Fine Structural Analysis of Degummed Fibroin Fibers Reveals Its Superior Mechanical Capabilities.

Bombyx mori silk fibroin fibers constitute a class of protein building blocks capable of functionalization and reprocessing into various material formats. The properties of these fibers are typically affected by the intense thermal treatments needed to remove the sericin gum coating layer. Additionally, their mechanical characteristics are often misinterpreted by assuming the asymmetrical cross-sectional area (CSA) as a perfect circle. The thermal treatments impact not only the mechanics of the degummed fibroin fibers, but also the structural configuration of the resolubilized protein, thereby limiting the performance of the resulting silk-based materials. To mitigate these limitations, we explored varying alkali conditions at low temperatures for surface treatment, effectively removing the sericin gum layer while preserving the molecular structure of the fibroin protein, thus, maintaining the hierarchical integrity of the exposed fibroin microfiber core. The precise determination of the initial CSA of the asymmetrical silk fibers led to a comprehensive analysis of their mechanical properties. Our findings indicate that the alkali surface treatment raised the Young's modulus and tensile strength, by increasing the extent of the fibers' crystallinity, by approximately 40 % and 50 %, respectively, without compromising their strain. Furthermore, we have shown that this treatment facilitated further production of high-purity soluble silk protein with rheological and self-assembly characteristics comparable to those of native silk feedstock, initially stored in the animal's silk gland. The developed approaches benefits both the development of silk-based materials with tailored properties and the proper mechanical characterization of asymmetrical fibrous biological materials made of natural building blocks.

Nanocompression of individual multilayered polyhedral nanoparticles.

Inorganic layered materials can form hollow multilayered polyhedral nanoparticles. The size of these multi-wall quasi-spherical structures varies from 4 to 300 nm. These materials exhibit excellent tribological and wear-resisting properties. Measuring and evaluating the stiffness of individual nanoparticle is a non-trivial problem. The current paper presents an in situ technique for stiffness measurements of individual WS(2) nanoparticles which are 80 nm or larger using a high resolution scanning electron microscope (HRSEM). Conducting the experiments in the HRSEM allows elucidation of the compression failure strength and the elastic behavior of such nanoparticles under uniaxial compression.

Improved orthodontic stainless steel wires coated with inorganic fullerene-like nanoparticles of WS(2) impregnated in electroless nickel-phosphorous film.

OBJECTIVE: To reduce friction between orthodontic stainless wires and bracket by coating the wire with nickel-phosphorous electroless film impregnated with inorganic fullerene-like nanoparticles of tungsten disulfide (IF-WS(2)) which are potent dry lubricants. METHODS: Coating was preformed by inserting stainless steel (SS) wires into electroless solutions of nickel-phosphorus (Ni-P) and IF-WS(2). The coated wires were analyzed by SEM (scanning electron microscope) and EDS (energy-dispersive X-ray spectrometer) as well as by tribological tests using a ball-on-flat device. Friction tests simulating archwire functioning of the coated and uncoated wires were carried out by an Instron machine. The adhesion properties of the coated wires after friction were analyzed by a Raman microscope. RESULTS: SEM/EDS analysis of the coated wires showed clear impregnation of the IF-WS(2) nanoparticles in the Ni-P matrix. The friction coefficient measured by the ball-on-flat tribometer was significantly reduced (from 0.25 to 0.08). The friction forces as measured with the Instron on the coated wire were reduced by up to 54% (4.00 N+/-0.19 uncoated vs. 1.85 N+/-0.21 coated). Raman spectra showed that even after extensive friction tests the Ni-P with the IF-WS(2) nanoparticles is attached to the underlying stainless steel wire. CONCLUSIONS: It is proposed that the wires coated with these nanoparticles might offer a novel opportunity to substantially reduce friction during tooth movement. A few tests undertaken to evaluate the toxicity of the fullerene-like nanoparticles have provided indications that they might be biocompatible.

Raman imaging of two orthogonal planes within cortical bone.

