Freeze casting of hydroxyapatite-titania composites for bone substitutes.

Hydroxyapatite (HA) is commonly used as a bone substitute material, but it lacks mechanical strength when compared to native bone tissues. To improve the efficacy of HA as a bone substitute by improving the mechanical strength and cell growth attributes, porous composite scaffolds of HA and titania (HA-TiO2 ) were fabricated through a freeze-casting process. Three different compositions by weight percent, 25-75 HA-TiO2 , 50-50 HA-TiO2 , and 75-25 HA-TiO2 , were custom-made for testing. After sintering at 1250°C, these composite scaffolds exhibited improved mechanical properties compared to porous HA scaffolds. Substrate mixing was observed, which helped reduce crystal size and introduced new phases such as ß-TCP and CaTiO3 , which also led to improved mechanical properties. The composition of 50-50 HA-TiO2 had the highest ultimate compressive strength of 3.12 ± 0.36 MPa and elastic modulus 63.29 ± 28.75 MPa. Human osteoblast cell proliferation assay also increased on all three different compositions when compared to porous HA at 14 days. These results highlight the potential of freeze casting composites for the fabrication of bone substitutes, which provide enhanced mechanical strength and biocompatibility while maintaining porosity.

Hyphal systems and their effect on the mechanical properties of fungal sporocarps.

Little is known about the mechanical and material properties of hyphae, the single constituent material of Agaricomycetes fungi, despite a growing interest in fungus-based materials. In the Agaricomycetes (the mushrooms and allies), there are three types of hyphae that make up sporocarps: generative, skeletal, and ligative. All filamentous Agaricomycetes can be categorized into one of three categories of hyphal systems that compose them: monomitic, dimitic, and trimitic. Monomitic systems have only generative hyphae. Dimitic systems have generative and either skeletal (most common) or ligative. Trimitic systems are composed of all three kinds of hyphae. SEM imaging, compression testing, and theoretical modeling were used to characterize the material and mechanical properties of representative monomitic, dimitic, and trimitic sporocarps. Compression testing revealed an increase in the compression modulus and compressive strength with the addition of more hyphal types (monomitic to dimitic and dimitic to trimitic). The mesostructure of the trimitic sporocarp was tested and modeled, suggesting that the difference in properties between the solid material and the microtubule mesostructure is a result of differences in structure and not material. Theoretical modeling was completed to estimate the mechanical properties of the individual types of hyphae and showed that skeletal hyphae make the largest contribution to mechanical properties of fungal sporocarps. Understanding the contributions of the different types of hyphae may help in the design and application of fungi-based or bioinspired materials. STATEMENT OF SIGNIFICANCE: This research studies the material and mechanical properties of fungal sporocarps and their hyphae, the single constituent material of Agaricomycetes fungi. Though some work has been done on fungal hyphae, this research studies hyphae in context of the three hyphal systems found in Agaricomycetes fungi and estimates the properties of the hyphal filaments, which has not been done previously. This characterization was performed by analyzing the structures and mechanical properties of fungal sporocarps and calculating the theoretical mechanical properties of their hyphae. This data and the resulting conclusions may lead to a better design and implementation process of fungi-based materials in various applications using the properties now known or calculated.

The melanized layer of Armillaria ostoyae rhizomorphs: Its protective role and functions.

