The lateral wall thickness (LWT) in trochanteric femoral fractures is a known predictive factor for postoperative fracture stability. Currently, the AO/OTA classification uses a patient non-specific measure to assess the absolute LWT (aLWT) and distinguish stable A1.3 from unstable A2.1 fractures based on a threshold of 20.5 mm. This approach potentially results in interpatient deviations due to different bone morphologies and consequently variations in fracture stability. Therefore, the aim of this study was to explore whether a patient-specific measure for assessment of the relative LWT (rLWT) results in a more precise threshold for prediction of unstable fractures. Part 1 of the study evaluated 146 pelvic radiographs to assess left-right symmetry with regard to caput-collum-angle (CCD) and total trochanteric thickness (TTT), and used the results to establish the rLWT measurement technique. Part 2 reevaluated 202 patients from a previous study cohort to analyze their rLWT versus aLWT for optimization purposes. Findings in Part 1 demonstrated a bilateral symmetry of the femur regarding both CCD and TTT (p ≥ 0.827) allowing to mirror bone's morphology and geometry from the contralateral intact to the fractured femur. Outcomes in Part 2 resulted in an increased accuracy for the new determined rLWT threshold (50.5%) versus the standard 20.5 mm aLWT threshold, with sensitivity of 83.7% versus 82.7% and specificity 81.3% versus 77.8%, respectively. The novel patient-specific rLWT measure can be based on the contralateral femur anatomy and is a more accurate predictor of a secondary lateral wall fracture in comparison to the conventional aLWT. This study established the threshold of 50.5% rLWT as a reference value for prediction of fracture stability and selection of an appropriate implant for fixation of trochanteric femoral fractures

Introduction. Patient-specific biomechanical modeling using Finite Element Analysis (FEA) is pivotal for understanding the structural health of bones, optimizing surgical procedures, assessing outcomes, and validating medical devices, aligning with guidance issued by standards and regulatory bodies. Accurate mapping of image-to-mesh-material is crucial given bone's heterogeneous composition. This study aims to rigorously assess mesh convergence and evaluate the sensitivity of material grouping strategies in quantifying bone strength. Method. Subject-specific geometry and nonlinear material properties were derived from computed tomography (CT) scan data of one cadaveric human vertebral body. Linear tetrahedral elements with varying edge lengths between 2mm and 0.9mm were then generated to study the mesh convergence. To compare the effectiveness of different grouping strategies, three approaches were used: Modulus Gaping (a user-defined absolute threshold of Young's modulus ranging from 500 MPa to 1 MPa), Percentual Thresholding (relative parameter thresholds ranging from 50% to 1%), and Adaptive clustering (unsupervised k-means-based clustering ranging from 10 to 200 clusters). Adaptive clustering enables a constant number of unique material properties in cross-specimen studies, improving the validity of results. Result. Mesh convergence was evaluated via fracture load and reached at a 1mm mesh size across grouping strategies. All strategies exhibit minimal deviation (within 5%) from individually assigned material parameters, except Modulus Gaping, with a 500 MPa threshold (32% difference). Computational efficiency, measured by runtime, significantly improved with grouping strategies, reducing computational cost by 82 to 94% and unique material count by up to 99%. Conclusion. Different grouping strategies offer comparable mesh convergence, highlighting their potential to reduce computational complexity while maintaining accuracy in the biomechanical modeling of bones and suggesting a more efficient approach than individual element materials. The higher efficiency of FEA may increase its applicability in clinical settings with limited computational resources. Further studies are needed to refine grouping parameters and assess their suitability across different subjects

