Lamellar and fibre bundle mechanics of the annulus fibrosus in bovine intervertebral disc

ArticleThe intervertebral disc is a multicomposite structure, with an outer fibrous ring, the annulus fibrosus, retaining a gel-like core, the nucleus pulposus. The disc presents complex mechanical behaviour, and it is of high importance for spine biomechanics. Advances in multiscale modelling and disc repair raised a need for new quantitative data on the finest details of annulus fibrosus mechanics. In this work we explored inter-lamella and inter-bundle behaviour of the outer annulus using micromechanical testing and second harmonic generation microscopy. Twenty-one intervertebral discs were dissected from cow tails; the nucleus and inner annulus were excised to leave a ring of outer annulus, which was tested in circumferential loading while imaging the tissue’s collagen fibres network with sub-micron resolution. Custom software was developed to determine local tissue strains through image analysis. Inter-bundle linear and shear strains were 5.5 and 2.8 times higher than intra-bundle strains. Bundles tended to remain parallel while rotating under loading, with large slipping between them. Inter-lamella linear strain was almost 3 times the intra-lamella one, but no slipping was observed at the junction between lamellae. This study confirms that outer annulus straining is mainly due to bundles slipping and rotating. Further development of disc multiscale modelling and repair techniques should take into account this modular behaviour of the lamella, rather than considering it as a homogeneous fibre-reinforced matrix.Henry Smith Charit

Inexact Bayesian point pattern matching for linear transformations

PublishedArticleWe introduce a novel Bayesian inexact point pattern matching model that assumes that a linear transformation relates the two sets of points. The matching problem is inexact due to the lack of one-to-one correspondence between the point sets and the presence of noise. The algorithm is itself inexact; we use variational Bayesian approximation to estimate the posterior distributions in the face of a problematic evidence term. The method turns out to be similar in structure to the iterative closest point algorithm.This work was supported by the University of Exeter’s Bridging the Gaps initiative, which was funded by EPSRC award EP/I001433/1 and the collaboration was formed through the Exeter Imaging Network

Investigation of changes in bone density and chemical composition associated with bone marrow oedema-type appearances in magnetic resonance images of the equine forelimb

This is the author accepted manuscript. The final version is available from BMC via the DOI in this record.Background The aetiology of bone marrow oedema-like abnormalities (BMOA) seen on magnetic resonance imaging (MRI) is as yet not fully understood. The current study aimed to investigate the potential of projection radiography and Raman microspectroscopy to provide information regarding the underlying physiological changes associated with BMOA in equine bone samples. Methods MRI was used to assess 65 limbs from 43 horses. A subset of 13 limbs provided 25 samples, 8 with BMOA present and 17 as controls; these were examined with projection radiography to assess bone mineral density and Raman spectroscopy to assess bone composition. Statistical analysis was conducted using SPSS, the relationship between BMOA and age was tested using binary logistic regression, other outcome measures via unpaired t-tests. Results Overall BMOA was found to be associated with locally increased bone density (p = 0.011), suggesting increased bone formation; however, no measurable changes relating to bone remodelling were found, and there were no detectable changes in the chemical composition of bone. Conclusions BMOA is associated with locally increased bone density, without an associated change in the chemical composition of bone, suggesting this is not linked to BMOA. The presence of increased bone density associated with BMOA does appear to suggest that an increased amount of bone formation is occurring in these regions, but as Raman microspectroscopy data do not demonstrate any significant changes in bone chemical composition associated with BMOA, it would appear that the increased bone volume is due to a greater amount of bone being formed rather than an imbalance in relation to bone remodelling. The study provides a proof of principle for the use of Raman microspectroscopy and projection radiography in in vitro studies of BMOA.School of Physics, University of Exete