Armillaria ostoyae (Romagn.) Herink is a highly pathogenic fungus that uses exploratory, cordlike structures called rhizomorphs to seek out new sources of nutrition, posing a parasitic threat to natural stands of trees, orchards, and vineyards. Rhizomorphs are notoriously difficult to destroy, and this resilience is due in large part to a melanized layer that protects the rhizomorph. While this structure has been previously observed, its structural and chemical defenses are yet to be discerned. Research was conducted on both lab-cultured and wild-harvested rhizomorph samples. While both environments produce rhizomorphs, only the wild-harvested rhizomorphs produced the melanized layer, allowing for direct investigation of its structure and properties. Imaging, chemical analysis, mechanical testing, and finite element modeling were used to understand the defense mechanisms provided by the melanized layer. Imaging showed a porous outer layer in both types of rhizomorphs, though the pores were smaller in the harvested melanized layer. This melanized layer contained calcium, which provides chemical defense against both human and natural control methods, but was absent from cultured samples. Nanoindentation resulted in a larger variance of hardness values for cultured rhizomorphs than for wild-harvested. Finite element analysis proved that the smaller pore structure of the melanized porous layer had the best balance between maximum deformation and resulting permanent deformation. These results allow for a better understanding of the defenses of this pathogenic fungus, which may lead to better control methods.

Thermo-mechanical characterization of shale using nanoindentation.

Shale can be a potential buffer for high-level radioactive nuclear wastes. To be an effective buffer while subject to waste heat, shale's mechanical response at elevated temperature must be known. Many researchers have experimentally characterized the mechanical behavior of various shales at different length scales in adiabatic conditions. However, its mechanical performance at elevated temperatures at the nano-scale remains unknown. To investigate the temperature dependency of nanomechanical properties of shale, we conducted both experimental and numerical studies. In this study, we measured mechanical and fracture properties of shale, such as hardness, elastic modulus, anisotropy, and fracture toughness from 25 °C up to 300 °C at different bedding planes. Statistical analysis of the results suggests that hardness and fracture toughness significantly increased at temperatures from 100 to 300 °C; while, temperature does not have a significant impact on elastic modulus. Data also shows that the bedding plane orientations have a substantial impact on both mechanical and fracture properties of shale at the nano-scale leading to distinct anisotropic behavior at elevated temperature below 100 °C. Additionally, we numerically investigated the mechanical performance of the shale samples at room temperature to gain an insight into its mechanical response through the thickness. Numerical results were validated against the experimental results, confirming the simulation can be used to predict shale deformation at the nano-scale or potentially be used in multi-scale simulations.

Characterization of porous fluorohydroxyapatite bone-scaffolds fabricated using freeze casting.

With the increasing demand for orthopedic and dental reconstruction surgeries, there comes a shortage of viable bone substitutes. This study was therefore designed to assess the efficacy of porous fluorohydroxyapatite (FHA) as a potential bone substitute. For this, porous FHA scaffolds were fabricated using the freeze casting technique. They were then sintered at 1250, 1350 and, 1450 °C, and microstructural, mechanical, and in vitro properties were analyzed. The microstructure analyses revealed the porosity remained constant within the temperature range. However, the pore size decreased with increasing sintering temperature. The greatest compressive strength and elastic modulus were obtained at 1450 °C, which were 13.5 ± 4.0 MPa and 379 ± 182 MPa, respectively. These are comparable values to human trabecular bone and other porous scaffolds made using hydroxyapatite. This analysis has thus helped to attain an understanding of the mechanical and material properties of freeze-cast FHA scaffolds that have not been presented before. In vitro studies revealed an increasing rate of human osteoblast cell proliferation on freeze-cast FHA scaffolds with increasing sintering temperature, suggesting improved osteogenic properties. Additionally, osteoblasts cells were also shown to proliferate into the interior pores of all freeze-cast FHA scaffolds. These results indicate the potential of porous FHA scaffolds fabricated using the freeze-casting technique to be utilized clinically as bone substitutes.

Biological and bioinspired materials: Structure leading to functional and mechanical performance.

