Biological Composite Materials Research Articles

Structural diversity of crustacean exoskeletons and its implications for biomimetics.

The crustacean cuticle is a biological composite material consisting of chitin-protein fibres in a mineralized matrix. Recent research has revealed a surprising range of fibre architectures and mineral compositions of crustacean skeletal structures adapted to various mechanical demands. It is becoming increasingly clear that the organic fibres in the cuticle may be organized in patterns differing from the standard twisted plywood model. Observed fibre architectures in protruding skeletal structures include longitudinal and circular parallel fibre arrays. Skeletal minerals often include calcium phosphates in addition to calcium carbonates. Furthermore, skeletal properties are affected by protein cross-linking, which replaces mineralization as a stiffening mechanism in some structures. Several common structural motifs, such as the stiffening of the outer skeletal layers, the incorporation of non-mineralized cuticle in exposed structures, and interchanging layers of parallel fibres and the twisted plywood structure, can be identified in skeletal elements with similar functions. These evolutionary solutions have the potential for biomimetic applications, particularly as manufacturing technologies advance. To make use of this potential, we need to understand the processes behind the formation of the crustacean exoskeleton and determine which features are truly adaptive and worth mimicking.

3D-printed nanohydroxyapatite/methylacrylylated silk fibroin scaffold for repairing rat skull defects

The repair of bone defects remains a major challenge in the clinic, and treatment requires bone grafts or bone replacement materials. Existing biomaterials have many limitations and cannot meet the various needs of clinical applications. To treat bone defects, we constructed a nanohydroxyapatite (nHA)/methylacrylylated silk fibroin (MASF) composite biological scaffold using photocurable 3D printing technology. In this study, scanning electron microscopy (SEM) was used to detect the changes in the morphological structure of the composite scaffold with different contents of nanohydroxyapatite, and FTIR was used to detect the functional groups and chemical bonds in the composite scaffold to determine the specific components of the scaffold. In in vitro experiments, bone marrow mesenchymal stem cells from SD rats were cocultured with scaffolds soaking solution, and the cytotoxicity, cell proliferation, Western blot analysis, Quantitative real-time PCR analysis, bone alkaline phosphatase activity and alizarin red staining of scaffolds were detected to determine the biocompatibility of scaffolds and the effect of promoting proliferation and osteogenesis of bone marrow mesenchymal stem cells in vitro. In the in vivo experiment, the skull defect was constructed by adult SD rats, and the scaffold was implanted into the skull defect site. After 4 weeks and 8 weeks of culture, the specific osteogenic effect of the scaffold in the skull defect site was detected by animal micro-CT, hematoxylin and eosin (HE) staining and Masson's staining. Through the analysis of the morphological structure of the scaffold, we found that the frame supported good retention of the lamellar structure of silk fibroin, when mixed with nHA, the surface of the stent was rougher, the cell contact area increased, and cell adhesion and lamellar microstructure for cell migration and proliferation of the microenvironment provided a better space. FTIR results showed that the scaffold completely retained the β -folded structure of silk fibroin, and the scaffold composite was present without obvious impurities. The staining results of live/dead cells showed that the constructed scaffolds had no significant cytotoxicity, and thw CCK-8 assay also showed that the constructed scaffolds had good biocompatibility. The results of osteogenic induction showed that the scaffold had good osteogenic induction ability. Moreover, the results also showed that the scaffold with a MASF: nHA ratio of 1: 0.5 (SFH) showed better osteogenic ability. The micro-CT and bone histometric results were consistent with the in vitro results after stent implantation, and there was more bone formation at the bone defect site in the SFH group.This research used photocurable 3D printing technology to successfully build an osteogenesis bracket. The results show that the constructed nHA/MASF biological composite material, has good biocompatibility and good osteogenesis function. At the same time, in the microenvironment, the material can also promote bone defect repair and can potentially be used as a bone defect filling material for bone regeneration applications.

Effect of HA Content on Microstructure and Properties of Ti-27Nb-17Ta-8Zr/HA Composite.

