Simple constitutive model for a cortical bone

This short study presents a simple, one-dimensional constitutive model for the cortical bone with haversian structure. The model is developed within the general framework of the continuum damage theory. The kinetic equation is derived (rather than assumed a priori) through consideration of the irreversible changes of the mesostructure. As a consequence the analytical results closely approximate experimental measurements even though the theory does not introduce a single experimentally unidentifiable material parameter.     

Development of a novel method for the strengthening and toughening of irradiation-sterilized bone allografts

Reconstruction of large skeletal defects is a significant and challenging issue. Bone allografts are often used for such reconstructions. However, sterilizing bone allografts by using γ-irradiation, damages collagen and causes the bone to become weak, brittle and less fatigue resistant. In a previous study, we successfully protected the mechanical properties of human cortical bone by conducting a pre-treatment with ribose, a natural and biocompatible agent. This study focuses on examining possible mechanisms by which ribose might protect the bone. We examined the mechanical properties, crosslinking, connectivity and free radical scavenging potentials of the ribose treatment. Human cortical bone beams were treated with varying concentration of ribose (0.06–1.2 M) and γ-irradiation before testing them in 3-point bending. The connectivity and amounts of crosslinking were determined with Hydrothermal-Isometric-Tension testing and High-Performance-Liquid-Chromatography, respectively. The free radical content was measured using Electron Paramagnetic Resonance. Ribose pre-treatment improved the mechanical properties of irradiation sterilized human bone in a pre-treatment concentration-dependent manner. The 1.2 M pre-treatment provided >100% of ultimate strength of normal controls and protected 76% of the work-to-fracture (toughness) lost in the irradiated controls. Similarly, the ribose pre-treatment improved the thermo-mechanical properties of irradiation-sterilized human bone collagen in a concentration-dependent manner. Greater free radical content and pentosidine content were modified in the ribose treated bone. This study shows that the mechanical properties of irradiation-sterilized cortical bone allografts can be protected by incubating the bone in a ribose solution prior to irradiation.     

Biological behavior of a vascularized cortical bone graft. Experimental study of the rabbit fibula

The study of the biological course of a vascularized osseous graft was performed on 64 fibulae in rabbits. Using radiography, standard histology, tetracyclines markage and study of 47-calcium incorporation as means of control. The time-limits of control are fixed at the 5th, 10th, 15th and 21st day and at the 1st, 2nd, 3rd and 5th month. The results show that if they are vascularized or not, the osseous grafts are not subjected to the rule: everything or nothing: "everything is not living in a vascularized osseous graft and everything is not dying in a conventional osseous graft". As for the good quality of the recipient bed, there is no significant difference between the healing time of those two types of grafts.     

Experimental validation for quantitative protein network models

Cellular responses are the consequence of complex reactions of protein networks. The complexity should ultimately be described by a set of formulas in a quantitative fashion, in which each formula defines the reactions in response to given types of input. However, testing these formulas has not been a simple task because of the lack of appropriate means for experimental validation. 'Reverse-phase' lysate microarrays have been proved to be powerful for such requirements and thus can be a good resource for providing an experimental reference point for the theoretical biology of protein networks.     

Experimental validation of a finite element model of the proximal femur using digital image correlation and a composite bone model

Computational biomechanical models are useful tools for supporting orthopedic implant design and surgical decision making, but because they are a simplification of the clinical scenario they must be carefully validated to ensure that they are still representative. The goal of this study was to assess the validity of the generation process of a structural finite element model of the proximal femur employing the digital image correlation (DIC) strain measurement technique. A finite element analysis model of the proximal femur subjected to gait loading was generated from a CT scan of an analog composite femur, and its predicted mechanical behavior was compared with an experimental model. Whereas previous studies have employed strain gauging to obtain discreet point data for validation, in this study DIC was used for full field quantified comparison of the predicted and experimentally measured strains. The strain predicted by the computational model was in good agreement with experimental measurements, with R(2) correlation values from 0.83 to 0.92 between the simulation and the tests. The sensitivity and repeatability of the strain measurements were comparable to or better than values reported in the literature for other DIC tests on tissue specimens. The experimental-model correlation was in the same range as values obtained from strain gauging, but the DIC technique produced more detailed, full field data and is potentially easier to use. As such, the findings supported the validity of the model generation process, giving greater confidence in the model's predictions, and digital image correlation was demonstrated as a useful tool for the validation of biomechanical models.     