Nature has achieved materials with properties and mechanisms that go far beyond the current know-how of the engineering-materials industry. The remarkable efficiency of biological materials, such as their exceptional properties that rely on weak constituents, high performance per unit mass, and diverse functionalities in addition to mechanical properties, has been mostly attributed to their hierarchical structure. Key strategies for bioinspired materials include formulating the fundamental understanding of biological materials that act as inspiration, correlating this fundamental understanding to engineering needs/problems, and fabricating hierarchically structured materials with enhanced properties accordingly. The vast, existing literature on biological and bioinspired materials can be discussed in terms of functional and mechanical aspects. Through essential representative properties and materials, the development of bioinspired materials utilizes the design strategies from biological systems to innovatively augment material performance for various practical applications, such as marine, aerospace, medical, and civil engineering. Despite the current challenges, bioinspired materials have become an important part in promoting innovations and breakthroughs in the modern materials industry.

Synthesis and characterization of novel calcium phosphate glass-derived cements for vital pulp therapy.

Evaluation of the physicochemical behavior and setting reactions of a novel inorganic pulp capping cement which makes use of the unique corrosion properties of sodium metasilicate (NaSi) glass. NaSi and calcium phosphate (CaP) glass powders were synthesized through a melt-quench method. Cements were created by mixing various amounts of the glasses with deionized water at a powder-to-liquid ratio of 2.5 g mL-1. Working and setting times were measured using the indentation standard ISO 9917-1. Sealing ability was tested by placing set samples of each composition in methylene blue dye solution for 24 h. Set samples were also submerged in phosphate buffered saline and incubated at 37 °C for one week. X-ray diffraction was used to identify mature crystalline phases after incubation. Infrared spectroscopy and scanning electron microscopy were used to characterize cements before and after setting and after incubation. Working and setting times measured in the ranges of 2-5 and 10-25 min, respectively. Working and setting time generally decrease with increased NaSi concentration. Cements with compositions of 25 and 33 wt% NaSi were found to resist the infiltration of dye and maintain their shape. Compositions outside this range absorbed dye and collapsed. Infrared spectroscopy provided insight into the setting mechanism of these cements. After one week in vitro, cements were found to contain crystalline phases matching chemically stable, bioactive phases. The combination of NaSi and CaP glasses has favorable setting behavior, sealing ability, and mature phases for pulp capping while relying on a relatively simple, inorganic composition.

3D Printed Templating of Extrinsic Freeze-Casting for Macro-Microporous Biomaterials.

As with most biological materials, natural bone has hierarchical structure. The microstructural features of compact bone are of various length scales with its porosity consisting of larger osteons (∼100 µm diameter) and vascular channels, as well as smaller lacuna spaces (∼10 µm diameter). In this study, the freeze-casting process, which has been previously used to form biocompatible porous scaffolds (made with hydroxyapatite, HA) has been improved to mimic the intrinsic hierarchical structure of natural bone by implementing an extrinsic 3D printed template. The results of pore characterization showed that this novel combined method of 3D printing and freeze-casting is able to produce porosity at multiple length scales. Nonporous, microporous (created with freeze-casting alone), and macro-micro porous (created with freeze-casting and 3D printed templating) scaffolds were compared as substrates to evaluate cellular activities using osteoblast-like MG63 cell lines. The number of cells oriented parallel to the HA wall structures in the freeze-cast scaffold was found to increase on the microporous and macro-micro porous samples when compare to the nonporous samples, mimicking the natural alignment of the lamella of natural bone. Regarding the cell morphologies, cells on microporous and macro-micro porous samples showed narrowly aligned shapes, whereas those on nonporous samples had polygonal shapes with no discernible orientation. Proliferation and differentiation tests demonstrated that no toxicity or functional abnormalities were found in any of the substrates due to potential chemical and mechanical residues that may have been introduced by the freeze-casting process. Monitoring of the three-dimensional distribution of cells in the scaffolds through microcomputed tomography indicates that the cells were well distributed in the interior pore spaces via the interpenetrating macro-micro pore networks. In summary, we demonstrate this novel approach can create porosity at multiple length scales and is highly favorable in creating a biocompatible, osteoconductive, and structurally hierarchical HA scaffolds for biomedical applications.

A comparative analysis of the avian skull: Woodpeckers and chickens.