In this paper, Ti-27Nb-17Ta-8Zr/HA series composite materials were prepared by spark plasma sintering (SPS) technology. The medical titanium alloy (Ti-27Nb-17Ta-8Zr) with good mechanical properties, wear resistance, and corrosion resistance was combined with the hydroxyapatite (HA) bioactive ceramic with high biological activity and bone-binding ability. Moreover, the density, microstructure evolution, metal/ceramic reaction, mechanical behavior, in vitro bioactivity, and influencing mechanisms of composite materials with different HA contents were studied. The research results indicate that all biological composite materials are composed of β-Ti solution, α-Ti, and ceramic phases (Ti2O, CaTiO3, CaO, TixPy). With the increase of HA content, the compressive strength and yield strength of the composite material show a trend of first increasing, then decreasing, and then slowly increasing. After soaking in SBF artificial simulated body fluid for 5 days, the deposition of elements such as Ca and P on the surface significantly increased, while elements such as Ti, Nb, Ta, and Zr were evenly distributed in the matrix, demonstrating good in vitro mineralization ability and facilitating the attachment and growth of osteoblasts.

Cold sintering constructed in situ drug-loaded high strength HA-PLA composites: Potential bone substitution material

Hydroxyapatite (HA) is widely used as bone substitute material in repairing bone defects and injuries. However, the high brittleness and low fatigue strength of HA make it difficult to satisfy the long-term service requirements of implants. In general, the mechanical properties of ceramics can be improved by sintering which usually operated above 1000 °C. The high temperature not only brings huge energy consumption and environmental pollution, but also loss the surface-active sites on HA, which greatly limits the biological functions of the material, such as drug loading and degradation. A method that can achieve the densification of HA at a lower temperature is particularly important to improve mechanical and biological properties. In this work, a promising drug-loaded HA-polylactic acid (HA-PLA) composite with high densification and mechanical properties was prepared by cold sintering process at 80 °C, in addition, the relatively low operating temperature enables the material to realize in-situ loading of drugs. The cold-sintered composite with HA/PLA was 7/3 showed the best performance with significant biodegradation (16.8%) and drug release (95%) ability. Therefore, this study provides a simple and universal method for the preparation of biological composite material, which provides a useful platform for efficient in situ drug loading biomaterials.

In situ study on the hierarchical interfacial “global regulation – equilibrium iteration” bearing-toughening mechanisms in Strombus gigas shell by synchrotron radiation computed tomography technique

Strombus gigas shell is a natural biological composite material with excellent strength and toughness, and its excellent performance is closely related to its cross-laminated multistage structure. Studying the damage evolution process to reveal the strengthening mechanism is very important. Through synchrotron radiation computed tomography experiments, the important damage evolution phenomenon from outside to inside of the whole structure, from single interface to multistage interfaces (i.e. first-order, second-order and third-order interfaces), were observed. The first-order interface plays the role of global regulation of stress distribution and failure mode in the whole structure, and the second-order and third-order interfaces play the role of regulating stress distribution during crack propagation. These interfaces cooperate with each other while playing respective roles, showing the optimization and control mechanism of “iterative to balance”. New comprehensions of multistage interfaces are expected to provide new strategies for the optimization design and manufacturing of artificial composite materials.

The information content of tensile tests of human hair (wet) is limited: Variables mainly cluster in just two principal components

Tensile testing of keratin fibres, such as wool and hair, in the wet state is a well established tool in the academic as well as applied portfolio of mechanical analyses. For the tensile curve of a hair fibre, fourteen material-specific stress-, strain, and work-related variables were identified. Analysing twelve samples of cosmetically untreated hair, we show that the variables show good precision and a satisfactory degree of homogeneity across samples. Correlation analysis shows strong relationships between essentially all variables. Some strong correlations were in fact expected. However, their actual extent is very surprising, including even rather dissimilar variables (e.g., elastic-vs break-modulus). This observation led us to investigate whether variables may be clustered into a number of groups in order to define a set of underlying orthogonal factors, using Factor Analysis. The results show that 87% of data variance can be accounted for by just two factors, which we refer to as ‘stress-’ and ‘strain’- factor, respectively. The tensile properties of untreated hair (wet) are thus well described by just any pair of strong variables from each factor, such as elastic modulus and break strain. We hypothesize that similar principles may also be applicable for treated hair. This type of approach and its implications may also have relevance for other epidermal appendages (nails, claws, horn, etc) or even more generally for other biological composite materials, such as skin, tendon and bone.

