Systemically administered zoledronic acid activates locally implanted synthetic hydroxyapatite particles enhancing peri-implant bone formation: A regenerative medicine approach to improve fracture fixation.

Fracture fixation in an ageing population is challenging and fixation failure increases mortality and societal costs. We report a novel fracture fixation treatment by applying a hydroxyapatite (HA) based biomaterial at the bone-implant interface and biologically activating the biomaterial by systemic administration of a bisphosphonate (zoledronic acid, ZA). We first used an animal model of implant integration and applied a calcium sulphate (CaS)/HA biomaterial around a metallic screw in the tibia of osteoporotic rats. Using systemic ZA administration at 2-weeks post-surgery, we demonstrated that the implant surrounded by HA particles showed significantly higher peri­implant bone formation compared to the unaugmented implants at 6-weeks. We then evaluated the optimal timing (day 1, 3, 7 and 14) of ZA administration to achieve a robust effect on peri­implant bone formation. Using fluorescent ZA, we demonstrated that the uptake of ZA in the CaS/HA material was the highest at 3- and 7-days post-implantation and the uptake kinetics had a profound effect on the eventual peri­implant bone formation. We furthered our concept in a feasibility study on trochanteric fracture patients randomized to either CaS/HA augmentation or no augmentation followed by systemic ZA treatment. Radiographically, the CaS/HA group showed signs of increased peri­implant bone formation compared with the controls. Finally, apart from HA, we demonstrated that the concept of biologically activating a ceramic material by ZA could also be applied to ß-tricalcium phosphate. This novel approach for fracture treatment that enhances immediate and long-term fracture fixation in osteoporotic bone could potentially reduce reoperations, morbidity and mortality. STATEMENT OF SIGNIFICANCE: • Fracture fixation in an ageing population is challenging. Biomaterial-based augmentation of fracture fixation devices has been attempted but lack of satisfactory biological response limits their widespread use. • We report the biological activation of locally implanted microparticulate hydroxyapatite (HA) particles placed around an implant by systemic administration of the bisphosphonate zoledronic acid (ZA). The biological activation of HA by ZA enhances peri­implant bone formation. •Timing of ZA administration after HA implantation is critical for optimal ZA uptake and consequently determines the extent of peri­implant bone formation. • We translate the developed concept from small animal models of implant integration to a proof-of-concept clinical study on osteoporotic trochanteric fracture patients. • ZA based biological activation can also be applied to other calcium phosphate biomaterials.

Exploratory neutron tomography of articular cartilage.

OBJECTIVE: To investigate the feasibility of using neutron tomography to gain new knowledge of human articular cartilage degeneration in osteoarthritis (OA). Different sample preparation techniques were evaluated to identify maximum intra-tissue contrast. DESIGN: Human articular cartilage samples from 14 deceased donors (18-75 years, 9 males, 5 females) and 4 patients undergoing total knee replacement due to known OA (all female, 61-75 years) were prepared using different techniques: control in saline, treated with heavy water saline, fixed and treated in heavy water saline, and fixed and dehydrated with ethanol. Neutron tomographic imaging (isotropic voxel sizes from 7.5 to 13.5 µm) was performed at two large scale facilities. The 3D images were evaluated for gradients in hydrogen attenuation as well as compared to images from absorption X-ray tomography, magnetic resonance imaging, and histology. RESULTS: Cartilage was distinguishable from background and other tissues in neutron tomographs. Intra-tissue contrast was highest in heavy water-treated samples, which showed a clear gradient from the cartilage surface to the bone interface. Increased neutron flux or exposure time improved image quality but did not affect the ability to detect gradients. Samples from older donors showed high variation in gradient profile, especially from donors with known OA. CONCLUSIONS: Neutron tomography is a viable technique for specialized studies of cartilage, particularly for quantifying properties relating to the hydrogen density of the tissue matrix or water movement in the tissue.

High-resolution neutron imaging of salt precipitation and water transport in zero-gap CO2 electrolysis.

The electrochemical reduction of CO2 is a pivotal technology for the defossilization of the chemical industry. Although pilot-scale electrolyzers exist, water management and salt precipitation remain a major hurdle to long-term operation. In this work, we present high-resolution neutron imaging (6 µm) of a zero-gap CO2 electrolyzer to uncover water distribution and salt precipitation under application-relevant operating conditions (200 mA cm-2 at a cell voltage of 2.8 V with a Faraday efficiency for CO of 99%). Precipitated salts penetrating the cathode gas diffusion layer can be observed, which are believed to block the CO2 gas transport and are therefore the major cause for the commonly observed decay in Faraday efficiency. Neutron imaging further shows higher salt accumulation under the cathode channel of the flow field compared to the land.

