Development of an image-based technique to examine joint congruency at the elbow

Identifying joint contact in articular joints is important for both the biomechanical investigation of joint mechanics and the study of osteoarthritis. The purpose of this study is to develop a proximity mapping technique to non-invasively determine joint congruency, as a surrogate of joint contact. To illustrate the capabilities of this algorithm, a cadaveric upper extremity was positioned at varying degrees of elbow flexion. This technique was validated using a gold standard experimental casting technique. The pattern of the cast showed an excellent agreement with the generated proximity map using the inter-bone distance algorithm. The results from this study agree with the results of previous studies examining joint contact at the elbow both in the location and in the tracking of the joint contact throughout elbow flexion. Ultimately, this technique will lead to an increased understanding of the effect of malalignment and instability of the joint on contact mechanics.     

Micro-computed tomography image-based evaluation of 3D anisotropy degree of polymer scaffolds

Anisotropy is one of the most meaningful determinants of biomechanical behaviour. This study employs micro-computed tomography (μCT) and image techniques for analysing the anisotropy of regenerative medicine polymer scaffolds. For this purpose, three three-dimensional anisotropy evaluation image methods were used: ellipsoid of inertia (EI), mean intercept length (MIL) and tensor scale (t-scale). These were applied to three patterns (a sphere, a cube and a right prism) and to two polymer scaffold topologies (cylindrical orthogonal pore mesh and spherical pores). For the patterns, the three methods provided good results. Regarding the scaffolds, EI mistook both topologies (0.0158, [ ? 0.5683; 0.6001]; mean difference and 95% confidence interval), and MIL showed no significant differences (0.3509, [0.0656; 0.6362]). T-scale is the preferable method because it gave the best capability (0.3441, [0.1779; 0.5102]) to differentiate both topologies. This methodology results in the development of non-destructive tools to engineer biomimetic scaffolds, incorporating anisotropy as a fundamental property to be mimicked from the original tissue and permitting its assessment by means of μCT image analysis.     

Effect of rapamycin on bone mass and strength in the α2(I)‐G610C mouse model of osteogenesis imperfecta

Osteogenesis imperfecta (OI) is commonly caused by heterozygous type I collagen structural mutations that disturb triple helix folding and integrity. This mutant‐containing misfolded collagen accumulates in the endoplasmic reticulum (ER) and induces a form of ER stress associated with negative effects on osteoblast differentiation and maturation. Therapeutic induction of autophagy to degrade the mutant collagens could therefore be useful in ameliorating the ER stress and deleterious downstream consequences. To test this, we treated a mouse model of mild to moderate OI (α2(I) G610C) with dietary rapamycin from 3 to 8 weeks of age and effects on bone mass and mechanical properties were determined. OI bone mass and mechanics were, as previously reported, compromised compared to WT. While rapamycin treatment improved the trabecular parameters of WT and OI bones, the biomechanical deficits of OI bones were not rescued. Importantly, we show that rapamycin treatment suppressed the longitudinal and transverse growth of OI, but not WT, long bones. Our work demonstrates that dietary rapamycin offers no clinical benefit in this OI model and furthermore, the impact of rapamycin on OI bone growth could exacerbate the clinical consequences during periods of active bone growth in patients with OI caused by collagen misfolding mutations.     

Computed Tomography and Optical Imaging of Osteogenesis-angiogenesis Coupling to Assess Integration of Cranial Bone Autografts and Allografts

A major parameter determining the success of a bone-grafting procedure is vascularization of the area surrounding the graft. We hypothesized that implantation of a bone autograft would induce greater bone regeneration by abundant blood vessel formation. To investigate the effect of the graft on neovascularization at the defect site, we developed a micro–computed tomography (µCT) approach to characterize newly forming blood vessels, which involves systemic perfusion of the animal with a polymerizing contrast agent. This method enables detailed vascular analysis of an organ in its entirety. Additionally, blood perfusion was assessed using fluorescence imaging (FLI) of a blood-borne fluorescent agent. Bone formation was quantified by FLI using a hydroxyapatite-targeted probe and µCT analysis. Stem cell recruitment was monitored by bioluminescence imaging (BLI) of transgenic mice that express luciferase under the control of the osteocalcin promoter. Here we describe and demonstrate preparation of the allograft, calvarial defect surgery, µCT scanning protocols for the neovascularization study and bone formation analysis (including the in vivo perfusion of contrast agent), and the protocol for data analysis.The 3D high-resolution analysis of vasculature demonstrated significantly greater angiogenesis in animals with implanted autografts, especially with respect to arteriole formation. Accordingly, blood perfusion was significantly higher in the autograft group by the 7^th day after surgery. We observed superior bone mineralization and measured greater bone formation in animals that received autografts. Autograft implantation induced resident stem cell recruitment to the graft-host bone suture, where the cells differentiated into bone-forming cells between the 7^th and 10^th postoperative day. This finding means that enhanced bone formation may be attributed to the augmented vascular feeding that characterizes autograft implantation. The methods depicted may serve as an optimal tool to study bone regeneration in terms of tightly bounded bone formation and neovascularization.     

