Shahrouz Amini

Accelerating the design of biomimetic materials by integrating RNA-seq with proteomics and materials science

Efforts to engineer new materials inspired by biological structures are hampered by the lack of genomic data from many model organisms studied in biomimetic research. Here we show that biomimetic engineering can be accelerated by integrating high-throughput RNA-seq with proteomics and advanced materials characterization. This approach can be applied to a broad range of systems, as we illustrate by investigating diverse high-performance biological materials involved in embryo protection, adhesion and predation. In one example, we rapidly engineer recombinant squid sucker ring teeth proteins into a range of structural and functional materials, including nanopatterned surfaces and photo-cross-linked films that exceed the mechanical properties of most natural and synthetic polymers. Integrating RNA-seq with proteomics and materials science facilitates the molecular characterization of natural materials and the effective translation of their molecular designs into a wide range of bio-inspired materials.

Bio-Inspired Mechanotactic Hybrids for Orchestrating Traction-Mediated Epithelial Migration.

A platform of mechanotactic hybrids is established by projecting lateral gradients of apparent interfacial stiffness onto the planar surface of a compliant hydrogel layer using an underlying rigid substrate with microstructures inherited from 3D printed molds. Using this platform, the mechanistic coupling of epithelial migration with the stiffness of the extracellular matrix (ECM) is found to be independent of the interfacial compositional and topographical cues.

Textured fluorapatite bonded to calcium sulphate strengthen stomatopod raptorial appendages

Stomatopods are shallow-water crustaceans that employ powerful dactyl appendages to hunt their prey. Deployed at high velocities, these hammer-like clubs or spear-like devices are able to inflict substantial impact forces. Here we demonstrate that dactyl impact surfaces consist of a finely-tuned mineral gradient, with fluorapatite substituting amorphous apatite towards the outer surface. Raman spectroscopy measurements show that calcium sulphate, previously not reported in mechanically active biotools, is co-localized with fluorapatite. Ab initio computations suggest that fluorapatite/calcium sulphate interfaces provide binding stability and promote the disordered-to-ordered transition of fluorapatite. Nanomechanical measurements show that fluorapatite crystalline orientation correlates with an anisotropic stiffness response and indicate significant differences in the fracture tolerance between the two types of appendages. Our findings shed new light on the crystallochemical and microstructural strategies allowing these intriguing biotools to optimize impact forces, providing physicochemical information that could be translated towards the synthesis of impact-resistant functional materials and coatings.

Wear and abrasion resistance selection maps of biological materials

The mechanical design of biological materials has generated widespread interest in recent years, providing many insights into their intriguing structure-property relationships. A critical characteristic of load-bearing materials, which is central to the survival of many species, is their wear and abrasion tolerance. In order to be fully functional, protective armors, dentitious structures and dynamic appendages must be able to tolerate repetitive contact loads without significant loss of materials or internal damage. However, very little is known about this tribological performance. Using a contact mechanics framework, we have constructed materials selection charts that provide general predictions about the wear performance of biological materials as a function of their fundamental mechanical properties. One key assumption in constructing these selection charts is that abrasion tolerance is governed by the first irreversible damage at the contact point. The maps were generated using comprehensive data from the literature and encompass a wide range of materials, from heavily mineralized to fully organic materials. Our analysis shows that the tolerance of biological materials against abrasion depends on contact geometry, which is ultimately correlated to environmental and selective pressures. Comparisons with experimental data from nanoindentation experiments are also drawn in order to verify our predictions. With the increasing amount of data available for biological materials also comes the challenge of selecting relevant model systems for bioinspired materials engineering. We suggest that these maps will be able to guide this selection by providing an overview of biological materials that are predicted to exhibit the best abrasion tolerance, which is of fundamental interest for a wide range of applications, for instance in restorative implants and protective devices.

The role of quasi-plasticity in the extreme contact damage tolerance of the stomatopod dactyl club

The structure of the stomatopod dactyl club--an ultrafast, hammer-like device used by the animal to shatter hard seashells--offers inspiration for impact-tolerant ceramics. Here, we present the micromechanical principles and related micromechanisms of deformation that impart the club with high impact tolerance. By using depth-sensing nanoindentation with spherical and sharp contact tips in combination with post-indentation residual stress mapping by Raman microspectroscopy, we show that the impact surface region of the dactyl club exhibits a quasi-plastic contact response associated with the interfacial sliding and rotation of fluorapatite nanorods, endowing the club with localized yielding. We also show that the subsurface layers exhibit strain hardening by microchannel densification, which provides additional dissipation of impact energy. Our findings suggest that the clubs macroscopic size is below the critical size above which Hertzian brittle cracks are nucleated.