The lamellar bone's strength is mainly affected by the organization of its mineralized collagen fibers and material composition. In the present study, Raman microspectroscopic and imaging analyses were employed to study a normal human femoral midshaft bone cube-like specimen with a spatial resolution of approximately 1-2 microm. Identical bone lamellae in both longitudinal and transverse directions were analyzed, which allowed us to separate out orientation and composition dependent Raman lines, depending on the polarization directions. This approach gives information about lamellar bone orientation and variation in bone composition. It is shown that the nu1 PO4 to amide I ratio mainly displays lamellar bone orientation; and nu2 PO4 to amide III and CO3 to nu2 PO4 ratios display variation in bone composition. The nu2 PO4 to amide III ratio is higher in the interstitial bone region, whereas the CO3 to nu2 PO4 ratio has lower values in the same region. The present study provides fresh insights into the organization of a lamellar bone tissue from two orthogonal orientations.

Dentin micro-architecture using harmonic generation microscopy.

OBJECTIVES: We present a novel way to create high-resolution three-dimensional images of tooth dentin by harmonic generation scanning laser microscopy. METHODS: The images were taken using a pulsed infrared laser. Three-dimensional reconstruction enables the visualization of individual tubules and the collagen fibrils mesh around them with an optical resolution of approximately 1 microm. RESULTS: The images show micro-morphological details of the dentinal tubules as well as the collagen fibrils at a depth of up to about 200 microm. The data show that while collagen fibrils are organized in planes perpendicular to the tubules, close to the dentin enamel junction they lie also along the long axis of the tubules. CONCLUSIONS: The unique 3D information opens the opportunity to study the collagen fibril arrangement in relation to the tubule orientation within the dentin matrix, and may be applied to study the micro-morphology of normal versus altered dentin.

Biological armors under impact--effect of keratin coating, and synthetic bio-inspired analogues.

A number of biological armors, such as turtle shells, consist of a strong exoskeleton covered with a thin keratin coating. The mechanical role upon impact of this keratin coating has surprisingly not been investigated thus far. Low-velocity impact tests on the turtle shell reveal a unique toughening phenomenon attributed to the thin covering keratin layer, the presence of which noticeably improves the fracture energy and shell integrity. Synthetic substrate/coating analogues were subsequently prepared and exhibit an impact behavior similar to the biological ones. The results of the present study may improve our understanding, and even future designs, of impact-tolerant structures.

Microstructure-microhardness relations in parallel-fibered and lamellar bone.

Understanding the mechanical function of bone material in relation to its structure is a fascinating but very complicated problem to resolve. Part of the complexity arises from the hierarchical structural organization of bone. Microhardness measurements, initially on relatively simply structured parallel-fibered bone, show a marked anisotropy in three orthogonal directions. This may, in part, be due to the highly anisotropic structure of the basic building block of bone, the mineralized collagen fibril. Microhardness measurements made face-on to the layers of crystals and collagen triple helical molecules, show much lower values than those made edge-on to these layers. Microhardness measurements of the much more complex "rotated-plywood" structure of lamellar bone, reveal the well-known general tendency toward anisotropy in relation to the long axis of the bone. A detailed examination of microhardness-microstructure relations of lamellar bone, however, shows that only in certain orientations can microhardness values be related directly to a specific attribute of the lamellar structure. Clearly, the gradual tilting and rotating of the mineralized collagen fibrils that form this structure produce a material that tends toward having isotropic microhardness properties, even though its basic building block is highly anisotropic. This may be an important structural attribute that allows lamellar bone to withstand a variety of mechanical challenges.

Use of high-performance polyethylene fibres as a reinforcing phase in poly(methylmethacrylate) bone cement.

First results are presented concerning the elastic and ultimate mechanical behaviour of p(MMA) bone cement reinforced with as-received and surface-modified Spectra 900 polyethylene fibres. Even though the surface chemistry and reactivity of the fibres was modified, the surface oxidation and surface grafting treatments of the polyethylene fibres apparently did not significantly affect the mechanical properties of the polyethylene-reinforced p(MMA) bone cement or improve the interfacial bonding. This may be attributed to the rather unfavourable area-to-volume ratio of PE fibres for such treatments, as well as to the necessarily low content of PE fibres in the bone cement which does not allow a clear differentiation between the various samples.