Effect of Micro- and Macro-Mechanical Characteristics of Jujube Bark on Jujube Girdling Quality

Jujube girdling is an important method to improve the yield and quality of jujube fruit. However, girdling usually causes the tearing of jujube bark, resulting in the slow healing and even death of jujube trees. These problems directly affect the economic benefits of jujube planting. In this paper, in order to reveal the tearing mechanism of jujube bark caused by girdling, and provide the basic mechanical parameters of jujube bark for the research and development of automatic girdling equipment, the structure and mechanical characteristics of jujube bark were studied through theory and experimental methods. The results of microscopic observation showed that jujube bark is a biological composite material mainly composed of the phloem, periderm and epidermis. Furthermore, a mathematical model-based analysis of the growth stress of jujube bark showed that growth stress can be regarded as a biaxial stress state, which can promote the longitudinal tearing of jujube bark during girdling. The tensile test results of jujube bark showed that jujube bark belongs to the category of anisotropic and biologic material. Its longitudinal and transverse mechanical characteristics are significantly different. The maximum longitudinal tensile load was in the range of 105.9~293 N, the longitudinal tensile strength was 16.3 MPa and the elastic modulus was 255.6 MPa. The average transverse tensile load was only 27.9% of the longitudinal tensile load. The transverse tensile strength was 3.5 Mpa and its elastic modulus was 724.14 MPa. The transverse cutting test of jujube bark showed that the average cutting force was 118.2 N and the cutting strength was 9.8 MPa. The results of this study showed that both the growth stress and the anisotropic natural polymer of jujube bark have adverse effects on girdling. Subsequently, issues relating to the selection of appropriate girdling tools and the matters needing attention in the process of the girdling operation were proposed, so as to guide the correct application of girdling technology. In addition, some suggestions on the research and development of girdling tools in the future are put forward.

Improving the geometric accuracy in large-scale additive manufacturing of fungus-like adhesive materials

This paper presents results on measurement, analysis, and improvement of the geometric accuracy, namely the difference between the design and printed shapes, in additive manufacturing (AM) of Fungus-Like Adhesive Materials (FLAM). FLAM is a family of natural biological composite materials comprised of cellulose and chitin. They are 3D printed (3DP) using a large-scale AM system based on material extrusion (ME) principles. FLAM is transported and deposited in a form of a high-viscosity colloid which hardens via evaporative loss of its water content at ambient conditions. Various factors affect the geometric accuracy of 3D printing, namely the pliable nature of the material as well as the characteristics of the AM system. The approach used to reduce geometric errors is based on process parameter control via statistical analysis of objects 3D scanned immediately after being 3D printed. Deviations between the design and printed geometries were used to guide error correction by adjusting the machine path used for 3D printing.

Semi-automated differentiation of insect exo- and endocuticle in X-ray microtomography

One of the most versatile and complex biological composite materials, the insect exoskeleton shows a huge range in biomechanical properties. The cuticle exoskeleton can be differentiated into two main histologically different layers with distinct properties: the outer, more sclerotized exocuticle and inner, softer endocuticle. For most biomechanical research questions, it is of great importance to be able to selectively characterize geometrical features of these layers. However, most conventional preparation methods (cross-sections, histological staining, SEM) require complex and destructive sample preparation, which provides only two-dimensional information. Here, we present a novel, simple staining method using X-ray microtomography to distinguish between exo- and endocuticle in a 3D environment without sample destruction. We illustrate the power of our method using locust (Locusta migratoria) hindleg tibia, a well characterized biomechanical sample. Our method allows an easy and direct measurement of exo- and endocuticle and their respective geometric features. Applying our method will help to understand the biomechanical role of exo- and endocuticle within an insect exoskeleton and will allow us to understand its composition and morphological features in more detail.