Water management in anion-exchange membrane water electrolyzers under dry cathode operation.

Dry cathode operation is a desired operation mode in anion-exchange membrane water electrolyzers to minimize contamination of the generated hydrogen. However, water management under such operation conditions makes it challenging to maintain reliable performance and durability. Here, we utilize high-resolution in situ neutron imaging (∼6 µm effective resolution) to analyze the water content inside the membrane-electrode-assembly of an anion-exchange membrane water electrolyzer. The ion-exchange capacity (IEC) and thus hydrophilicity of the polymer binder in the cathode catalyst layer is varied to study the influence on water content in the anode (mid IEC, 1.8-2.2 meq. g-1 and high IEC, 2.3-2.6 meq. g-1). The neutron radiographies show that a higher ion-exchange capacity binder allows improved water retention, which reduces the drying-out of the cathode at high current densities. Electrochemical measurements confirm a generally better efficiency for a high IEC cell above 600 mA cm-2. At 1.5 A cm-2 the high IEC has a 100 mV lower overpotential (2.1 V vs. 2.2 V) and a lower high frequency resistance (210 mΩ cm-2 vs. 255 mΩ cm-2), which is believed to be linked to the improved cathode water retention and membrane humidification. As a consequence, the performance stability of the high IEC cell at 1 A cm-2 is also significantly better than that of the mid IEC cell (45 mV h-1 vs. 75 mV h-1).

The Hydration State of Bone Tissue Affects Contrast in Neutron Tomographic Images.

Neutron tomography has emerged as a promising imaging technique for specific applications in bone research. Neutrons have a strong interaction with hydrogen, which is abundant in biological tissues, and they can penetrate through dense materials such as metallic implants. However, in addition to long imaging times, two factors have led to challenges in running in situ mechanical characterization experiments on bone tissue using neutron tomography: 1) the high water content in specimens reduces the visibility of internal trabecular structures; 2) the mechanical properties of bone are dependent on the hydration state of the tissue, with drying being reported to cause increased stiffness and brittleness. This study investigates the possibility of improving image quality in terms of neutron transmission and contrast between material phases by drying and rehydrating in heavy water. Rat tibiae and trabecular bovine bone plugs were imaged with neutron tomography at different hydration states and mechanical testing of the bone plugs was carried out to assess effects of drying and rehydration on the mechanical properties of bone. From analysis of image histograms, it was found that drying reduced the contrast between bone and soft tissue, but the contrast was restored with rehydration. Contrast-to-noise ratios and line profiles revealed that the contrast between bone tissue and background was reduced with increasing rehydration duration but remained sufficient for identifying internal structures as long as no free liquid was present inside the specimen. The mechanical analysis indicated that the proposed fluid exchange protocol had no adverse effects on the mechanical properties.

The effect of high relative humidity on a network of water-sensitive particles (couscous) as revealed by in situ X-ray tomography.

Water significantly influences the mechanical behaviour of all granular materials but none as much as hygroscopic amorphous particles. With sufficiently high water content, particles can swell, agglomerate and their mechanical properties can be reduced, having direct effects on the macroscopic response of the material. In the food and pharmaceutical industry this can cause loss of product functionality. Despite their relevance, very little is known about the microscopic processes that induce these phenomena. Previous studies focused on single particle behaviour, the strength of agglomerated particles and the material flowability, leaving unexplored the link connecting the particle behaviour and the bulk response. This experimental study aims to investigate this aspect with quantitative measurements at both particle and macroscopic scales. A sample of fine couscous is exposed to a high relative humidity (RH) air flow, while being subjected to oedometric conditions, in order to reproduce the storage-silo conditions. In the meantime, X-ray tomographies are acquired continuously and the resulting images are analysed. The designed spatial resolution allows each particle of the sample to be identified and tracked, allowing volumetric evolution to be compared to the properties of the whole sample. The analysis reveals a dilation-compaction macroscopic behaviour, a result of the competition between the particle swelling and the higher deformability as the water content increases. The number, orientations and inter-particle contacts are computed. Their area is related to the applied boundary conditions, and is found to be consistent with the particle swelling and dependent on the applied stress direction.