On the relationship between the microstructure of bone and its mechanical stiffness.

A recent study of bone structure shows that the plate-shaped carbonate apatite crystals in individual lamellae are arranged in layers across the lamellae, and that the orientation of these layers are different in alternate lamellae. Based on these findings, a new micromechanical model for the Young's modulus of bone is proposed, which accounts for the anisotropy and geometrical characteristics of the material. The model incorporates the platelet-like geometry of the basic reinforcing unit, the presence of alternating thin and thick lamellae, and the orientations of the crystal platelets in the lamellae. The thin and thick lamellae are modeled as orthotropic composite layers made up of thin rectangular apatite platelets within a collagen matrix, and classical orthotropic elasticity theory is used to calculate the Young's modulus of the lamellae. Bone is viewed as an assembly of such orthotropic lamellae bent into cylindrical structures, and having a constant, alternating angle between successive lamellae. The micromechanical model employs a modified rule-of-mixtures to account for the two types of lamellae. The model provides a curve similar to the published experimental data on the angular dependence of Young's modulus, including a local maximum at an angle between 0 and 90 degrees. A rigorous testing of the model awaits additional experimental data.

Anisotropic mechanical properties of lamellar bone using miniature cantilever bending specimens.

The flexural modulus and work-to-fracture properties of circumferential lamellar bone from baboon tibia are presented, based on experiments with miniature cantilever bending specimens. Data is provided for specimens in three orthogonal directions that show marked anisotropy. The advantages of such miniature specimens (about 150 microm in diameter and 2 mm long) include the possibility of sampling very small volumes within a heterogeneous structure such as osteonal bone, or studying biological materials that are not available in large enough volumes for conventional mechanical analysis.

Mechanical alignment of quasi-one-dimensional nanoparticles.

Randomly oriented rod or rope-like nanoparticles on the surface of an elastomeric substrate are aligned along one direction simply by stretching the substrate. The technique is demonstrated here using single-walled carbon nanotube ropes, and the degree of alignment is assessed by polarized Raman spectroscopy. The alignment is preserved after the particles are removed from the substrate surface, showing that the aligned nanoparticles can be stamped in patterns onto another surface.

The effect of hydration on mechanical anisotropy, topography and fibril organization of the osteonal lamellae.

The effect of hydration on the mechanical properties of osteonal bone, in directions parallel and perpendicular to the bone axis, was studied on three length scales: (i) the mineralized fibril level (~100 nm), (ii) the lamellar level (~6 µm); and (iii) the osteon level (up to ~30 µm).We used a number of techniques, namely atomic force microscopy (AFM), nanoindentation and microindentation. The mechanical properties (stiffness, modulus and/or hardness) have been studied under dry and wet conditions. On all three length scales the mechanical properties under dry conditions were found to be higher by 30-50% compared to wet conditions. Also the mechanical anisotropy, represented by the ratio between the properties in directions parallel and perpendicular to the osteon axis (anisotropy ratio, designated here by AnR), surprisingly decreased somewhat upon hydration. AFM imaging of osteonal lamellae revealed a disappearance of the distinctive lamellar structure under wet conditions. Altogether, these results suggest that a change in mineralized fibril orientation takes place upon hydration.

Elastic and ultimate properties of acrylic bone cement reinforced with ultra-high-molecular-weight polyethylene fibers.

A study of the fracture behavior of poly-(methyl methacrylate) (PMMA) bone cement reinforced with short ultra-high-molecular-weight polyethylene (Spectra 900) fibers is presented. Linear elastic and nonlinear elastic fracture mechanics results indicate that a significant reinforcing effect is obtained at fiber contents as low as 1% by weight, but beyond that concentration a plateau value is reached and the fracture toughness becomes insensitive to fiber content. The flexural strength and modulus are apparently not improved by the incorporation of polyethylene fibers in the acrylic cement, probably because of the presence of voids, the poor mixing practice and the weakness of the fiber/matrix interfacial bond. The present polyethylene/PMMA composite presents several advantages as compared to other composite cements, but overall the mechanical performance of this system resembles that of Kevlar 29/PMMA cement, with a few differences. Scanning electron microscopy reveals characteristic micromechanisms of energy absorption in Spectra 900/PMMA bone cement. A scheme for the strength of random fiber-reinforced composites, which is a simple extension of the Kelly and Tyson model for the strength of unidirectional composites, is presented and discussed. Young's modulus and the fracture toughness results are discussed in the framework of existing theories. More fundamental modeling treatments are needed in terms of fracture micromechanisms to understand and optimize the various mechanical properties with respect to structural parameters and cement preparation technique.