Innovative research carried out at the nanocenter of the NRC “Kurchatov Institute” – CRISM “Prometey”

This article presents the results of comprehensive innovative research carried out over the past 15 years at the Nanocenter of the NRC “Kurchatov Institute” – CRISM “Prometey” in the following areas: the creation of coatings based on quasicrystals of the Al-Cu-Fe system, laser synthesis technologies, systems electromagnetic protection of technical equipment and biological objects, structural ceramics and composite materials, technologies for surface modification and magnetron sputtering, obtaining powders by melt spraying, hydrogen and alternative energy.

Tensile and compressive strain evolutions of bovine compact bone under four-point bending fatigue loading

Bones are biological composite materials with multiscale structures. Bone fatigue damage is commonly characterized by an increase in strain that is accompanied by microdamage at different scales. This study investigated the damage evolutions of bone specimens under four-point bending fatigue loading using neutral axis migration. Tensile and compressive strains during the fatigue process were simultaneously measured using a digital image correlation technique. The compressive strain of the bone specimen increased rapidly at first and then proceeded slowly while the tensile strain decreased during fatigue loading. Consequently, the neutral axis shifted downward as the damage accumulated. A positive correlation exists between the downward offset of the neutral axis and the number of cycles. The variation in compressive strain is larger than that in tensile strain in this situation.

Comparison of different protocols for demineralization of cortical bone

Bone is a biological composite material consisting of two main components: collagen and mineral. Collagen is the most abundant protein in vertebrates, which makes it of high clinical and scientific interest. In this paper, we compare the composition and structure of cortical bone demineralized using several protocols: ethylene-diamine-tetraacetic acid (EDTA), formic acid (CH2O2), hydrochloric acid (HCl), and HCl/EDTA mixture. The efficiencies of these four agents were investigated by assessing the remaining mineral quantities and collagen integrity with various experimental techniques. Raman spectroscopy results show that the bone demineralized by the CH2O2 agent has highest collagen quality parameter. The HCl/EDTA mixture removes the most mineral, but it affects the collagen secondary structure as amide II bands are shifted as observed by Fourier transform infrared spectroscopy. Thermogravimetric analysis reveals that HCl and EDTA are most effective in removing the mineral with bulk measurements. In summary, we conclude that HCl best demineralizes bone, leaving the well-preserved collagen structure in the shortest time. These findings guide on the best demineralization protocol to obtain high-quality collagen from bone for clinical and scientific applications.

Study on Mechanical and Thermal Performance of Building Energy-saving Wall with Insulation Materials

Sodium alginate and natural fiber were used as modifiers to prepare sodium alginate natural fiber type biological composites. The mechanical properties were characterized by universal testing machine, and the thermal properties were analyzed by conductivity, The effects of wood fiber and straw fiber content on the comprehensive properties of biological composite materials were evaluated by relevant instruments, and the feasibility of biological composite materials as building energy-saving wall insulation materials was evaluated. The results show that the flexural strength, compressive strength and elastic modulus of the composites increase with the increase of wood fiber content. When the wood fiber content is 100%, the mechanical properties of the sample are the best, the flexural strength is 0.573 MPa, and the compressive strength is 1.410 MPa. The results showed that wood fiber and sodium alginate binder were closely combined and had good wettability. The thermal conductivity of biological composite is 0.078-0.089w / (m · K), which means it has good thermal insula tion performance and can be used as thermal insulation material for building energy-saving wall. The results show that the properties of the composite can be improved by adding a higher proportion of wood fiber and a certain amount of glyoxal crosslinking agent.

Comparative biomechanics of hagfish skins: diversity in material, morphology, and movement