Wearable inertial sensors can detect abnormal gait associated with knee or hip osteoarthritis (OA). However, few studies have compared sensor-derived gait parameters between patients with hip and knee OA or evaluated the efficacy of sensors suitable for remote monitoring in distinguishing between the two. Hence, our study seeks to examine the differences in accelerations captured by low-frequency wearable sensors in patients with knee and hip OA and classify their gait patterns. We included patients with unilateral hip and knee OA. Gait analysis was conducted using an accelerometer ipsilateral with the affected joint on the lateral distal thighs. Statistical parametric mapping (SPM) was used to compare acceleration signals. The k-Nearest Neighbor (k-NN) algorithm was trained on 80% of the signals' Fourier coefficients and validated on the remaining 20% using 10-fold cross-validation to classify the gait patterns into hip and knee OA. We included 42 hip OA patients (19 females, age 70 [63–78], BMI of 28.3 [24.8–30.9]) and 59 knee OA patients (31 females, age 68 [62–74], BMI of 29.7 [26.3–32.6]). The SPM results indicated that one cluster (12–20%) along the vertical axis had accelerations exceeding the critical threshold of 2.956 (p=0.024). For the anteroposterior axis, three clusters were observed exceeding the threshold of 3.031 at 5–19% (p = 0.0001), 39–54% (p=0.00005), and 88–96% (p = 0.01). Regarding the mediolateral axis, four clusters were identified exceeding the threshold of 2.875 at 0–9% (p = 0.02), 14–20% (p=0.04), 28–68% (p < 0.00001), and 84–100% (p = 0.004). The k-NN model achieved an AUC of 0.79, an accuracy of 80%, and a precision of 85%. In conclusion, the Fourier coefficients of the signals recorded by wearable sensors can effectively discriminate the gait patterns of knee and hip OA. In addition, the most remarkable differences in the time domain were observed along the mediolateral axis

Abstract. 1.0 Objectives. Predictive structural models resulting in a trabecular bone topology closely resembling real bone would be a step toward 3D printing of sympathetic prosthetics. This study modifies an established trabecular bone structural adaptation approach, with the objective of achieving an improved adapted topology, specifically connectivity, compared to CT imaging studies; whilst retaining continuum level mechanical properties consistent with those reported in experimental studies. Strain driven structural adaptation models successfully identify trabecular trajectories, although tend to overpredict connectivity and skew trabecular radii distribution towards the smallest radius included in the adaptation. Radius adaptation of each trabecula is driven by a mechanostat approach with a target strain (1250 µɛ) below which radius is decreased (resorption), and above which radius is increased (apposition). Simulations include a lazy zone, in which neither resorption nor apposition takes place (1000 to 1500 µɛ); and a dead zone (<250 µɛ) in which complete resorption of trabeculae with the smallest included radius takes place. This study assesses the impact of increasing the dead zone threshold from <250 µɛ to <1000 µɛ, the lower limit of the lazy zone. 2.0 Methods. In-silico structural models with an initial connectivity (number of trabeculae connecting at each joint) of 14 were generated using a nearest neighbour approach applied to a random cloud of points. Trabeculae were modelled using circular beams whose radii were adapted in response to normal strains caused by the axial force and bending moments due to a vertical pressure of 1 MPa applied to the top of the lattice, with the bottom of the lattice fixed in the vertical direction. Lattices in which nodes are either able (rigid jointed) or unable (pin jointed) to transmit bending moments were considered. Five virtual samples of each lattice type were used, and each simulation repeated twice: with a dead zone of either <250 µɛ or <1000 µɛ. 3.0 Results. In pin jointed lattices the increase in dead zone threshold resulted in reduction of predicted Young's Modulus from 580 MPa (95% CI [577 MPa, 583 MPa]) to 408 MPa (95% CI [397 MPa, 419 MPa]) whilst in rigid jointed lattices it increased form 839MPa (95% CI [832 MPa, 846 MPa]) to 933 MPa (95% CI [931 MPa, 936 MPa]). Mean connectivity decreased from 10.2 to 5.8 in pin jointed simulations and from 9.6 to 3.8 in fixed joined simulations. Topological studies of trabecular bone CT images report a mean connectivity of around 3.4. Pin jointed lattice mean radius increased from 33mm to 45mm, and rigid jointed lattice mean radius increased from 33mm to 64mm. Prevalence of smallest included radius beams decreased in both. 4.0 Conclusion. Improved in-silico representations of trabecular bone can be achieved in structural adaptions by increasing the dead zone threshold and adopting a bending dominated (rigid jointed) lattice structure. Declaration of Interest. (b) declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported:I declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project

