylamine hydrochloride) and poly(acrylic acid) (PAH/PAA) are quantified for microtube array structures via experimental and simulated lateral force microscopy (LFM). A novel coarse-grain tube model is developed, utilizing a molecular dynamics (MD) framework with a Hertzian soft contact potential (such that F ∼ δ3/2) to allow the efficient dynamic simulation of 3D arrays consisting of hundreds of tubes at micrometer length scales. By quantitatively comparing experimental LFM and computational results, the coupling between geometry (tube spacing and swelling) and material properties (intrinsic stiffness) results in a transition from bending dominated deformation to bending combined with inter-tube contact, independent of material adhesion assumptions. Variation of tube spacing (and thus control of contact) can be used to exploit the normal and lateral resistance of the tube arrays as a function of pH (2.0/5.5), beyond the effect of areal tube density, with increased resistances (potential mutability) up to a factor of ∼60. This study provides a novel modeling platform to assess and design dynamic polyelectrolyte-based substrates/coatings with tailorable stimulus-responsive surface friction. Our results show that micro-geometry can be used alongside stimulus-responsive material changes to amplify and systematically tune mutability.

High resolution surface textures, when rationally designed, provide an attractive surface engineering approach to enhance surface functionalities. Designing smart surfaces by coupling surface texture with shape memory polymers has garnered attention in achieving tunable mechanical properties. We investigate the structure–mechanical property relationships for programmable, shape-memorizing submicron-scale pillar arrays subjected to flat-punch compression. The geometrically-dependent deformation of structured surfaces with two different aspect ratios (250 nm-pillars 1 : 1 and 550 nm-pillars 2.4 : 1) were investigated, and their moduli were found to be lower than that of non-patterned surface. From finite element analysis, the pillar deformation is correlated to a mechanistic transition from a discrete, unidirectional compression of 250 nm-pillars to lateral constraints caused by interpillar contact in 550 nm-pillars. This lateral pillar–pillar contact in the 550 nm-pillars resulted in an increased and maximum strain-dependent modulus but lower elastic recovery and energy dissipation as compared with the 250 nm-pillars. Furthermore, the compressive responses of temporarily shaped pillars (programmed by stretching) were compared with the permanently shaped pillars. The extent of lateral constraints controlled by pillar shape and spacing in 550 nm-pillars was responsible for the modulus differences between the original and stretched patterns, whereas the modulus of 250 nm-pillars remained as a constant value with different levels of stretching. This study provides mechanistic insights into how the mechanical behavior can be modulated by designing the aspect ratio of shape memory pillar arrays and by programming the surface geometry, thus revealing the potential of developing ingenious designs of responsive surfaces sensitive to mechanical deformation.

Poroelastic interactions between interstitial fluid and the extracellular matrix of connective tissues are critical to biological and pathophysiological functions involving solute transport, energy dissipation, self-stiffening and lubrication. However, the molecular origins of poroelasticity at the nanoscale are largely unknown. Here, the broad-spectrum dynamic nanomechanical behavior of cartilage aggrecan monolayer is revealed for the first time, including the equilibrium and instantaneous moduli and the peak in the phase angle of the complex modulus. By performing a length scale study and comparing the experimental results to theoretical predictions, we confirm that the mechanism underlying the observed dynamic nanomechanics is due to solid–fluid interactions (poroelasticity) at the molecular scale. Utilizing finite element modeling, the molecular-scale hydraulic permeability of the aggrecan assembly was quantified (kaggrecan = (4.8 ± 2.8) × 10–15 m4/N·s) and found to be similar to the nanoscale hydraulic permeability of intact normal cartilage tissue but much lower than that of early diseased tissue. The mechanisms underlying aggrecan poroelasticity were further investigated by altering electrostatic interactions between the molecule’s constituent glycosaminoglycan chains: electrostatic interactions dominated steric interactions in governing molecular behavior. While the hydraulic permeability of aggrecan layers does not change across species and age, aggrecan from adult human cartilage is stiffer than the aggrecan from newborn human tissue.Keywords: AFM; aggrecan; cartilage; dynamic loading; hydraulic permeability; nanomechanics; poroelasticity;