The baggy skins of hagfishes confer whole-body flexibility that enables these animals to tie themselves into knots without injury. The skin’s looseness is produced by a subcutaneous blood sinus that decouples the skin and body core and permits the core to contort dramatically without loading the skin in tension or shear. Hagfish skin represents a biological composite material comparable in strength and stiffness to the conventionally taut skins of other fishes. However, our understanding of hagfish skin is restricted to only one of 78 species: The Pacific hagfish Eptatretus stoutii. To determine if other hagfish share similar characteristics with E. stoutii, we measured material properties and compared histological data sets from the skins of four hagfish species: E. springeri, E. stoutii, Myxine glutinosa, and M. hubbsi. We also compared these material properties data with skins from the American eel, Anguilla rostrata. We subjected skin samples from all species to uniaxial tensile tests in order to measure strength, stiffness, extensibility, and toughness of skins stretched along longitudinal and circumferential axes. We also used a series of equibiaxial tensile tests on skin samples from E. stoutii, M. glutinosa, and A. rostrata to measure stiffness of skins simultaneously strained along both axes. Significant results of uniaxial and biaxial tests show that the skins from Eptatretus are anisotropic, being stiffer in the longitudinal axis, and more extensible than the isotropic skins of Myxine. Skins of A. rostrata were stiffer in the circumferential axis and they were stronger, tougher, and stiffer than all hagfish skins examined. The skins of Eptatretus are histologically distinct from Myxine skins and possess arrays of fibers that stain like muscle. These interspecific differences across hagfish skins show a phylogenetic pattern with knotting kinematics and flexibility; both genera belong to distinct but major subfamilies within the Myxinidae, and Eptatretus is known for creating and manipulating a greater diversity of knotting styles than Myxine.

Second harmonic generation characterization of collagen in whole bone.

Bone is a unique biological composite material made up of a highly structured collagen mesh matrix and mineral deposits. Although mineral provides stiffness, collagen's secondary organization provides a critical role in bone elasticity. Here, we performed polarimetric analysis of bone collagen fibers using second harmonic generation (SHG) imaging to evaluate lamella sheets and collagen fiber integrity in intact cranial bone. Our polarimetric data was fitted to a model accounting for diattenuation, polarization cross-talk, and birefringence. We compared our data to the fitted model and found no significant difference between our polarimetric observation and the representation of these scattering properties up to 70 µm deep. We also observed a loss of resolution as we imaged up to 70 µm deep into bone but a conservation of polarimetric response. Polarimetric SHG allows for the discrimination of collagen lamellar sheet structures in intact bone. Our work could allow for label-free identification of disease states and monitor the efficacy of therapies for bone disorders.

Fiber reorientation in hybrid helicoidal composites

Naturally occurring biological materials with stiff fibers embedded in a ductile matrix are commonly known to achieve excellent balance between stiffness, strength and ductility. In particular, biological composite materials with helicoidal architecture have been shown to exhibit enhanced damage tolerance and increased impact energy absorption. However, the role of fiber reorientation inside the flexible matrix of helicoid composites on their mechanical behaviors have not yet been extensively investigated. In the present work, we introduce a Discontinuous Fiber Helicoid (DFH) composite inspired by both the helicoid microstructure in the cuticle of mantis shrimp and the nacreous architecture of the red abalone shell. We employ 3D printed specimens, analytical models and finite element models to analyze and quantify in-plane fiber reorientation in helicoid architectures with different geometrical features. We also introduce additional architectures, i.e., single unidirectional lamina and mono-balanced architectures, for comparison purposes. Compared with associated mono-balanced architectures, helicoid architectures exhibit less fiber reorientation values and lower values of strain stiffening. The explanation for this difference is addressed in terms of the measured in-plane deformation, due to uniaxial tensile of the laminae, correlated to lamina misorientation with respect to the loading direction and lay-up sequence.

Mechanical properties of Bombyx mori silkworm silk fibre and its corresponding silk fibroin filament: A comparative study

Bombyx mori silkworm silk fibre is recognized as a kind of natural biological composite materials. From the view of composite materials science, we carried out systematic tensile tests to study mechanical properties and their variations along the length of silkworm fibre (namely silk bave, composite material) and its corresponding silk fibroin filament (namely silk brin, reinforcement phase) of three complete cocoon silks. The factors affecting the mechanical properties were further discussed, including the size, sericin weight content and β-sheet content. Our experimental results and statistical analyses with the coefficient of variance and two-parameter Weibull function all revealed the following: First, silk brin displays smaller variations in the diameter and mechanical properties, compared to those of the bave. Second, mechanical properties of sericin (matrix phase in silk fibre) were estimated accordingly with the mixture law of composite materials. Third, strength and toughness of silkworm silks have trends of growth with the decreases in the diameter and sericin weight content, or increase in the β-sheet content. Fourth, the relatively comprehensive database on mechanical properties of silkworm silk (bave and brin) was obtained concurrently. The research might help in better understanding mechanical properties of silk fibres and their future applications.