The most important outcome predictor of Legg-Calvé-Perthes disease (LCPD) is the shape of the healed femoral head. However, the deformity of the femoral head is currently evaluated by non-reproducible, categorical, and qualitative classifications. In this regard, recent advances in computer vision might provide the opportunity to automatically detect and delineate the outlines of bone in radiographic images for calculating a continuous measure of femoral head deformity. This study aimed to construct a pipeline for accurately detecting and delineating the proximal femur in radiographs of LCPD patients employing existing algorithms. To detect the proximal femur, the pretrained stateof-the-art object detection model, YOLOv5, was trained on 1580 manually annotated radiographs, validated on 338 radiographs, and tested on 338 radiographs. Additionally, 200 radiographs of shoulders and chests were added to the dataset to make the model more robust to false positives and increase generalizability. The convolutional neural network architecture, U-Net, was then employed to segment the detected proximal femur. The network was trained on 80 manually annotated radiographs using real-time data augmentation to increase the number of training images and enhance the generalizability of the segmentation model. The network was validated on 60 radiographs and tested on 60 radiographs. The object detection model achieved a mean Average Precision (mAP) of 0.998 using an Intersection over Union (IoU) threshold of 0.5, and a mAP of 0.712 over IoU thresholds of 0.5 to 0.95 on the test set. The segmentation model achieved an accuracy score of 0.912, a Dice Coefficient of 0.937, and a binary IoU score of 0.854 on the test set. The proposed fully automatic proximal femur detection and segmentation system provides a promising method for accurately detecting and delineating the proximal femoral bone contour in radiographic images, which is necessary for further image analysis

Abstract. Objectives. Acute compartment syndrome (ACS) is a progressive form of muscle ischaemia that is a surgical emergency and can have detrimental outcomes for patients if not treated optimally. The current problem is that there is no clear diagnostic threshold for ACS or guidance as to when fasciotomies should be performed. A new diagnostic method(s) is necessary to provide real-time information about the extent of muscle ischaemia in ACS. Given that lactic acid is produced by cells through anaerobic respiration, it may be possible to measure H+ ion concentration and to use this as a measure of ischaemia within muscle. Although we are familiar with the key biochemical metabolites involved in ischaemia; and the use of viability dyes in cell culture to distinguish between living or dead cells is well recognised; research has not been undertaken to correlate the biochemical and histological findings of ischaemia in skeletal muscle biopsies. Our primary aim was to investigate the potential for viability dyes to be used on live skeletal muscle biopsies (explants). Our secondary aim was to correlate the intramuscular pH readings with muscle biopsy viability. Methods. Nine euthanised Wistar rats were used. A pH catheter was inserted into one exposed gluteus medius muscles to record real-time pH levels and muscle biopsies were taken from the contralateral gluteus medius at the start of experiment and subsequently at every 0.1 of pH unit drop. Prior to muscle biopsy, the surface of the gluteus medius was painted with a layer of 50µmol/l Brilliant blue FCF solution to facilitate biopsy orientation. A 4mm punch biopsy tool was used to take biopsies. Each muscle biopsy was placed in a base mould filled with 4% ultra-low melting point agarose. The agarose embedded tissue block was sectioned to generate 400 micron thick tissue slices with a vibratome. The tissue slices were then placed in the staining solution with Hoechst 33342, Ethidium homodimer-1 and Calcein am. The tissue slices were imaged with Zeiss LSM880 confocal microscope's Z stack function. A dead muscle control was created by adding TritonX-100 to other tissue slices. For quantitative analyses, the images were analysed in Image J using the selection tool. This permitted individual cells to be identified and the mean grey value of each channel to be defined. Using the dead control, we were able to identify the threshold value for living cells using the Calcein AM channel. Results. Viability dyes, used primarily for cell cultures, can be used with skeletal muscle explants. Our study also showed that despite a significant reduction in tissue pH concentration over time, that almost 100% of muscle cells were still viable at pH 6.0, suggesting that skeletal muscle cells are robust to hypoxic insult in the absence of reperfusion. Conclusions. Viability dyes can be used on skeletal muscle biopsies. Further research investigating the likely associations between direct measured pH using a pH catheter, the concentrations of key cellular metabolic markers, and muscle tissue histology using vitality dyes in response to ischaemia, rather than hypoxia, is warranted. Declaration of Interest. (b) declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported:I declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project