Inspired by the overlapping scales found on teleost fish, a new composite architecture explores the mechanics of materials to accommodate both flexibility and protection. These biological structures consist of overlapping mineralized plates embedded in a compliant tissue to form a natural flexible armor which protects underlying soft tissue and vital organs. Here, the functional performance of such armors is investigated, in which the composition, spatial arrangement, and morphometry of the scales provide locally tailored functionality. Fabricated macroscale prototypes and finite element based micromechanical models are employed to measure mechanical response to blunt and penetrating indentation loading. Deformation mechanisms of scale bending, scale rotation, tissue shear, and tissue constraint were found to govern the ability of the composite to protect the underlying substrate. These deformation mechanisms, the resistance to deformation, and the resulting work of deformation can all be tailored by structural parameters including architectural arrangement (angle of the scales, degree of scale overlap), composition (volume fraction of the scales), morphometry (aspect ratio of the scales), and material properties (tissue modulus and scale modulus). In addition, this network of armor serves to distribute the load of a predatory attack over a large area to mitigate stress concentrations. Mechanical characterization of such layered, segmented structures is fundamental to developing design principles for engineered protective systems and composites.

A common feature of the outer layer of protective biological exoskeletons is structural anisotropy. Here, we directly quantify the mechanical anisotropy and fracture of an individual material layer of a hydroxyapatite-based nanocomposite exoskeleton, the outmost ganoine of Polypterus senegalus scale. Uniaxial compression was conducted on cylindrical micropillars of ganoine fabricated via focused ion beam at different orientations relative to the hydroxyapatite rod long axis (θ = 0°, 45°, 90°). Engineering stress versus strain curves revealed significant elastic and plastic anisotropy, off-axial strain hardening, and noncatastrophic crack propagation within ganoine. Off-axial compression (θ = 45°) showed the lowest elastic modulus, E (36.2 ± 1.6 GPa, n ≥ 10, mean ± SEM), and yield stress, σY (0.81 ± 0.02 GPa), while compression at θ = 0° showed the highest E (51.8 ± 1.7 GPa) and σY (1.08 ± 0.05 GPa). A 3D elastic–plastic composite nanostructural finite element model revealed this anisotropy was correlated to the alignment of the HAP rods and could facilitate energy dissipation and damage localization, thus preventing catastrophic failure upon penetration attacks.

It has been hypothesized that predatory threats are a critical factor in the protective functional design of biological exoskeletons or “natural armor”, having arisen through evolutionary processes. Here, the mechanical interaction between the ganoid armor of the predatory fish Polypterus senegalus and one of its current most aggressive threats, a toothed biting attack by a member of its own species (conspecific), is simulated and studied. Finite element analysis models of the quad-layered mineralized scale and representative teeth are constructed and virtual penetrating biting events simulated. Parametric studies reveal the effects of tooth geometry, microstructure and mechanical properties on its ability to effectively penetrate into the scale or to be defeated by the scale, in particular the deformation of the tooth versus that of the scale during a biting attack. Simultaneously, the role of the microstructure of the scale in defeating threats as well as providing avenues of energy dissipation to withstand biting attacks is identified. Microstructural length scale and material property length scale matching between the threat and armor is observed. Based on these results, a summary of advantageous and disadvantageous design strategies for the offensive threat and defensive protection is formulated. Studies of predator-prey threat-protection interactions may lead to insights into adaptive phenotypic plasticity of the tooth and scale microstructure and geometry, “adaptive stalemates” and the so-called evolutionary “arms race”.

Biological exoskeletons, in particular those with unusually robust and multifunctional properties, hold enormous potential for the development of improved load-bearing and protective engineering materials. Here, we report new materials and mechanical design principles of the iron-plated multilayered structure of the natural armor of Crysomallon squamiferum, a recently discovered gastropod mollusc from the Kairei Indian hydrothermal vent field, which is unlike any other known natural or synthetic engineered armor. We have determined through nanoscale experiments and computational simulations of a predatory attack that the specific combination of different materials, microstructures, interfacial geometries, gradation, and layering are advantageous for penetration resistance, energy dissipation, mitigation of fracture and crack arrest, reduction of back deflections, and resistance to bending and tensile loads. The structure-property-performance relationships described are expected to be of technological interest for a variety of civilian and defense applications.