Woodpeckers peck at trees without any reported brain injury despite undergoing high impact loads. Amongst the adaptations allowing this is a highly functionalized impact-absorption system consisting of the head, beak, tongue and hyoid bone. This study aims to examine the anatomical structure, composition, and mechanical properties of the skull to determine its potential role in energy absorption and dissipation. An acorn woodpecker and a domestic chicken are compared through micro-computed tomography to analyze and compare two- and three-dimensional bone morphometry. Optical and scanning electron microscopy with energy dispersive X-ray spectroscopy are used to identify the structural and chemical components. Nanoindentation reveals mechanical properties along the transverse cross-section, normal to the direction of impact. Results show two different strategies: the skull bone of the woodpecker shows a relatively small but uniform level of closed porosity, a higher degree of mineralization, and a higher cortical to skull bone ratio. Conversely, the chicken skull bone shows a wide range of both open and closed porosity (volume fraction), a lower degree of mineralization, and a lower cortical to skull bone ratio. This structural difference affects the mechanical properties: the skull bones of woodpeckers are slightly stiffer than those of chickens. Furthermore, the Young's modulus of the woodpecker frontal bone is significantly higher than that of the parietal bone. These new findings may be useful to potential engineered design applications, as well as future work to understand how woodpeckers avoid brain injury.

Synergistic structures from magnetic freeze casting with surface magnetized alumina particles and platelets.

Magnetic freeze casting utilizes the freezing of water, a low magnetic field and surface magnetized materials to make multi-axis strengthened porous scaffolds. A much greater magnetic moment was measured for larger magnetized alumina platelets compared with smaller particles, which indicated that more platelet aggregation occurred within slurries. This led to more lamellar wall alignment along the magnetic field direction during magnetic freeze casting at 75 mT. Slurries with varying ratios of magnetized particles to platelets (0:1, 1:3, 1:1, 3:1, 7:1, 1:0) produced porous scaffolds with different structural features and degrees of lamellar wall alignment. The greatest mechanical enhancement in the magnetic field direction was identified in the synergistic condition with the highest particle to platelet ratio (7:1). Magnetic freeze casting with varying ratios of magnetized anisotropic and isotropic alumina provided insights about how heterogeneous morphologies aggregate within lamellar walls that impact mechanical properties. Fabrication of strengthened scaffolds with multi-axis aligned porosity was achieved without introducing different solid materials, freezing agents or additives. Resemblance of 7:1 particle to platelet scaffold microstructure to wood light-frame house construction is framed in the context of assembly inspiration being derived from both natural and synthetic sources.

Stiff, porous scaffolds from magnetized alumina particles aligned by magnetic freeze casting.

Bone consists of a hard mineral phase and a compliant biopolymer phase resulting in a composite material that is both lightweight and strong. Osteoporosis that degrades spongy bone preferentially over time leads to bone brittleness in the elderly. A porous ceramic material that can mimic spongy bone for a one-time implant provides a potential solution for the future needs of an aging population. Scaffolds made by magnetic freeze casting resemble the aligned porosity of spongy bone. A magnetic field applied throughout freezing induces particle chaining and alignment of lamellae structures between growing ice crystals. After freeze drying to extract the ice and sintering to strengthen the scaffold, cubes from the scaffold center are mechanically compressed along longitudinal (z-axis, ice growth direction) and transverse (y-axis, magnetic field direction) axes. The best alignment of lamellar walls in the scaffold center occurs when applying magnetic freeze casting with the largest particles (350nm) at an intermediate magnetic field strength (75mT), which also agrees with stiffness enhancement results in both z and y-axes. Magnetic moments of different sized magnetized alumina particles help determine the ideal magnetic field strength needed to induce alignment in the scaffold center rather than just at the poles.

A Protocol for Bioinspired Design: A Ground Sampler Based on Sea Urchin Jaws.