Micromechanical modelling of cortical bone

Cortical bone is a heterogeneous material with a complex hierarchical microstructure. In this work, unit cell finite element models were developed to investigate the effect of microstructural morphology on the macroscopic properties of cortical bone. The effect of lacunar and vascular porosities, percentage of osteonal bone and orientation of the Haversian system on the macroscopic elastic moduli and Poisson's ratios was investigated. The results presented provide relationships for applying more locally accurate material properties to larger scale and whole bone models of varying porosity. Analysis of the effect of the orientation of the Haversian system showed that its effects should not be neglected in larger scale models. This study also provides insight into how microstructural features effect local distributions and cause a strain magnification effect. Limitations in applying the unit cell methodology approach to bone are also discussed.     

Experimental validation of miRNA targets

MicroRNAs are natural, single-stranded, small RNA molecules that regulate gene expression by binding to target mRNAs and suppress its translation or initiate its degradation. In contrast to the identification and validation of many miRNA genes is the lack of experimental evidence identifying their corresponding mRNA targets. The most fundamental challenge in miRNA biology is to define the rules of miRNA target recognition. This is critical since the biological role of individual miRNAs will be dictated by the mRNAs that they regulate. Therefore, only as target mRNAs are validated will it be possible to establish commonalities that will enable more precise predictions of miRNA/mRNA interactions. Currently there is no clear agreement as to what experimental procedures should be followed to demonstrate that a given mRNA is a target of a specific miRNA. Therefore, this review outlines several methods by which to validate miRNA targets. Additionally, we propose that multiple criteria should be met before miRNA target validation should be considered "confirmed."     

Compliance calibration for fracture testing of equine cortical bone

An experimental compliance calibration method for measuring crack length in fracture toughness tests of cortical bone was developed. Calibration tests were conducted on twenty compact type fracture specimens machined from the mid-diaphysis of five pairs of equine third metacarpal bones. Specimens were oriented for crack propagation in a direction transverse to the longitudinal axis of the bone. Specimen compliance was determined from the load vs. crack opening displacement record over a range of crack lengths from 0.48 to 0.75 times the specimen width. The results demonstrate that the compliance calibration method developed for isotropic materials can be used to determine crack length in bone, which is transversely isotropic. However, specimens from lateral and dorsal regions exhibited significantly different compliance calibrations even after differences in elastic modulus were taken into account in the normalized compliance.     

A comparison of methods for measuring cortical bone thickness

Repeated measures of periosteal and endosteal widths of the left second metacarpal from two samples of radiographs were made independently with either a dial caliper or an electronic digitizer connected to a microcomputer. In addition, periosteal and endosteal bone widths of a common sample of radiographs were measured with dial calipers and the electronic digitizer. Dial calipers and an electronic digitizer are both very reliable, but intraobserver errors produced with the use of the digitizer are significantly less than those produced with the use of dial calipers. However, measurements made with the electronic digitizer are systematically larger than corresponding widths measured with dial calipers, but this difference is very small.     

Experimental validation of a fragment library for lead discovery using SPR biosensor technology

A new fragment library for lead discovery has been designed and experimentally validated for use in surface plasmon resonance (SPR) biosensor-based screening. The 930 compounds in the library were selected from 4.6 million commercially available compounds using a series of physicochemical and medicinal chemistry filters. They were screened against 3 prototypical drug targets: HIV-1 protease, thrombin and carbonic anhydrase, and a nontarget: human serum albumin. Compound solubility was not a problem under the conditions used for screening. The high sensitivity of the sensor surfaces allowed the detection of interactions for 35% to 97% of the fragments, depending on the target protein. None of the fragments was promiscuous (i.e., interacted with a stoichiometry ≥5:1 with all 4 proteins), and only 2 compounds dissociated slowly from all 4 proteins. The use of several targets proved valuable since several compounds would have been disqualified from the library on the grounds of promiscuity if fewer target proteins had been used. The experimental procedure allowed an efficient evaluation and exploration of the new fragment library and confirmed that the new library is suitable for SPR biosensor-based screening.     

Visualization of a phantom post-yield deformation process in cortical bone

A prominent opacity is evident in the process zone of notched thin wafers of bone loaded in tension. Being recoverable upon unloading, this opaque zone can be stained only when the sample is under load, unlike the classically reported forms of damage which take up the stain in the unloaded state. Furthermore, despite the stain uptake, microcracks are absent in the stained area examined by high magnification optical microscopy and atomic force microscopy (AFM). Therefore, the size scale and the electric charge of the features involved in the process zone were probed at the submicron level by using a wide range of fluorescent dyes of different molecular weights and charges. It was observed that negatively charged dyes penetrate the process zone and that dyes greater than 10 kDa (about 10–20 nm in size) were unable to label the process zone. Digital image correlation (DIC) measurements indicated that the opacity initiates at about 1% principal strain and the strain accumulates up to 14%. While the opacity was largely recoverable upon unloading, the core regions which experienced large strains had permanent residual strains up to 2%, indicating that the observed deformation phenomenon can be interlocked within bone matrix without the formation of microcracks. Based on the similarity of size and their known affinity for negatively charged species, exposure of mineral nanoplatelets is proposed as prime candidates. Therefore, the deformation process reported here may be associated with debonding of mineral crystals from the neighboring collagen molecules. Overall, post-yield deformation of bone at the micron scale takes place by large strain events which are accommodated in bone matrix by the generation of nanoscale positively charged interfaces.     