Three-dimensional arrangement of elastic fibers in the human corneal stroma

This is the final version of the article. Available from the publisher via the DOI in this record.The cornea is the main refracting lens in the eye. As part of the outer tunic it has to be resilient, a property conferred by the organisation of the constituent collagen. It also has to be sufficiently elastic to regain its exact shape when deformed, in order not to distort the retinal image. The basis of this elasticity is not fully understood. The purpose of this study was to characterise in three dimensions the arrangement and distribution of elastic fibers in the human corneal stroma, using serial block face scanning electron microscopy. We have demonstrated that there exists a complex network of elastic fibers that appear to originate in the sclera or limbus. These appear as elastic sheets in the limbus and peripheral cornea immediately above the trabecular meshwork which itself appears to extend above Descemet's membrane in the peripheral stroma. From these sheets, elastic fibers extend into the cornea; moving centrally they bifurcate and trifurcate into narrower fibers and are concentrated in the posterior stroma immediately above Descemet's membrane. We contend that elastic sheets will play an important role in the biomechanical deformation and recovery of the peripheral cornea. The network may also have practical implications for understanding the structural basis behind a number of corneal surgeries.We would like to thank Dr Anthony Hayes and Mr Derek Scarborough for help with the histology presented in this study and Dr Sally Hayes for useful discussions concerning data interpretation. This work was funded by a Programme Grant (503626) from the Medical Research Council (to KMM) and an MRC studentship (to TW). We thank the CTC Eye Bank at Bristol, UK and the HDBR at Newcastle, UK, for supply of human corneas. The authors have no conflicts of interest to declare

Magnetically controlled ferromagnetic swimmers

This is the final version of the article. Available from Springer Nature via the DOI in this record.Microscopic swimming devices hold promise for radically new applications in lab-on-a-chip and microfluidic technology, diagnostics and drug delivery etc. In this paper, we demonstrate the experimental verification of a new class of autonomous ferromagnetic swimming devices, actuated and controlled solely by an oscillating magnetic field. These devices are based on a pair of interacting ferromagnetic particles of different size and different anisotropic properties joined by an elastic link and actuated by an external time-dependent magnetic field. The net motion is generated through a combination of dipolar interparticle gradient forces, time-dependent torque and hydrodynamic coupling. We investigate the dynamic performance of a prototype (3.6 mm) of the ferromagnetic swimmer in fluids of different viscosity as a function of the external field parameters (frequency and amplitude) and demonstrate stable propulsion over a wide range of Reynolds numbers. We show that the direction of swimming has a dependence on both the frequency and amplitude of the applied external magnetic field, resulting in robust control over the speed and direction of propulsion. This paves the way to fabricating microscale devices for a variety of technological applications requiring reliable actuation and high degree of control.This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No. 665440. We also acknowledge support via the EPSRC Centre for Doctoral Training in Metamaterials (Grant No. EP/L015331/1)

Biophotonic tools for probing extracellular matrix mechanics

This is the final version. Available on open access from Elsevier via the DOI in this recordThe complex, hierarchical and heterogeneous biomechanics of the extracellular matrix (ECM) are central to the health of multicellular organisms. Characterising the distribution, dynamics and above all else origins of ECM biomechanics are challenges that have captivated researchers for decades. Recently, a suite of biophotonics techniques have emerged as powerful new tools to investigate ECM biomechanics. In this mini-review, we discuss how the non-destructive, sub-micron resolution imaging capabilities of Raman spectroscopy and nonlinear microscopy are being used to interrogate the biomechanics of thick, living tissues. These high speed, label-free techniques are implemented during mechanical testing, providing unprecedented insight into the compositional and structural response of the ECM to changes in the mechanical environment.Rosetrees Trus

Micro-mechanical damages of needle puncture on bovine annulus fibrosus fibrils studies using polarisation-resolved second harmonic generation (P-SHG) microscopy