Bioinspired design is an emerging field that takes inspiration from nature to develop high-performance materials and devices. The sea urchin mouthpiece, known as the Aristotle's lantern, is a compelling source of bioinspiration with an intricate network of musculature and calcareous teeth that can scrape, cut, chew food and bore holes into rocky substrates. We describe the bioinspiration process as including animal observation, specimen characterization, device fabrication and mechanism bioexploration. The last step of bioexploration allows for a deeper understanding of the initial biology. The design architecture of the Aristotle's lantern is analyzed with micro-computed tomography and individual teeth are examined with scanning electron microscopy to identify the microstructure. Bioinspired designs are fabricated with a 3D printer, assembled and tested to determine the most efficient lantern opening and closing mechanism. Teeth from the bioinspired lantern design are bioexplored via finite element analysis to explain from a mechanical perspective why keeled tooth structures evolved in the modern sea urchins we observed. This circular approach allows for new conclusions to be drawn from biology and nature.

Structural analysis of the tongue and hyoid apparatus in a woodpecker.

UNLABELLED: Woodpeckers avoid brain injury while they peck at trees up to 20Hz with speeds up to 7m/s, undergoing decelerations up to 1200g. Along with the head, beak and neck, the hyoid apparatus (tongue bone and associated soft tissues) is subjected to these high impact forces. The shape of the hyoid apparatus is unusual in woodpeckers and its structure and mechanical properties have not been reported in detail. High-resolution X-ray micro-computed tomography and scanning electron microscopy with energy dispersive X-ray spectroscopy were performed and correlated with nanoindentation mapping. The hyoid apparatus has four distinct bone sections, with three joints between these sections. Nanoindentation results on cross-sectional regions of each bone reveal a previously unreported structure consisting of a stiff core and outer, more compliant shell with moduli of up to 27.4GPa and 8.5GPa, respectively. The bending resistance is low at the posterior section of the hyoid bones, indicating that this region has a high degree of flexibility to absorb impact. These new structural findings can be applied to further studies on the energy dissipation of the woodpecker during its drumming behavior, and may have implications for the design of engineered impact-absorbing structures. STATEMENT OF SIGNIFICANCE: Woodpeckers avoid brain injury while they peck at trees, which results in extreme impact conditions. One common adaptation in woodpeckers is the unusual shape of the elongated tongue, also called the hyoid apparatus. The relationship between the structure and mechanical properties of the bony part of the hyoid apparatus has not been previously reported. A three dimensional model of the bony tongue was developed, and the hardness and stiffness were evaluated. A new type of bone structure, which is opposite of typical skeletal bone structure was found. The combined microstructural and mechanical property analysis indicate possible energy absorption routes for the hyoid apparatus and are applicable to the design of engineered structures.

Reproducibility of ZrO2-based freeze casting for biomaterials.

The processing technique of freeze casting has been intensely researched for its potential to create porous scaffold and infiltrated composite materials for biomedical implants and structural materials. However, in order for this technique to be employed medically or commercially, it must be able to reliably produce materials in great quantities with similar microstructures and properties. Here we investigate the reproducibility of the freeze casting process by independently fabricating three sets of eight ZrO2-epoxy composite scaffolds with the same processing conditions but varying solid loading (10, 15 and 20 vol.%). Statistical analyses (One-way ANOVA and Tukey's HSD tests) run upon measurements of the microstructural dimensions of these composite scaffold sets show that, while the majority of microstructures are similar, in all cases the composite scaffolds display statistically significant variability. In addition, composite scaffolds where mechanically compressed and statistically analyzed. Similar to the microstructures, almost all of their resultant properties displayed significant variability though most composite scaffolds were similar. These results suggest that additional research to improve control of the freeze casting technique is required before scaffolds and composite scaffolds can reliably be reproduced for commercial or medical applications.

Structure and mechanical properties of selected protective systems in marine organisms.