An increase in metal ion levels is seen after implantation of all MoM hip prosthesis due to release from the surface directly, more so during articulation and corrosion of the bearing surfaces. The bearing surfaces in MoM prosthesis consist of cobalt, chromium and molybdenum. Several case-reports of cobalt toxicity due to a MoM prosthesis have been published in the last decade. Cobalt intoxication may lead to a variety of symptoms: neuro-ocular toxicity (tinnitus, vertigo, deafness, blindness, convulsions, headaches and peripheral neuropathy), cardiotoxicity and thyroid toxicity. Nausea, anorexia and unexplained weight loss have been described. Systemic effects from metal ions even with well functioning implants or with ion concentrations lower than those associated with known adverse effects may exist and warrant investigation. The aim of this study is to investigate self-reported systemic complaints in association with cobalt ion concentrations in patients with any type of MoM hip prosthesis. A cohort study was conducted. Patients with both unilateral and bilateral, resurfacing and large head metal on metal total hip arthroplasties were included for the current study. Blood metal ion concentrations (cobalt and chromium) were measured by inductively coupled plasma mass spectrometry (ICP-MS). Based on the known cobalt toxicity symptoms of case-reports and toxicology reports a new non-validated questionnaire was developed. questions were subdivided in general questions/symptoms, vestibular symptoms, neurological symptoms, emotional health and cardio- and thyroid toxicity symptoms. Independent samples T test, Fishers Exact Test and Pearsons (R) correlation were used. Analysis was performed on two groups; a low cobalt ion concentration group and a high cobalt ion concentration group A total of 62 patients, 36 (58%) men and 26 (42%) women, were included with a mean age at surgery of 60.8 ± 9.3 years (41.6 – 78.1) and a mean follow up of 6.3 ± 1.4years (3.7 – 9.6). In these patients a total of 71 prosthesis were implanted: 53 unilateral and 9 bilateral. Of these, 44 were resurfacing and 27 large head metal on metal (LHMoM) total hip arthroplasties. Mean cobalt and chromium ion concentrations were 104 ± 141 nmol/L (9 – 833) and 95 ± 130nmol/L (6 – 592), respectively. Based on the different thresholds (120 – 170 or 220 nmol/L) the low cobalt ion concentration group consisted of 44 (71%), 51 (82%) or 55 (89%) subjects respectively. No differences were found in general characteristics, independently of the threshold. The composite score of vestibular symptoms (vision, hearing, tinnitus, dizziness) was significantly higher (p < .050) in all high cobalt ion concentrations groups, independent of the threshold value This study aimed to detect a trend in self-reported systemic complaints in patients with metal-on-metal hip arthroplasty due to raised cobalt ion concentrations. Vestibular symptoms were more common in high cobalt ion concentration groups independent of the three threshold levels tested. The upper limit of acceptable cobalt ion concentrations remains uncertain. With regards to proactively inquired, self-reported symptoms the threshold where effects may be present could be lower than values currently applied in clinical follow-up. It is unknown what exposure to elevated metal ion concentrations for a longer period of time causes with aging subjects. Further research with a larger cohort and a more standardized questionnaire is necessary to detect previously undiscovered or under-reported effects