Bending and fracture of compact circumferential and osteonal lamellar bone of the baboon tibia.

Lamellar bone is common among primates, either in the form of extended planar circumferential arrays, or as cylindrically shaped osteons. Osteonal bone generally replaces circumferential lamellar bone with time, and it is therefore of much interest to compare the mechanical properties and fracture behavior of these two forms of lamellar bone. This is, however, difficult as natural specimens of circumferential lamellar bone large enough for standard mechanical tests are not available. We found that as a result of treatment with large doses of alendronate, the lateral sides of the diaphyses of baboon tibia contained fairly extensive regions of circumferential lamellar bone, the structure of which appears to be indistinguishable from untreated lamellar bone. Three-point bending tests were used to determine the elastic and ultimate properties of almost pure circumferential lamellar bone and osteonal bone in four different orientations relative to the tibia long axis. After taking into account the differences in porosity and extent of mineralization of the two bone types, the flexural modulus, bending strength, fracture strain and nominal work-to-fracture properties were similar for the same orientations, with some exceptions. This implies that it is the lamellar structure itself that is mainly responsible for these mechanical properties. The fracture behavior and morphologies of the fracture surfaces varied significantly with orientation in both types of bone. This is related to the microstructure of lamellar bone. Osteonal bone exhibited quite different damage-related behavior during fracture as compared to circumferential lamellar bone. Following fracture the two halves of osteonal bone remained attached whereas in circumferential lamellar bone they separated. These differences could well provide significant adaptive advantages to osteonal bone function.

Lamellar bone: structure-function relations.

The term "bone" refers to a family of materials that have complex hierarchically organized structures. These structures are primarily adapted to the variety of mechanical functions that bone fulfills. Here we review the structure-mechanical relations of one bone structural type, lamellar bone. This is the most abundant type in many mammals, including humans. A lamellar unit is composed of five sublayers. Each sublayer is an array of aligned mineralized collagen fibrils. The orientations of these arrays differ in each sublayer with respect to both collagen fibril axes and crystal layers, such that a complex rotated plywood-like structure is formed. Specific functions for lamellar bone, as opposed to the other bone types, could not be identified. It is therefore proposed that the lamellar structure is multifunctional-the "concrete" of the bone family of materials. Experimentally measured mechanical properties of lamellar bone demonstrate a clear-cut anisotropy with respect to the axis direction of long bones. A comparison of the elastic and ultimate properties of parallel arrays of lamellar units formed in primary bone with cylindrically shaped osteonal structures in secondary formed bone shows that most of the intrinsic mechanical properties are built into the lamellar structure. The major advantages of osteonal bone are its fracture properties. Mathematical modeling of the elastic properties based on the lamellar structure and using a rule-of-mixtures approach can closely simulate the measured mechanical properties, providing greater insight into the structure-mechanical relations of lamellar bone.

Fracture toughness of Kevlar 29/poly(methyl methacrylate) composite materials for surgical implantations.

A study of the fracture behaviour of Kevlar 29 reinforced dental cement is undertaken using both linear elastic and nonlinear elastic fracture mechanics techniques. Results from both approaches--of which the nonlinear elastic is believed to be more appropriate--indicate that a reinforcing effect is obtained for the fracture toughness even at very low fibre content. The flexural strength and modulus are apparently not improved, however, by the incorporation of Kevlar 29 fibres in the PMMA cement, probably because of the presence of voids, the poor fibre/matrix interfacial bonding and unsatisfying cement mixing practice. When compared to other PMMA composite cements, the present system appears to be probably more effective than carbon/PMMA, for example, in terms of fracture toughness. More experimental and analytical work is needed so as to optimize the mechanical properties with respect to structural parameters and cement preparation technique.