Marine organisms have developed a wide variety of protective strategies to thrive in their native environments. These biological materials, although formed from simple biopolymer and biomineral constituents, take on many intricate and effective designs. The specific environmental conditions that shape all marine organisms have helped modify these materials into their current forms: complete hydration, and variation in hydrostatic pressure, temperature, salinity, as well as motion from currents and swells. These conditions vary throughout the ocean, being more consistent in the pelagic and deep benthic zones while experiencing more variability in the nearshore and shallows (e.g. intertidal zones, shallow bays and lagoons, salt marshes and mangrove forests). Of note, many marine organisms are capable of migrating between these zones. In this review, the basic building blocks of these structural biological materials and a variety of protective strategies in marine organisms are discussed with a focus on their structure and mechanical properties. Finally, the bioinspired potential of these biological materials is discussed.

Structural Design Elements in Biological Materials: Application to Bioinspiration.

Eight structural elements in biological materials are identified as the most common amongst a variety of animal taxa. These are proposed as a new paradigm in the field of biological materials science as they can serve as a toolbox for rationalizing the complex mechanical behavior of structural biological materials and for systematizing the development of bioinspired designs for structural applications. They are employed to improve the mechanical properties, namely strength, wear resistance, stiffness, flexibility, fracture toughness, and energy absorption of different biological materials for a variety of functions (e.g., body support, joint movement, impact protection, weight reduction). The structural elements identified are: fibrous, helical, gradient, layered, tubular, cellular, suture, and overlapping. For each of the structural design elements, critical design parameters are presented along with constitutive equations with a focus on mechanical properties. Additionally, example organisms from varying biological classes are presented for each case to display the wide variety of environments where each of these elements is present. Examples of current bioinspired materials are also introduced for each element.

The armored carapace of the boxfish.

The boxfish (Lactoria cornuta) has a carapace consisting of dermal scutes with a highly mineralized surface plate and a compliant collagen base. This carapace must provide effective protection against predators as it comes at the high cost of reduced mobility and speed. The mineralized hydroxyapatite plates, predominantly hexagonal in shape, are reinforced with raised struts that extend from the center toward the edges of each scute. Below the mineralized plates are non-mineralized collagen fibers arranged in through-the-thickness layers of ladder-like formations. At the interfaces between scutes, the mineralized plates form suture-like teeth structures below which the collagen fibers bridge the gap between neighboring scutes. These sutures are unlike most others as they have no bridging Sharpey's fibers and appear to add little mechanical strength to the overall carapace. It is proposed that the sutured interface either allows for accommodation of the changing pressures of the boxfish's ocean habitat or growth, which occurs without molting or shedding. In both tension and punch testing the mineralized sutures remain relatively intact while most failures occur within the collagen fibers, allowing for the individual scutes to maintain their integrity. This complex structure allows for elevated strength of the carapace through an increase in the stressed area when attacked by predators in both penetrating and crushing modes.

Mechanical properties of suture materials in general and cutaneous surgery.

Comprehensive studies comparing tensile properties of sutures are over 25 years old and do not include recent advances in suture materials. Accordingly, the objective of this article is to investigate the tensile properties of commonly used sutures in cutaneous surgery. Thirteen 3-0 sized modern sutures (four nonabsorbable and nine absorbable) were tensile tested in both straight and knotted configurations according to the procedures outlined by the United States Pharmacopeia. Glycomer 631 was found to have the highest failure load (56.1 N) of unknotted absorbable sutures, while polyglyconate (34.2 N) and glycomer 631 (34.3 N) had the highest failure loads of knotted absorbable sutures. Nylon (30.9 N) and polypropylene (18.9 N) had the greatest failure loads of straight and knotted nonabsorbable sutures, respectively. Polydioxane was found to have the most elongation prior to breakage (144%) of absorbable sutures. Silk (8701 MPa) and rapid polyglactin 910 (9320 MPa) had the highest initial modulus of nonabsorbable and absorbable sutures, respectively. The new data presented in the study provide important information for guiding the selection of suture materials for specific surgeries.