Structural, Nanomechanical, and Computational Characterization of d , l -Cyclic Peptide Assemblies

The rigid geometry and tunable chemistry of D,L-cyclic peptides makes them an intriguing building-block for the rational design of nano- and microscale hierarchically structured materials. Herein, we utilize a combination of electron microscopy, nanomechanical characterization including depth sensing-based bending experiments, and molecular modeling methods to obtain the structural and mechanical characteristics of cyclo-[(Gln-D-Leu)4] (QL4) assemblies. QL4 monomers assemble to form large, rod-like structures with diameters up to 2 μm and lengths of tens to hundreds of micrometers. Image analysis suggests that large assemblies are hierarchically organized from individual tubes that undergo bundling to form larger structures. With an elastic modulus of 11.3 ± 3.3 GPa, hardness of 387 ± 136 MPa and strength (bending) of 98 ± 19 MPa the peptide crystals are among the most robust known proteinaceous micro- and nanofibers. The measured bending modulus of micron-scale fibrils (10.5 ± 0.9 GPa) is in the same range as the Youngs modulus measured by nanoindentation indicating that the robust nanoscale network from which the assembly derives its properties is preserved at larger length-scales. Materials selection charts are used to demonstrate the particularly robust properties of QL4 including its specific flexural modulus in which it outperforms a number of biological proteinaceous and nonproteinaceous materials including collagen and enamel. The facile synthesis, high modulus, and low density of QL4 fibers indicate that they may find utility as a filler material in a variety of high efficiency, biocompatible composite materials.

Multi-scale thermal stability of a hard thermoplastic protein-based material

Although thermoplastic materials are mostly derived from petro-chemicals, it would be highly desirable, from a sustainability perspective, to produce them instead from renewable biopolymers. Unfortunately, biopolymers exhibiting thermoplastic behaviour and which preserve their mechanical properties post processing are essentially non-existent. The robust sucker ring teeth (SRT) from squid and cuttlefish are one notable exception of thermoplastic biopolymers. Here we describe thermoplastic processing of squid SRT via hot extrusion of fibres, demonstrating the potential suitability of these materials for large-scale thermal forming. Using high-resolution in situ X-ray diffraction and vibrational spectroscopy, we elucidate the molecular and nanoscale features responsible for this behaviour and show that SRT consist of semi-crystalline polymers, whereby heat-resistant, nanocrystalline β-sheets embedded within an amorphous matrix are organized into a hexagonally packed nanofibrillar lattice. This study provides key insights for the molecular design of biomimetic protein- and peptide-based thermoplastic structural biopolymers with potential biomedical and 3D printing applications.

Orientational Coupling Locally Orchestrates a Cell Migration Pattern for Re-Epithelialization

Re-epithelialization by collective migration of epithelial cells over a heterogeneous environment to restore tissue integrity and functions is critical for development and regeneration. Here, it is reported that the spatial organization of adjacent adherent paths within sparsely distributed extracellular matrix (ECM) has a significant impact on the orientational coupling between cell polarization and collective cell migration. This coupling effect determines the migration pattern for human keratinocytes to regain their cohesion, which impacts the occupancy of epithelial bridge and the migration velocity in wound repair. Statistical studies suggest the converging organization of ECM, in which adjacent paths become closer to each other and finally converge to a junctional point, facilitating collective cell migration mostly within variable ECM organization, as the polarization of the advancing cell sheet is remodeled to align along the direction of cell migration. The findings may help to design implantable ECM to optimize efficient skin regeneration.

Fabrication of a 3D hair follicle‐like hydrogel by soft lithography

Hair follicle transplantation is often used in the treatment of androgenetic alopecia (AGA). However, the only source of hair follicles is from human donors themselves, which limits the application of this approach. One possible solution is to reconstitute hair follicle from dissociated cells. Currently, a number of microscale technologies have been developed to create size and shape controlled microenvironments in tissue engineering. Photopolymerizable PEGDA hydrogels are often selected as promising scaffolds in engineered microtissues due to their biocompatibility and adjustable mechanical properties. Here, we fabricated an array of PEGDA microwells with center islets that mimic the architecture of human hair follicles using soft lithography. Dermal and epithelial cells were seeded in different compartments of the microstructured mould to mimic mesenchymal and epithelial compartmentalization in native hair follicles. We demonstrated that these compartmentalized microstructures support cell proliferation and cell survival over 14 days, and spreading of dermal fibroblasts was observed. This hydrogel micromould provides a potentially useful tool for engineering 3D hair follicle-mimicking complex cultures in vitro.

Squid suckerin microneedle arrays for tunable drug release

Microneedles are increasingly used in transdermal delivery of therapeutic agents due to the elimination of first-pass metabolism, simplicity of operation, and lack of pain, which collectively lead to improved patient compliance. However, microneedles are still met by challenges with regard to the choice of biocompatible materials and the control of drug release profiles. Herein, we tackle these limitations by producing microneedles from a biocompatible robust biopolymer, namely squid sucker ring teeth (SRT) proteins (suckerins), using a soft lithography method. Taking advantage of the modular sequence design of suckerins leading to their self-assembly into β-sheet enriched structures, suckerin microneedles display an accurate replication of their templates with robust mechanical properties, endowing them with a high skin penetration capability. Critically, the β-sheet content in the microneedles can be modulated by varying the solvent conditions, which allows tuning of the mechanical response, and in turn the drug release rates by more than one order of magnitude. In vitro skin permeation studies of suckerin microneedles using human cadaver skin samples suggest a fast onset and enhanced skin permeation of drugs compared to flat patches. The skin permeation can also be tailored 10-fold by applying hydrogen bond disruptor solutions. As a proof-of-concept, the anti-bacterial drug kanamycin is encapsulated within the microneedles, leading to efficient anti-bacterial activity and offering an additional benefit to further minimize the risk of infections caused by microneedle-based drug delivery systems. Lastly, suckerin microneedles are found to be biocompatible in cell culture studies, opening the door to further clinical applications.