Hip fractures are associated with poor outcomes and high mortality rates of 30%. The current gold standard to measure bone fragility is a Dual-Energy X-ray Absorptiometry (DEXA) scan measuring bone mineral density. Yet DEXA under-diagnoses bone fragility by 50% (1). To combat the burden of bone fragility, this experimental study combined ultrasound (US) with a sophisticated computational algorithm, namely full wave inversion (FWI), to evaluate femoral bone structure.

The aims were to assess the association of bone structure between the proximal femoral diaphysis and femoral neck; secondly to evaluate whether transverse ultrasound could assess bone structure of the proximal femoral diaphysis.

Bone structure of 19 ex vivo human femora was assessed by micro-CT analysis (mean age 88.11 years, male:female 13:6)(Nikon HMXST 225). Using ImageJ/BoneJ, three 10.2mm subsections of proximal diaphysis and femoral neck underwent analysis: the total bone volume fraction, cortical parameters (bone volume fraction, porosity, thickness) and trabecular parameters (porosity, thickness, spacing and connectivity).

A unique US prototype was developed to analyse fifteen femoral diaphyseal subsections using two P4-1 transducers with a novel tomography sequence (Verasonics, Matlab ver R2019a, FWI TRUST protocol). Comparative quantitative analysis of US and Micro-CT measurements was assessed (Graphpad Prism 8.3.1).

Micro-CT analysis of the proximal femoral diaphysis demonstrated low correlation to the femoral neck (Pearson r −0.54 to 0.55). US was able to capture cortical structure, though a wide limit of agreement seen when compared to micro-CT analysis (Bland-Altman range 36–59% difference).

This novel US technique was able to capture cortical bone structure. Improvements in methodology and technology are required to improve the analysis of trabecular bone and overall accuracy. Further evaluation of US and FWI is required to develop the technique and determine its role in clinical practice.

Background

Defining optimal coronal alignment in Total Knee Replacement (TKR) is a controversial and poorly understood subject. Tibial bone density may affect implant stability and functional outcomes following TKR. Our aim was to compare the bone density profile at the implant-tibia interface following TKR in mechanical versus kinematic alignment.

With information about a patient's bone mechanical properties orthopaedic operations could be optimised to reduce intra- and post-operative complications. However, there is currently no reliable method of measuring a patient's bone mechanical properties in vivo. We have previously investigated microindentation, using a 1.5mm diameter spherical indenter tip, and found no correlation between these measurements and compression testing measurements. We hypothesised that by using a larger diameter indenter tip we would closer match bone millimetre-scale mechanical properties.

20 bone samples were taken from 20 patients undergoing hip replacement surgery. The samples were machined from the femoral neck calcar cortical bone into 6×3×3mm parallelepiped specimens, aligned with the osteons along the long axis. The samples were micro-computed tomography (CT) scanned to calculate porosity. Microindentation was performed using a 6mm diameter, sapphire, spherical indenter tip. 12 indentations were performed in a grid and the reduced moduli were calculated using the Oliver-Pharr method. Compression testing was then performed to failure and the apparent elastic modulus was calculated for each sample.

A moderate correlation was found between the indentation reduced moduli and compression testing elastic moduli (r=0.52, r2=0.275, p=0.018). In addition, a moderate correlation was found between the indentation reduced moduli and CT-measured porosity (r=0.5, r2=0.251, p=0.025) and a strong correlation was found between compression testing moduli and porosity (r=0.75, r2=0.568, p<0.001).

Using large-tip spherical microindentation, indentation reduced moduli correlated significantly with compression testing apparent elastic moduli in these 20 cortical bone specimens. Microindentation using a large, spherical indenter tip may predict the mechanical properties of bone at the millimetre length scale and shows promise as a potential future clinical decision aid in surgery.