Prediction and validation of a mechanism to control the threshold for inhibitory synaptic plasticity

Synaptic plasticity, neuronal activity‐dependent sustained alteration of the efficacy of synaptic transmission, underlies learning and memory. Activation of positive‐feedback signaling pathways by an increase in intracellular Ca²⁺ concentration ([Ca²⁺]_i) has been implicated in synaptic plasticity. However, the mechanism that determines the [Ca²⁺]_i threshold for inducing synaptic plasticity is elusive. Here, we developed a kinetic simulation model of inhibitory synaptic plasticity in the cerebellum, and systematically analyzed the behavior of intricate molecular networks composed of protein kinases, phosphatases, etc. The simulation showed that Ca²⁺/calmodulin‐dependent protein kinase II (CaMKII), which is essential for the induction of synaptic plasticity, was persistently activated or suppressed in response to different combinations of stimuli. The sustained CaMKII activation depended on synergistic actions of two positive‐feedback reactions, CaMKII autophosphorylation and CaMKII‐mediated inhibition of a CaM‐dependent phosphodiesterase, PDE1. The simulation predicted that PDE1‐mediated feedforward inhibition of CaMKII predominantly controls the Ca²⁺ threshold, which was confirmed by electrophysiological experiments in primary cerebellar cultures. Thus, combined application of simulation and experiments revealed that the Ca²⁺ threshold for the cerebellar inhibitory synaptic plasticity is primarily determined by PDE1.     

Fatigue of immature baboon cortical bone

Strain-controlled uniaxial fatigue and monotonic tensile tests were conducted on turned femoral cortical bone specimens obtained from baboons at various ages of maturity. Fatigue loading produced a progressive loss in stiffness and an increase in hysteresis prior to failure, indicating that immature primate cortical bone responds to repeated loading in a fashion similar to that previously observed for adult human cortical bone. Bone fatigue resistance under this strain controlled testing decreased during maturation. Maturation was also associated with an increase in bone dry density, ash fraction and elastic modulus. The higher elastic modulus of more mature bone meant that these specimens were subjected to higher stress levels during testing than more immature bone specimens. Anatomical regions along the femoral shaft exhibited differences in strength and fatigue resistance.     

Anisotropic viscoelastic properties of cortical bone

Relaxation Young's modulus of cortical bone was investigated for two different directions with respect to the longitudinal axis of bone (bone axis, BA): the modulus parallel (P) and normal (N) to the BA. The relaxation modulus was analyzed by fitting to the empirical equation previously proposed for cortical bones, i.e., a linear combination of two Kohlraush-Williams-Watts (KWW) functions (Iyo et al., 2003. Biorheology, submitted): E(t)=E0 (A1 exp[-(t/tau1)beta]+(1-A1) exp[-(t/tau2)gamma]), [0 < A1, beta, gamma < 1], where E0 is the initial modulus value E0. Tau1 and tau2(>tau1) are characteristic times of the relaxation, A1 is the fractional contribution of the fast relaxation (KWW1 process) to the whole relaxation process, and beta and gamma are parameters describing the shape of the relaxation modulus. In both P and N samples, the relaxation modulus was described well by the empirical equation. The KWW1 process of a P sample almost completely coincided with that of an N sample. In the slow process (KWW2 process), there was a difference between the relaxation modulus of a P sample and that of an N sample. The results indicate that the KWW1 process in the empirical equation represents the relaxation in the collagen matrix in bone and that the KWW2 process is related to a higher-order structure of bone that is responsible for the anisotropic mechanical properties of bone.     

Modeling microdamage behavior of cortical bone

Bone is a complex material which exhibits several hierarchical levels of structural organization. At the submicron-scale, the local tissue porosity gives rise to discontinuities in the bone matrix which have been shown to influence damage behavior. Computational tools to model the damage behavior of bone at different length scales are mostly based on finite element (FE) analysis, with a range of algorithms developed for this purpose. Although the local mechanical behavior of bone tissue is influenced by microstructural features such as bone canals and osteocyte lacunae, they are often not considered in FE damage models due to the high computational cost required to simulate across several length scales, i.e., from the loads applied at the organ level down to the stresses and strains around bone canals and osteocyte lacunae. Hence, the aim of the current study was twofold: First, a multilevel FE framework was developed to compute, starting from the loads applied at the whole bone scale, the local mechanical forces acting at the micrometer and submicrometer level. Second, three simple microdamage simulation procedures based on element removal were developed and applied to bone samples at the submicrometer-scale, where cortical microporosity is included. The present microdamage algorithm produced a qualitatively analogous behavior to previous experimental tests based on stepwise mechanical compression combined with in situ synchrotron radiation computed tomography. Our results demonstrate the feasibility of simulating microdamage at a physiologically relevant scale using an image-based meshing technique and multilevel FE analysis; this allows relating microdamage behavior to intracortical bone microstructure.     