Here we report the formation of stimulus-responsive chemically end-grafted “brush-brushes” by synthesizing, mono thiol(end)-functionalized poly(methacrylic acid-g-ethylene glycol) or poly(MAA-g-EG) comb-type graft copolymers via a combination of protecting group chemistry and atom transfer radical polymerization using the initiator 2-(2,4-dinitrophenylthio)ethyl 2-bromo-2-methyl propionate. The polymers were synthesized with three different molecular weights (15 k, 17 k and 27 k), PEG side chain graft densities (EG/MAA mole ratio = 2.2, 0.4 and 1.9, respectively), and a PEG molecular weight = 1100 and then chemically end-grafted to gold substrates via chemisorption, resulting in molecular separation distances of ∼3–4 nm. pH-Dependent swelling was confirmed to take place gradually above pH 4–5 and quantified by heights measured by contact mode AFM imaging of microcontact printed (µCP) samples. Swelling factors (maximum height/minimum height) were fairly large (3.6–7.3) and a decrease in molecular weight by ∼2× and side chain graft density by ∼4× resulted in a decrease in swelling factor by ∼2×. Layer height versus normal force for all three polymers measured by contact mode atomic force microscope imaging on µCP samples at pH 9 showed a nonlinearly decreasing relationship and complete compression ∼<2 nm for forces >10 nN. At pH 4, all polymer layers were largely collapsed (heights ∼<4 nm) and incompressible (i.e., heights were independent of normal force).

The normal intersurface forces between nanosized probe tips functionalized with COO−- and NH3+-terminated alkanethiol self-assembling monolayers and dense polycrystalline phase pure synthetic hydroxyapatite (HA) were measured via a powerful nanomechanical technique called chemically specific high-resolution force spectroscopy. The data taken on approach of the probe tip to the HA surface was compared to the nonlinear Poisson–Boltzmann-based electrostatic double layer theory to predict the surface charge per unit area of the HA, σHA (C/m2), as a function of ionic strength, position within a variety of grains, and across grain boundaries. The average σHA was found to be ∼−0.02 C/m2 and to vary from −0.0037 to −0.072 C/m2 with nanoscale position in relation to grain boundaries and crystal planes up to −0.19 C/m2/μm. Positional measurement of nanoscale surface properties holds great promise in elucidating the molecular origins of physicochemical processes occurring at the biomaterial interface.

In this study, a quantitative investigation of the microstructure and composition of field-caught marine Gasterosteus aculeatus (threespine stickleback) armor is presented, which provides useful phylogenetic information and insights into biomechanical function. Micro-computed tomography (μCT) was employed to create full three-dimensional images of the dorsal spines and basal plate, lateral plates, pelvic girdle and spines and to assess structural and compositional properties such as the spatial distribution of thickness (∼100–300 μm), the heterogeneous cross-sectional geometry (centrally thickened), plate-to-plate juncture and overlap (∼50% of the plate width), and bone mineral density (634–748 HA/cm3). The convolution of plate geometry in conjunction with plate-to-plate overlap allows a relatively constant armor thickness to be maintained throughout the assembly, promoting spatially homogeneous protection and thereby avoiding weakness at the armor unit interconnections. Plate-to-plate junctures act to register and join the plates while permitting compliance in sliding and rotation in selected directions. Mercury porosimetry was used to determine the pore size distribution and volume percent porosity of the lateral plates (20–35 vol.%) and spines (10–15 vol.%). SEM and μCT revealed a porous, sandwich-like cross-section beneficial for bending stiffness and strength at minimum weight. Back-scattered electron microscopy and energy dispersive X-ray analysis were utilized to quantify the weight percent mineral content (58–68%). Scanning electron microscopy and surface profilometry were used to characterize the interior and exterior surface topography (tubercles) of the lateral plates. The results obtained in this study are discussed in the context of mechanical function, performance, fitness, and survivability.