Tibial bone density may affect implant stability and functional outcomes following total knee replacement (TKR). Our aim was to characterise the bone density profile at the implant-tibia interface following TKR in mechanical versus kinematic alignment. Pre-operative computed tomography scans for 10 patients were obtained. Using surgical planning software, tibial cuts were made for TKR either neutral (mechanical) or 3 degrees varus (kinematic) alignment. Signal intensity, in Hounsfield Units (HU), was measured at 25,600 points throughout an axial slice at the implant-tibia interface and density profiles compared along defined radial axes from the centre of the tibia towards the cortices. From the tibial centre towards the lateral cortex, trabecular bone density for kinematic and mechanical TKR are similar in the inner 50% but differ significantly beyond this (p= 0.012). There were two distinct density peaks, with peak trabecular bone density being higher in kinematic TKR (p<0.001) and peak cortical bone density being higher in mechanical TKR (p<0.01). The difference in peak cortical to peak trabecular signal was 43 HU and 185 HU respectively (p<0.001). On the medial side there was no significant difference in density profile and a linear increase from centre to cortex. In the lateral proximal tibia, peak cortical and peak trabecular bone densities differ between kinematic TKR and mechanical TKR. Laterally, mechanical TKR may be more dependent upon cortical bone for support compared to kinematic TKR, where trabecular bone density is higher. This may have implications for surgical planning and implant design.

Objectives

Bisphosphonates (BP) are the first-line treatment for preventing fragility fractures. However, concern regarding their efficacy is growing because bisphosphonate is associated with over-suppression of remodelling and accumulation of microcracks. While dual-energy X-ray absorptiometry (DXA) scanning may show a gain in bone density, the impact of this class of drug on mechanical properties remains unclear. We therefore sought to quantify the mechanical strength of bone treated with BP (oral alendronate), and correlate data with the microarchitecture and density of microcracks in comparison with untreated controls.

Introduction

The increase in revision joint replacement surgery and fractures of bone around orthopaedic implants may be partly addressed by keeping bone healthy around orthopaedic implants by inserting implants with mechanical properties closer to the patient's bone properties. We do not currently have an accurate way of calculating a patient's bone mechanical properties. We therefore posed a simple question: can data derived from a micro-indenter be used to calculate bone stiffness?

Introduction

Bisphosphonates (BP) are the first-line therapy for preventing osteoporotic fragility fractures. However, concern regarding their efficacy is growing because bisphosphonate use is associated with over-suppression of remodeling. Animal studies have reported that BP therapy is associated with accumulation of micro-cracks (Fig. 1) and a reduction in bone mechanical properties, but the effect on humans has not been investigated. Therefore, our aim was to quantify the mechanical strength of bone treated with BP, and correlate this with the microarchitecture and density of micro-damage in comparison with untreated osteoporotic hip-fractured and non-fractured elderly controls.

Bones are thought to become fragile with advancing age due to a loss of mass and structure. However, there are important aspects of bone fragility and fracture that cannot be explained simply by a loss of bone: 30% of all patients told they have healthy bone based on bone mineral density (BMD) measurements go on to fracture.

It has been suggested that increased fracture risk might also be due to ageing at the nanoscale, which might deteriorate the overall mechanical properties of a bone. However, it is not clear how mechanics at the level of the collagen-mineral matrix relate to mechanical properties of the whole bone, or whether nano-mechanics contribute to fracture risk. In order to answer these questions our group is developing state of the art methods for analysing the structure and function of the collagen mineral matrix under loading.

To image the collagen mineral matrix we obtained beam time on a synchrotron particle accelerator at the Diamond Light Source (Didcot, UK). Electrons are accelerated to near light speed by powerful electromagnets, then slowed to create high energy monochromatic X-Ray beams. Through a combination of X-Ray computed tomography and X-Ray diffraction we have been able to image the collagen/mineral matrix. Furthermore, using in situ loading experiments it has been possible to visualise collagen fibrillar sliding and mineral crystal structure.

Our group is analysing how age related changes in nano-structure affect bone mechanical behaviour. As well as comparing fragility fracture patients with ‘healthy’ age matched controls to investigate whether ageing at the nano-scale could increase fracture risk. We are also assessing the effect of common treatments for bone fragility (e.g. bisphosphonate) on nano-mechanics.

Unfortunately the expense and high radiation dose associated with synchrotron imaging prevents the technology from being adapted for patients. Therefore the next step will be to identify and test tools that can be used to indirectly assess bone chemistry and mechanical properties at point of care (e.g. laser spectroscopy and indentation). The data could be used to improve the diagnosis, monitoring and treatment of bone fragility.