Anisotropic crack propagation and deformation in dentin observed by four-dimensional X-ray nano-computed tomography

Understanding the cracking behaviour of biological composite materials is of practical importance. This paper presents the first study to track the interplay between crack initiation, microfracture and plastic deformation in three dimensions (3D) as a function of tubule and collagen fibril arrangement in elephant dentin using in situ X-ray nano-computed tomography (nano-CT). A nano-indenter with a conical tip has been used to incrementally indent three test-pieces oriented at 0°, 45° and 70° to the long axis of the tubules (i.e. radial to the tusk). For the 0° sample two significant cracks formed, one of which linked up with microcracks in the axial-radial plane of the tusk originating from the tubules and the other one occurred as a consequence of shear deformation at the tubules. The 70° test-piece was able to bear the greatest loads despite many small cracks forming around the indenter. These were diverted by the microstructure and did not propagate significantly. The 45° test-piece showed intermediate behaviour. In all cases strains obtained by digital volume correlation were well in excess of the yield strain (0.9%), indeed some plastic deformation could even be seen through bending of the tubules. The hoop strains around the conical indenter were anisotropic with the smallest strains correlating with the primary collagen orientation (axial to the tusk) and the largest strains aligned with the hoop direction of the tusk. Statement of SignificanceThis paper presents the first comprehensive study of the anisotropic nature of microfracture, crack propagation and deformation in elephant dentin using time-lapse X-ray nano-computed tomography. To unravel the interplay of collagen fibrils and local deformation, digital volume correlation (DVC) has been applied to map the local strain field while the crack initiation and propagation is tracked in real time. Our results highlight the intrinsic and extrinsic shielding mechanisms and correlate the crack growth behavior in nature to the service requirement of dentin to resist catastrophic fracture. This is of wide interest not just in terms of understanding dentin fracture but also can extend beyond dentin to other anisotropic structural composite biomaterials such as bone, antler and chitin.

Chitosan composites nerve conduits

Traumatic nerve defect affected over 200, 00 people in the US. As it affected so many people, autografts as the golden standard solution have their limitations. Nerve conduits is a new method and the materials for conduits establishing contain biological materials, synthetic materials and composite materials. Among these categories, composite conduits appear better regeneration ability. Also, chitosan demonstrated have the ability to help nerve regeneration. Thus, chitosan composite may be a good choice in the future. This article compares six kinds of chitosan composite: Laminin-chitosan-PLGA, Chitosan-Zinc Oxide Nanocomposite, PCL/Chitosan/Verapamil, Chitosan/ hyaluronic acid conduit, Chitosan/simvastatin/Pluronic F-127 hydrogel and Chitin/ Carboxymethyl (CM)-chitosan by conduit structure, establishment method, sciatic functional index (SFI), electrophysiology and histology data. This article concludes that chitosan-drug-drug delivery system showed the best recovery result.

Deproteinization of Cortical Bone: Effects of Different Treatments.

Bone is a biological composite material having collagen and mineral as its main constituents. In order to better understand the arrangement of the mineral phase in bone, porcine cortical bone was deproteinized using different chemical treatments. This study aims to determine the best method to remove the protein constituent while preserving the mineral component. Chemicals used were H2O2, NaOCl, NaOH, and KOH, and the efficacy of deproteinization treatments was determined by thermogravimetric analysis and Raman spectroscopy. The structure of the residual mineral parts was examined using scanning electron microscopy. X-ray diffraction was used to confirm that the mineral component was not altered by the chemical treatments. NaOCl was found to be the most effective method for deproteinization and the mineral phase was self-standing, supporting the hypothesis that bone is an interpenetrating composite. Thermogravimetric analyses and Raman spectroscopy results showed the preservation of mineral crystallinity and presence of residual organic material after all chemical treatments. A defatting step, which has not previously been used in conjunction with deproteinization to isolate the mineral phase, was also used. Finally, Raman spectroscopy demonstrated that the inclusion of a defatting procedure resulted in the removal of some but not all residual protein in the bone.