The nanostructure and nanomechanical properties of aggrecan monomers extracted and purified from human articular cartilage from donors of different ages (newborn, 29 and 38 year old) were directly visualized and quantified via atomic force microscopy (AFM)-based imaging and force spectroscopy. AFM imaging enabled direct comparison of full length monomers at different ages. The higher proportion of aggrecan fragments observed in adult versus newborn populations is consistent with the cumulative proteolysis of aggrecan known to occur in vivo. The decreased dimensions of adult full length aggrecan (including core protein and glycosaminoglycan (GAG) chain trace length, end-to-end distance and extension ratio) reflect altered aggrecan biosynthesis. The demonstrably shorter GAG chains observed in adult full length aggrecan monomers, compared to newborn monomers, also reflects markedly altered biosynthesis with age. Direct visualization of aggrecan subjected to chondroitinase and/or keratanase treatment revealed conformational properties of aggrecan monomers associated with chondroitin sulfate (CS) and keratan sulfate (KS) GAG chains. Furthermore, compressive stiffness of chemically end-attached layers of adult and newborn aggrecan was measured in various ionic strength aqueous solutions. Adult aggrecan was significantly weaker in compression than newborn aggrecan even at the same total GAG density and bath ionic strength, suggesting the importance of both electrostatic and non-electrostatic interactions in nanomechanical stiffness. These results provide molecular-level evidence of the effects of age on the conformational and nanomechanical properties of aggrecan, with direct implications for the effects of aggrecan nanostructure on the age-dependence of cartilage tissue biomechanical and osmotic properties.

Heterogeneity of biological materials, such as bone, tooth, and mollusc shells, plays a key role in determining their mechanical performance (e.g. the strength, damage tolerance, etc.). Here, we quantify heterogeneities in elasticity and inelasticity of bovine cortical bone between 100 nm and a few microns and identify a characteristic length scale (λc) of approximately 200 nm. Below λc the mechanical heterogeneity of bone is pronounced and exhibits a strong nonlinear size-dependence, while above λc the heterogeneity is much less. Such size-dependent heterogeneity benefits the mechanical performance of bone since it not only promotes the energy dissipation at nanoscale, but also suppresses heterogeneity-induced stress concentration and strain localization at larger length scales. This is one of the possible mechanisms functioning at multiple length scales that make bone a well-designed tough natural material. Utilizing experimentally measured data, systematic computational simulations were carried out, showing that the heterogeneity in inelasticity, rather than elasticity, plays a dominant role in promoting energy dissipation during deformation. Possible parameters that determine the inelasticity heterogeneity (e.g. mean value and standard deviation of heterogeneous yield stress) and therefore affect energy dissipation are investigated under typical deformation modes of bone. The analysis presented suggests that there exists an optimum ratio of macroscopic strength to elastic modulus for improving energy dissipation under tension. All these findings are of great value to the design and synthesis of improved bio-inspired composites.

This study investigates the three-dimensional structure of the eight plate exoskeletal (shell) assembly of the chiton Tonicella marmorea. X-ray micro-computed tomography and 3D printing elucidate the mechanism of conformational change from a passive (slightly curved, attached to surface) to a defensive (rolled, detached from surface) state of the plate assembly. The passive and defensive conformations exhibited differences in longitudinal curvature index (0.43 vs. 0.70), average plate-to-plate overlap (∼62% vs. ∼48%), cross-sectional overlap heterogeneity (60–82.5% vs. 0–90%, fourth plate), and plate-to-plate separation distance (100% increase in normalized separation distance between plates 4 and 5), respectively. The plate-to-plate interconnections consist of two rigid plates joined by a compliant, actuating muscle, analogous to a geometrically structured shear lap joint. This work provides an understanding of how T. marmorea achieves the balance between mobility and protection. In the passive state, the morphometry of the plates and plate-to-plate interconnections results in an approximately continuous curvature and constant armor thickness, resulting in limited mobility but maximum protection. In the defensive state, the underlying soft tissues gain protection and the chiton gains mobility through tidal flow, but regions of vulnerability open dorsally, due to the increase in plate-to-plate separation and decrease in plate-to-plate overlap. Lastly, experiments using optical and scanning electron microscopy, mercury porosimetry, and Fourier-transform infrared spectroscopy explore the microstructure and spatial distribution of the six layers within the intermediate plates, the role of multilayering in resisting predatory attacks, and the detection of chitin as a major component of the intra-plate organic matrix and girdle.

Many armored fish scale assemblies use geometric heterogeneity of subunits as a design parameter to provide tailored biomechanical flexibility while maintaining protection from external penetrative threats. This study analyzes the spatially varying shape of individual ganoid scales as a structural element in a biological system, the exoskeleton of the armored fish Polypterus senegalus (bichir). X-ray microcomputed tomography is used to generate digital 3D reconstructions of the mineralized scales. Landmark-based geometric morphometrics is used to measure the geometric variation among scales and to define a set of geometric parameters to describe shape variation. A formalism using continuum mechanical strain analysis is developed to quantify the spatial geometry change of the scales and illustrate the mechanisms of shape morphing between scales. Five scale geometry variants are defined (average, anterior, tail, ventral, and pectoral fin) and their functional implications are discussed in terms of the interscale mobility mechanisms that enable flexibility within the exoskeleton. The results suggest that shape variation in materials design, inspired by structural biological materials, can allow for tunable behavior in flexible composites made of segmented scale assemblies to achieve enhanced user mobility, custom fit, and flexibility around joints for a variety of protective applications.