The scale of a martian hydrothermal system explored using combined neutron and x-ray tomography.

Nakhlite meteorites are igneous rocks from Mars that were aqueously altered ~630 million years ago. Hydrothermal systems on Earth are known to provide microhabitats; knowledge of the extent and duration of these systems is crucial to establish whether they could sustain life elsewhere in the Solar System. Here, we explore the three-dimensional distribution of hydrous phases within the Miller Range 03346 nakhlite meteorite using nondestructive neutron and x-ray tomography to determine whether alteration is interconnected and pervasive. The results reveal discrete clusters of hydrous phases within and surrounding olivine grains, with limited interconnectivity between clusters. This implies that the fluid was localized and originated from the melting of local subsurface ice following an impact event. Consequently, the duration of the hydrous alteration was likely short, meaning that the martian crust sampled by the nakhlites could not have provided habitable environments that could harbor any life on Mars during the Amazonian.

Compact and versatile neutron imaging detector with sub-4µm spatial resolution based on a single-crystal thin-film scintillator.

A large and increasing number of scientific domains pushes for high neutron imaging resolution achieved in reasonable times. Here we present the principle, design and performance of a detector based on infinity corrected optics combined with a crystalline Gd3Ga5O12 : Eu scintillator, which provides an isotropic sub-4 µm true resolution. The exposure times are only of a few minutes per image. This is made possible also by the uniquely intense cold neutron flux available at the imaging beamline NeXT-Grenoble. These comparatively rapid acquisitions are compatible with multiple high quality tomographic acquisitions, opening new venues for in-operando testing, as briefly exemplified here.

Characterisation of Single-Phase Fluid-Flow Heterogeneity Due to Localised Deformation in a Porous Rock Using Rapid Neutron Tomography.

The behaviour of subsurface-reservoir porous rocks is a central topic in the resource engineering industry and has relevant applications in hydrocarbon, water production, and CO2 sequestration. One of the key open issues is the effect of deformation on the hydraulic properties of the host rock and, specifically, in saturated environments. This paper presents a novel full-field data set describing the hydro-mechanical properties of porous geomaterials through in situ neutron and X-ray tomography. The use of high-performance neutron imaging facilities such as CONRAD-2 (Helmholtz-Zentrum Berlin) allows the tracking of the fluid front in saturated samples, making use of the differential neutron contrast between "normal" water and heavy water. To quantify the local hydro-mechanical coupling, we applied a number of existing image analysis algorithms and developed an array of bespoke methods to track the water front and calculate the 3D speed maps. The experimental campaign performed revealed that the pressure-driven flow speed decreases, in saturated samples, in the presence of pre-existing low porosity heterogeneities and compactant shear-bands. Furthermore, the observed complex mechanical behaviour of the samples and the associated fluid flow highlight the necessity for 3D imaging and analysis.

Dual modality neutron and x-ray tomography for enhanced image analysis of the bone-metal interface.

The bone tissue formed at the contact interface with metallic implants, particularly its 3D microstructure, plays a pivotal role for the structural integrity of implant fixation. X-ray tomography is the classical imaging technique used for accessing microstructural information from bone tissue. However, neutron tomography has shown promise for visualising the immediate bone-metal implant interface, something which is highly challenging with x-rays due to large differences in attenuation between metal and biological tissue causing image artefacts. To highlight and explore the complementary nature of neutron and x-ray tomography, proximal rat tibiae with titanium-based implants were imaged with both modalities. The two techniques were compared in terms of visualisation of different material phases and by comparing the properties of the individual images, such as the contrast-to-noise ratio. After superimposing the images using a dedicated image registration algorithm, the complementarity was further investigated via analysis of the dual modality histogram, joining the neutron and x-ray data. From these joint histograms, peaks with well-defined grey value intervals corresponding to the different material phases observed in the specimens were identified and compared. The results highlight differences in how neutrons and x-rays interact with biological tissues and metallic implants, as well as the benefits of combining both modalities. Future refinement of the joint histogram analysis could improve the segmentation of structures and tissues, and yield novel information about specimen-specific properties such as moisture content.

Neutron microtomography to investigate the bone-implant interface-comparison with histological analysis.