The effect of carbamazepine on bone structure and strength in control and osteogenesis imperfecta (Col1a2 +/p.G610C ) mice

The inherited brittle bone disease osteogenesis imperfecta (OI) is commonly caused by COL1A1 and COL1A2 mutations that disrupt the collagen I triple helix. This causes intracellular endoplasmic reticulum (ER) retention of the misfolded collagen and can result in a pathological ER stress response. A therapeutic approach to reduce this toxic mutant load could be to stimulate mutant collagen degradation by manipulating autophagy and/or ER‐associated degradation. Since carbamazepine (CBZ) both stimulates autophagy of misfolded collagen X and improves skeletal pathology in a metaphyseal chondrodysplasia model, we tested the effect of CBZ on bone structure and strength in 3‐week‐old male OI Col1a2 ^+/p.G610C and control mice. Treatment for 3 or 6 weeks with CBZ, at the dose effective in metaphyseal chondrodysplasia, provided no therapeutic benefit to Col1a2 ^+/p.G610C mouse bone structure, strength or composition, measured by micro‐computed tomography, three point bending tests and Fourier‐transform infrared microspectroscopy. In control mice, however, CBZ treatment for 6 weeks impaired femur growth and led to lower femoral cortical and trabecular bone mass. These data, showing the negative impact of CBZ treatment on the developing mouse bones, raise important issues which must be considered in any human clinical applications of CBZ in growing individuals.     

A Clustering Method for the States of a Markov Chain with an Application to Primate Communication

In some applications clustering problems in stochastic processes arise when the experimenter wants to improve the insight into the structure of the observed system by lumping its states into clusters. As measure of goodness of a cluster solution we choose the rate of predictability of the forthcoming events which can be quantified by means of the entropy of the lumped process. The entropy in turn can be estimated by making use of a Shannon-McMillan-type theorem. We will establish some asymptotic results for the estimate of entropy in the case where the underlying stochastic process is a Markov chain. Our main tool will be a representation of functions of Markov chains which is due to Blackwell (1957). The method proposed here is applied to data on communication processes in monkeys. The object of our analysis will be to find a subgroup structure in the animal group.     

Raman spectroscopy predicts the link between claw keratin and bone collagen structure in a rodent model of oestrogen deficiency

Osteoporosis is a common disease characterised by reduced bone mass and an increased risk of fragility fractures. Low bone mineral density is known to significantly increase the risk of osteoporotic fractures, however, the majority of non-traumatic fractures occur in individuals with a bone mineral density too high to be classified as osteoporotic. Therefore, there is an urgent need to investigate aspects of bone health, other than bone mass, that can predict the risk of fracture. Here, we successfully predicted association between bone collagen and nail keratin in relation to bone loss due to oestrogen deficiency using Raman spectroscopy. Raman signal signature successfully discriminated between ovariectomised rats and their sham controls with a high degree of accuracy for the bone (sensitivity 89%, specificity 91%) and claw tissue (sensitivity 89%, specificity 82%). When tested in an independent set of claw samples the classifier gave 92% sensitivity and 85% specificity. Comparison of the spectral changes occurring in the bone tissue with the changes occurring in the keratin showed a number of common features that could be attributed to common changes in the structure of bone collagen and claw keratin. This study established that systemic oestrogen deficiency mediates parallel structural changes in both the claw (primarily keratin) and bone proteins (primarily collagen). This strengthens the hypothesis that nail keratin can act as a surrogate marker of bone protein status where systemic processes induce changes.