In this study, atomic force microscopy-based dynamic oscillatory and force-relaxation indentation was employed to quantify the time-dependent nanomechanics of native (untreated) and proteoglycan (PG)-depleted cartilage disks, including indentation modulus Eind, force-relaxation time constant τ, magnitude of dynamic complex modulus |E∗|, phase angle δ between force and indentation depth, storage modulus E′, and loss modulus E″. At ∼2 nm dynamic deformation amplitude, |E∗| increased significantly with frequency from 0.22 ± 0.02 MPa (1 Hz) to 0.77 ± 0.10 MPa (316 Hz), accompanied by an increase in δ (energy dissipation). At this length scale, the energy dissipation mechanisms were deconvoluted: the dynamic frequency dependence was primarily governed by the fluid-flow-induced poroelasticity, whereas the long-time force relaxation reflected flow-independent viscoelasticity. After PG depletion, the change in the frequency response of |E∗| and δ was consistent with an increase in cartilage local hydraulic permeability. Although untreated disks showed only slight dynamic amplitude-dependent behavior, PG-depleted disks showed great amplitude-enhanced energy dissipation, possibly due to additional viscoelastic mechanisms. Hence, in addition to functioning as a primary determinant of cartilage compressive stiffness and hydraulic permeability, the presence of aggrecan minimized the amplitude dependence of |E∗| at nanometer-scale deformation.

Dynamic nanomechanical properties of bovine bone marrow stromal cells (BMSCs) and their newly synthesized cartilage-like matrices were studied at nanometer scale deformation amplitudes. The increase in their dynamic modulus, |E⁎| (e.g., 2.4±0.4 kPa at 1 Hz to 9.7±0.2 kPa at 316 Hz at day 21, mean±SEM), and phase angle, δ, (e.g., 15±2° at 1 Hz to 74±1° at 316 Hz at day 21) with increasing frequency were attributed to the fluid flow induced poroelasticity, governed by both the newly synthesized matrix and the intracellular structures. The absence of culture duration dependence suggested that chondrogenesis of BMSCs had not yet resulted in the formation of a well-organized matrix with a hierarchical structure similar to cartilage. BMSC-matrix composites demonstrated different poro-viscoelastic frequency-dependent mechanical behavior and energy dissipation compared to chondrocyte-matrix composites due to differences in matrix molecular constituents, structure and cell properties. This study provides important insights into the design of optimal protocols for tissue-engineered cartilage products using chondrocytes and BMSCs.

Here it is reported that aggrecan, the highly negatively charged macromolecule in the cartilage extracellular matrix, undergoes Ca2+-mediated self-adhesion after static compression even in the presence of strong electrostatic repulsion in physiological-like solution conditions. Aggrecan was chemically end-attached onto gold-coated planar silicon substrates and gold-coated microspherical atomic force microscope probe tips (end radius R ≈ 2.5 μm) at a density (∼40 mg/mL) that simulates physiological conditions in the tissue (∼20–80 mg/mL). Colloidal force spectroscopy was employed to measure the adhesion between opposing aggrecan monolayers in NaCl (0.001–1.0 M) and NaCl + CaCl2 ([Cl−] = 0.15 M, [Ca2+] = 0 – 75 mM) aqueous electrolyte solutions. Aggrecan self-adhesion was found to increase with increasing surface equilibration time upon compression (0–30 s). Hydrogen bonding and physical entanglements between the chondroitin sulfate-glycosaminoglycan side chains are proposed as important factors contributing to aggrecan self-adhesion. Self-adhesion was found to significantly increase with decreasing bath ionic strength (and hence, electrostatic double-layer repulsion), as well as increasing Ca2+ concentration due to the additional ion-bridging effects. It is hypothesized that aggrecan self-adhesion, and the macromolecular energy dissipation that results from this self-adhesion, could be important factors contributing to the self-assembled architecture and integrity of the cartilage extracellular matrix in vivo.