Osteoporosis is a global health issue with 200 million people suffering worldwide and it is a common condition in the elderly. Bisphosphonates including alendronate and risendronate are considered as the first line treatment for osteoporosis. However, there is increasing evidence that bisphosphonate (BP) therapy is associated with atypical fractures. Animal studies have reported a dose-dependent association between the duration of BP therapy and the accumulation of micro-damage. We tested the hypothesis that hip fracture patients treated with BP exhibited greater micro-damage density than untreated fracture and ‘healthy’ aging non-fracture controls.

Trabecular bone cores from patients treated with BP were compared with patients who had not received any treatment for bone metabolic disease (ethics reference: R13004). Non-fractured cadaveric femora from individuals with no history of bone metabolic disease were used as controls. Cores were imaged in high spatial resolution (∼1.3µm) using Synchrotron X-ray tomography (Diamond Light Source Ltd.) A novel classification system was devised to characterise features of micro-damage in the Synchrotron images: micro-cracks, diffuse damage and perforations. Synchrotron micro-CT stacks were visualised and analysed using ImageJ, Avizo and VGStudio MAX.

Our findings show that the BP group had the highest micro-damage density across all groups. The BP group (7.7/mm³) also exhibited greater micro-crack density than the fracture (4.3/mm³) and non-fracture (4.1/mm³) controls. Furthermore, the BP group (1.9/mm³) demonstrated increased diffuse damage when compared to the fracture (0.3/mm³) and non-fracture (0.8/mm³) controls. In contrast, the BP group (1.9mm³) had fewer perforations than fracture (3.0/mm³) and non-fracture controls (3.9/mm³).

BP inhibits bone remodelling, thereby reducing the number of perforated trabeculae, but over-suppression leads to micro-damage accumulation. Accumulated damage could weaken the trabecular bone in the femoral head and neck, increasing the risk of a fracture during a trip or fall.

The increase in revision joint replacement surgery and fractures of bone around orthopaedic implants may be partly addressed by keeping bone healthy around orthopaedic implants by inserting implants with mechanical properties closer to the patient's bone properties. We do not currently have an accurate way of calculating a patient's bone mechanical properties. We are therefore investigating whether microindentation can accurately calculate bone stiffness.

We received ethical approval to retrieve femoral heads and necks from patients undergoing hip replacement surgery for research. Cortical bone from the medial calcar region of the femoral neck was cut into 3×3×6mm cuboid specimens. Micro-indentation testing was performed in the direction of loading of the bone using a MicroMaterials indenter. The samples were kept hydrated and were not fixed or polished. From the unloading curve after indentation, the elastic modulus was calculated, using the Oliver- Pharr method. To assess which microindentation machine settings most precisely calculate the elastic modulus we varied the loading and unloading rates, load and indenter tip shape.

The most precise results were obtained by using a spherical indenter tip (rather than Berkovich tip), high load (10N), a loading rate of 100 mN/s and unloading rate of 300 mN/s with a pause of 60 seconds at maximum load and multiple load cycles with constant loads. Using these settings the mean elastic modulus over 12 cycles of testing was 13.0 GPa (+/- 2.47).

By using a spherical indenter tip and fast unloading it was possible to get precise apparent modulus values. By unloading as fast as possible the effects of bone viscoelastic properties are minimised. By using a spherical indenter tip, plastic deformation at the tip is minimised (compared to the Berkovich tip). We are performing further standard compression tests on the samples to verify the accuracy of the indentation tests.

