Spinal disorders such as back pain incur a substantial societal and economic burden. Unfortunately, there is lack of understanding and treatment of these disorders are further impeded by the inability to assess spinal forces in vivo. The aim of this project is to address this challenge by developing and testing a novel image-driven approach that will assess the forces in an individual's spine in vivo by incorporating information acquired from multimodal imaging (magnetic resonance imaging (MRI) and biplane X-rays) in a subject-specific model. Magnetic resonance and biplane X-ray imaging are used to capture information about the anatomy, tissues, and motion of an individual's spine as they perform a range of everyday activities. This information is then utilised in a subject-specific computational model based on the finite element method to predict the forces in their spine. The project is also utilising novel machine learning algorithms and in vitro, six-axis mechanical testing on human, porcine and bovine samples to develop and test the modelling methods rigorously.Abstract
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Methods
Little information exists when using cell viability assays to evaluate cells within whole tissue, particularly specific types such as the intervertebral disc (IVD). When comparing the reported methodologies and the protocols issued by manufacturers, the processing, working times, and dye concentrations vary significantly, making the assay's reproducibility a costly and time-consuming trial and error process. This study aims to develop a detailed step-by-step cell viability assay protocol for evaluating IVD tissue. IVDs were harvested from bovine tails (n=8) and processed at day 0 and after 7 days of culture. Nucleus pulposus (NP) and the annulus fibrosus (AF) 3 mm cuts were incubated at room temperature (26˚C) with a Viability/Cytotoxicity Kit containing Calcein AM and Ethidium Ethidium homodimer-1 for 2 hr, followed by flash freezing in liquid nitrogen. Thirty µm sections were placed in glass slides and sealed with nail varnish or Antifade Mounting Medium. The IVD tissue was imaged within the next 4h after freezing using an inverted confocal laser-scanning microscope equipped with 488 and 543 nm laser lines. Cell viability at day 0 (NP: 92±9.6 % and AF:80±14.0%) and day 7 (NP: 91±7.9% and AF:76±20%) was successfully maintained and evaluated. The incubation time required is dependent on the working temperatures and tissue thickness. The calcein-AM dye will not be retained in the cells for more than four hours. The specimen preparation and culturing protocol have demonstrated good cell viability at day 0 and after seven days of culture. Processing times and sample preparation play an essential role as the cell viability components in most kits hydrolyse or photobleach quickly. A step-by-step replicable protocol for evaluating the cell viability in IVD will facilitate the evaluation of cell and toxicity-related outcomes of biomechanical testing protocols and IVD regenerative therapies.
250 words max Long polished cemented femoral stems, such as the Exeter Hip Revision stem, are one option available to the revision hip arthroplasty surgeon. When proximal bone stock is compromised, distal fixation is often relied upon for stability of the femoral component. In such circumstances, torsional forces can result in debonding and loosening. This study compared the torsional behaviour of a cemented polished and featureless (plain) stem with cemented, polished stems featuring fins or flutes. Nine torsional tests were carried out on each of these three different stem designs. The finned stem construct was significantly stiffer than the fluted stem (mean 24.5 Nm/deg v 17.5 Nm/deg). The plain stem mean stiffness was less than the featured stems (13 Nm/deg), but wide variability lead to no statistically significant difference. The maximum torque of the finned (30.5 Nm) and fluted stems (29 Nm) was significantly higher than the plain stem (10.5 Nm); with no significance to the difference between the finned and fluted stems. Distal stem features may provide a more reliable and greater resistance to torque in polished, cemented revision hip stems. Finned stem features may also increase the stiffness of the construct. Consideration should thus be given to the incorporation of distal stem features in the design of revision hip stems.