Experimental validation of a finite element model of a human cadaveric tibia

Finite element (FE) models of long bones are widely used to analyze implant designs. Experimental validation has been used to examine the accuracy of FE models of cadaveric femurs; however, although convergence tests have been carried out, no FE models of an intact and implanted human cadaveric tibia have been validated using a range of experimental loading conditions. The aim of the current study was to create FE models of a human cadaveric tibia, both intact and implanted with a unicompartmental knee replacement, and to validate the models against results obtained from a comprehensive set of experiments. Seventeen strain rosettes were attached to a human cadaveric tibia. Surface strains and displacements were measured under 17 loading conditions, which consisted of axial, torsional, and bending loads. The tibia was tested both before and after implantation of the knee replacement. FE models were created based on computed tomography (CT) scans of the cadaveric tibia. The models consisted of ten-node tetrahedral elements and used 600 material properties derived from the CT scans. The experiments were simulated on the models and the results compared to experimental results. Experimental strain measurements were highly repeatable and the measured stiffnesses compared well to published results. For the intact tibia under axial loading, the regression line through a plot of strains predicted by the FE model versus experimentally measured strains had a slope of 1.15, an intercept of 5.5 microstrain, and an R(2) value of 0.98. For the implanted tibia, the comparable regression line had a slope of 1.25, an intercept of 12.3 microstrain, and an R(2) value of 0.97. The root mean square errors were 6.0% and 8.8% for the intact and implanted models under axial loads, respectively. The model produced by the current study provides a tool for simulating mechanical test conditions on a human tibia. This has considerable value in reducing the costs of physical testing by pre-selecting the most appropriate test conditions or most favorable prosthetic designs for final mechanical testing. It can also be used to gain insight into the results of physical testing, by allowing the prediction of those variables difficult or impossible to measure directly.     

Two-step wiring plasticity--a mechanism for estrogen-induced rewiring of cortical circuits

Estrogens have been shown to exert powerful effects on cognitive behaviors mediated by several areas of the brain including the cortex. Remodeling of spiny synapses is a key step in the rewiring of neuronal circuitry thought to underlie the processing and storage of information in the forebrain. Whereas estrogen has been shown to regulate synapse structure and function, we are only just starting to understand the molecular and cellular underpinnings of how estrogens can modulate neuronal circuits. Here I will review recent molecular and cellular work that offers a potential mechanism of how estrogen may modulate synapse structure and function of cortical neurons. This mechanism allows cortical neurons to respond to activity-dependent stimuli with greater efficacy in a cellular model termed "Two-Step Wiring Plasticity". This novel form of spine plasticity thus provides insight into how estrogens may modulate the rewiring of neuronal circuits, underlying its ability to influencing cortically based behaviors. This article is part of a Special Issue entitled 'Neurosteroids'.     

Penetration of cutting tool into cortical bone: Experimental and numerical investigation of anisotropic mechanical behaviour

An anisotropic mechanical behaviour of cortical bone and its intrinsic hierarchical microstructure act as protective mechanisms to prevent catastrophic failure due to natural loading conditions; however, they increase the extent of complexity of a penetration process in the case of orthopaedic surgery. Experimental results available in literature provide only limited information about processes in the vicinity of a tool–bone interaction zone. Also, available numerical models the bone-cutting process do not account for material anisotropy or the effect of damage mechanisms. In this study, both experimental and numerical studies were conducted to address these issues and to elucidate the effect of anisotropic mechanical behaviour of cortical bone tissue on penetration of a sharp cutting tool. First, a set of tool-penetration experiments was performed in directions parallel and perpendicular to bone axis. Also, these experiments included bone samples cut from four different cortices to evaluate the effect of spatial variability and material anisotropy on the penetration processes. Distinct deformation and damage mechanisms linked to different microstructure orientations were captured using a micro-lens high-speed camera. Then, a novel hybrid FE model employing a smoothed-particle-hydrodynamic domain embedded into a continuum FE one was developed based on the experimental configuration to characterise the anisotropic deformation and damage behaviour of cortical bone under a penetration process. The results of our study revealed a clear anisotropic material behaviour of the studied cortical bone tissue and the influence of the underlying microstructure. The proposed FE model reflected adequately the experimental results and demonstrated the need for the use of the anisotropic and damage material model to analyse cutting of the cortical-bone tissue.