Ontogeny of long bone cross-sectional geometry has lasting effects on adult bone structure. Growth and development of bone is influenced by biological and mechanical factors but the importance of these factors is poorly understood. A study of prenatal, neonatal and infant development in a bone with simple loading patterns, may improve our understanding. Five vertebral columns aged between 6 months prenatal to 2.5 years postnatal, were analysed to quantify the changes in trabecular architecture before and after birth. Several measures were collected including trabecular: thickness, bone volume fraction, connectivity density, number, structure model index and anisotropy. The findings show that in the first year after birth there is a substantial loss of bone volume via decreasing trabecular thickness and number, which tends to increase after 1.2 years. This sequential pattern of development may be a functional response to the initial requirement for calcium mineral homeostasis before birth, followed by the need for trabecular architecture to adapt to mechanical loading after birth. Calcium is essential for growing neonates and therefore osteoclastic resorbtion is up regulated by increasing parathyroid hormone levels. This may account for the loss of bone between 0–1 year. At one year infants begin to walk bipedally, thus weight bearing and ground reaction forces increase. The stable bone volume and increase in organisation of trabecular architecture after one year may reflect increasing weight bearing and ground reaction forces. These findings suggest that nutritional requirements after birth may have a stronger influence on vertebral trabeculae architecture than learning to walk.

Introduction: In orthopaedic surgery, tendon transfers are used routinely. Examples are the correction of deformities due to spasticity in infantile cerebral palsy or clump foot surgery. Aftertreatment is not evidence based but depends mostly on the surgeons personal preferences. This is especially true for the duration of postoperative immobilization. Adhesions between tendon and surrounding tissue are supposedly a key factor for reduced functionality after immobilisation and it appears to be common sense that the amount of scarring depends on the duration of immobilisation.

The purpose of this study was to determine the optimal (im)mobilisation schema, protecting the suture as well as avoiding impairment of the capability of the transferred tendon to slide properly.

Methods: A tendon transfer of the m. flexor digitorum longus to the dorsal talus was performed in 32 New-Zealand rabbits. The tendon was passed through a drill hole and sutured to itself. Animals were randomised into 6 groups. Groups 1 to 3 experienced mobilisation of varying duration (none, 2 and 4 weeks) after two weeks of immobilisation. Groups 4 to 6 received the same time of mobilisation (4 weeks) but after different periods of immobilisation (4, 6, 8 weeks).

Histomorphological examinations including synovial cell coating, appearance of tendon and tissue interface, inflammation and scarring of the site of surgery were done. The results were analyzed statistically (Kruskal-Wallis-test; Jonckheere Terpstra-test).

Results: Except a difference in development of a synovial cell coating of questionable clinical significance there were no significant findings regarding the histomorphology between the different groups.

Conclusion: In opposition to traditional believes, our results suggest that the influence of postoperative mobilisation or immobilisation towards the formation of scarring and adhesions in tendon transfers may by widely overestimated. This implies that the decision for the duration of postoperative immobilisation should be mainly based on safe ingrowths, without the imminent risk of loss of function.

Instrumented gait analysis has evolved into a widely used tool to define and describe abnormalities of gait. It is used as a tool to enhance the performance in sports as well as to measure the effects of conservative or surgical treatment methods. Patients usually walk very slow during gait training, whereas normal data are obtained at regular walking velocity. This may lead to misinterpretations. The purpose of this study was to determine the effects of walking slow towards gait and to establish normal data for “walking slow” on a treadmill.

10 healthy volunteers with no known gait problem underwent training to accommodate to the conditions of treadmill walking. There were 5 females and 5 males. The mean age was 30 [range 22–56] years. Instrumented gait analysis was performed using a camera system (Motion Analysis Systems). Data obtained were processed by OrthotracTM and the proprietary software of our lab. During data acquisition participants were asked to walk at leisure velocity, then they were asked to slow down as much as possible.

The normal walking velocity of was 0,99 [range 0,78–1,16] m/s. When asked to walk as slow as possible the walking speed decreased to 0,29 [range 0,14–0,50] m/s. We noted a change in the ratio between swing and stance periods with less swing time, as well as a increase of double limb support time. Step length decreased. Changes in the pattern of motion included delayed and increased peak ankle dorsiflexion and decrease of ankle plantar flexion at initial contact. 3-D motion data for hip and knee also demonstrate noteworthy changes, generally resulting in a decrease of joint excursion.

Interpretation of gait data obtained from slow walking patients should consider the effects walking velocity. Locomotion therapy (e.g. for spinal cord injuries) should not force patients into motion patterns that are only found at faster walking velocities.