Bone properties and especially its microstructure around implants are crucial to evaluate the osseointegration of prostheses in orthopaedic, maxillofacial and dental surgeries. Given the intrinsic heterogeneous nature of the bone microstructure, an ideal probing tool to understand and quantify bone formation must be spatially resolved. X-ray imaging has often been employed, but is limited in the presence of metallic implants, where severe artifacts generally arise from the high attenuation of metals to x-rays. Neutron tomography has recently been proposed as a promising technique to study bone-implant interfaces, thanks to its lower interaction with metals. The aim of this study is to assess the potential of neutron tomography for the characterisation of bone tissue in the vicinity of a metallic implant. A standardised implant with a bone chamber was implanted in rabbit bone. Four specimens were imaged with neutron tomography and subsequently compared to non-decalcified histology to stain soft and mineralised bone tissues, used here as a ground-truth reference. An intensity-based image registration procedure was performed to place the 12 histological slices within the corresponding 3D neutron volume. Significant correlations (p < 0.01) were obtained between the two modalities for the bone-implant contact (BIC) ratio (R = 0.77) and the bone content inside the chamber (R = 0.89). The results indicate that mineralised bone tissue can be reliably detected by neutron tomography. However, theBICratio and bone content were found to be overestimated with neutron imaging, which may be explained by its sensitivity to non-mineralised soft tissues, as revealed by histological staining. This study highlights the suitability of neutron tomography for the analysis of the bone-implant interface. Future work will focus on further distinguishing soft tissues from bone tissue, which could be aided by the adoption of contrast agents.

4D imaging of lithium-batteries using correlative neutron and X-ray tomography with a virtual unrolling technique.

The temporally and spatially resolved tracking of lithium intercalation and electrode degradation processes are crucial for detecting and understanding performance losses during the operation of lithium-batteries. Here, high-throughput X-ray computed tomography has enabled the identification of mechanical degradation processes in a commercial Li/MnO2 primary battery and the indirect tracking of lithium diffusion; furthermore, complementary neutron computed tomography has identified the direct lithium diffusion process and the electrode wetting by the electrolyte. Virtual electrode unrolling techniques provide a deeper view inside the electrode layers and are used to detect minor fluctuations which are difficult to observe using conventional three dimensional rendering tools. Moreover, the 'unrolling' provides a platform for correlating multi-modal image data which is expected to find wider application in battery science and engineering to study diverse effects e.g. electrode degradation or lithium diffusion blocking during battery cycling.

Boron-Based Neutron Scintillator Screens for Neutron Imaging.

In digital neutron imaging, the neutron scintillator screen is a limiting factor of spatial resolution and neutron capture efficiency and must be improved to enhance the capabilities of digital neutron imaging systems. Commonly used neutron scintillators are based on 6LiF and gadolinium oxysulfide neutron converters. This work explores boron-based neutron scintillators because 10B has a neutron absorption cross-section four times greater than 6Li, less energetic daughter products than Gd and 6Li, and lower γ-ray sensitivity than Gd. These factors all suggest that, although borated neutron scintillators may not produce as much light as 6Li-based screens, they may offer improved neutron statistics and spatial resolution. This work conducts a parametric study to determine the effects of various boron neutron converters, scintillator and converter particle sizes, converter-to-scintillator mix ratio, substrate materials, and sensor construction on image quality. The best performing boron-based scintillator screens demonstrated an improvement in neutron detection efficiency when compared with a common 6LiF/ZnS scintillator, with a 125% increase in thermal neutron detection efficiency and 67% increase in epithermal neutron detection efficiency. The spatial resolution of high-resolution borated scintillators was measured, and the neutron tomography of a test object was successfully performed using some of the boron-based screens that exhibited the highest spatial resolution. For some applications, boron-based scintillators can be utilized to increase the performance of a digital neutron imaging system by reducing acquisition times and improving neutron statistics.

What comes NeXT? - High-Speed Neutron Tomography at ILL.

Here, we report on a new record in the acquisition time for fast neutron tomography. With an optimized imaging setup, it was possible to acquire single radiographic projection images with 10 ms and full tomographies with 155 projections images and a physical spatial resolution of 200 µm within 1.5 s. This is about 6.7 times faster than the current record. We used the technique to investigate the water infiltration in the soil with a living lupine root system. The fast imaging setup will be part of the future NeXT instrument at ILL in Grenoble with a great field of possible future applications.