This is the author accepted manuscript. The final version is available from Elsevier via the DOI in this recordNeedle injection has been widely used in spinal therapeutic or diagnostic processes, such as discography. The use of needles has been suspected in causing mild disc degeneration which can lead to long-term back pain. However, the localised microscopic damage caused by needles has not been well studied. The local progressive damage on a microscopic level caused by needle punctures on the surface of bovine annulus fibrosus was investigated. Four different sizes of needle were used for the puncture and twenty-nine bovine intervertebral discs were studied. Polarization-resolved second harmonic generation and fluorescent microscopy were used to study the local microscopic structural changes in collagen and cell nuclei due to needle damage. Repeated 70 cyclic loadings at ±5% of axial strain were applied after the needle puncture in order to assess progressive damage caused by the needle. Puncture damage on annulus fibrosus were observed either collagen fibre bundles being pushed aside, being cut through or combination of both with part being lift or pushed in. The progressive damage was found less relevant to the needle size and more progressive damage was only observed using the larger needle. Two distinct populations of collagen, in which one was relatively more organised than the other population, were observed especially after the puncture from skewed distribution of polarisation-SHG analysis. Cell shape was found rounder near the puncture site where collagen fibres were damaged.Henry Smith Foundatio

The hierarchical response of human corneal collagen to load

Fibrillar collagen in the human cornea is integral to its function as a transparent lens of precise curvature, and its arrangement is now well-characterised in the literature. While there has been considerable effort to incorporate fibrillar architecture into mechanical models of the cornea, the mechanical response of corneal collagen to small applied loads is not well understood. In this study the fibrillar and molecular response to tensile load was quantified using small and wide angle X-ray scattering (SAXS/WAXS), and digital image correlation (DIC) photography was used to calculate the local strain field that gave rise to the hierarchical changes. A molecular scattering model was used to calculate the tropocollagen tilt relative to the fibril axis and changes associated with applied strain. Changes were measured in the D-period, molecular tilt and the orientation and spacing of the fibrillar and molecular networks. These measurements were summarised into hierarchical deformation mechanisms, which were found to contribute at varying strains. The change in molecular tilt is indicative of a sub-fibrillar “spring-like” deformation mechanism, which was found to account for most of the applied strain under physiological and near-physiological loads. This deformation mechanism may play an important functional role in tissues rich in fibrils of high helical tilt, such as skin and cartilage

Boron-Doped Diamond Dual-Plate Deep-Microtrench Device for Generator-Collector Sulfide Sensing

This is the author accepted manuscript. The final version is available from Wiley via the DOI in this record.A BDD-BDD dual-plate microtrench electrode with 6μm inter-electrode spacing is investigated using generator-collector electrochemistry and shown to give microtrench depth-dependent sulfide detection down to the μM levels. The effect of the microtrench depth is compared for a "shallow" 44 μm and a "deep" 180μm microtrench and linked to the reduction of oxygen to hydrogen peroxide which interferes with sulfide redox cycling. With a deeper microtrench and a fixed collector potential at -1.4V vs. SCE, two distinct redox cycling potential domains are observed at 0.0V vs. SCE (2-electron) and at 1.1V vs. SCE (6-electron).F. M. and A. J. G. thank EPSRC for financial support (EP/I028706/1)

Microstructural Characterisation of Resistance Artery Remodelling in Diabetes Mellitus

This is the final version. Available on open access from Karger Publishers via the DOI in this recordIntroduction: Microvascular remodelling is a symptom of cardiovascular disease. Despite the mechanical environment being recognised as a major contributor to the remodelling process, it is currently only understood in a rudimentary way. Objective: Amorphological and mechanicalevaluation of the resistance vasculature in health and diabetes mellitus.Methods: The cells and extracellular matrix of human subcutaneous resistance arteriesfrom abdominal fat biopsieswere imagedusing two-photon fluorescence and second harmonic generationat varying transmural pressure.The results informed a two-layer mechanical model.Results: Diabetic resistance arteries reducedin wall area as pressure was increased. This was attributed to the presence of thick, straight collagen fibre bundles that bracedthe outer wall.The abnormal mechanical environment caused theinternal elastic lamina and endothelial and vascular smooth muscle cellarrangementsto twist. Conclusions: Our resultssuggest diabetic microvascular remodelling is likely to be stress-driven, comprisingat least two stages: 1. Laying down of adventitial bracing fibres that limit outward distension, and 2. Deposition of additional collagen in the media, likely due to the significantly altered mechanical environment. This work represents a step towards elucidating the local stress environment of cells, which iscrucial to build accurate models of mechanotransduction in disease.British Heart FoundationMedical Research Council (MRC)National Institute for Health Research (NIHR