The use of mesenchymal stromal cells (MSCs) in regenerative medicine and tissue engineering is well established, given their properties of self-renewal and differentiation. However, several studies have shown that these properties diminish with age, and understanding the pathways involved are important to provide regenerative therapies in an ageing population. In this PRISMA systematic review, we investigated the effects of chronological donor ageing on the senescence of MSCs. We identified 3023 studies after searching four databases including PubMed, Web of Science, Cochrane, and Medline. Nine studies met the inclusion and exclusion criteria and were included in the final analyses. These studies showed an increase in the expression of p21, p53, p16, ROS, and NF- B with chronological age. This implies an activated DNA damage response (DDR), as well as increased levels of stress and inflammation in the MSCs of older donors. Additionally, highlighting the effects of an activated DDR in cells from older donors, a decrease in the expression of proliferative markers including Ki67, MAPK pathway elements, and Wnt/ -catenin pathway elements was observed. Furthermore, we found an increase in the levels of SA- -galactosidase, a specific marker of cellular senescence. Together, these findings support an association between chronological age and MSC senescence. The precise threshold for chronological age where the reported changes become significant is yet to be defined and should form the basis for further scientific investigations. The outcomes of this review should direct further investigations into reversing the biological effects of chronological age on the MSC senescence phenotype

µCT images are commonly analysed to assess changes in bone density and architecture in preclinical murine models. Several platforms provide automated analysis of bone architecture parameters from volumetric regions of interest (ROI). However, segmentation of the regions of subchondral bone to create the volumetric ROIs remains a manual and time-consuming task. This study aimed to develop and evaluate automated pipelines for trabecular bone architecture analysis of mouse proximal tibia subchondral bone. A segmented dataset involving 62 knees (healthy and arthritic) from 10-week male C57BL/6 mice were used to train a U-Net type architecture, with µCT scans (downsampled) input that output segmentation and bone volume density (BV/TV) of the subchondral trabecular bone. Segmentations were upsampled and used in tandem with the original scans (10µ) as input for architecture analysis along with the thresholded trabecular bone. The analysis considered the manually and U-Net segmented ROIs using two available pipelines: the ITKBoneMorphometry library and CTan (SKYSCAN). The analyses included: bone volume (BV), total volume (TV), BV/TV, trabecular number (TbN), trabecular thickness (TbTh), trabecular separation (TbSp), and bone surface density (BSBV). There was good agreement for bone measures between the manual and U-Net pipelines utilizing ITK (R=0.88-0.98) and CTan (R=0.91-0.98). ITK and CTan showed good agreement for BV, TV, BV/TV, TbTh and BSBV (R=0.9-0.98). However, a limited agreement was seen between TbN (R=0.73) and TbSb (R=0.59) due to methodological differences in how spacing is evaluated. This U-Net/ITK pipeline seamlessly automated both segmentation and quantification of the proximal tibia subchondral bone. This automated pipeline allows the analysis of large volumes of data, and its open-source nature may enable the standardization of stereologic analysis of trabecular bone across different research groups

Osteochondral glenoid loss is associated with recurrent shoulder instability. The critical threshold for surgical stabilization is multidimensional and conclusively unknown. The aim of this work was to provide a well- measurable surrogate parameter of an unstable shoulder joint for the frequent anterior-inferior dislocation direction. The shoulder stability ratio (SSR) of 10 paired human cadaveric glenoids was determined in anterior-inferior dislocation direction. Osteochondral defects were simulated by gradually removing osteochondral structures in 5%-stages up to 20% of the intact diameter. The glenoid morphological parameters glenoid depth, concavity gradient, and defect radius were measured at each stage by means of optical motion tracking. Based on these parameters, the osteochondral stability ratio (OSSR) was calculated. Correlation analyses between SSR and all morphological parameters, as well as OSSR were performed. The loss of SSR, concavity gradient, depth and OSSR with increasing defect size was significant (all p<0.001). The loss of SSR strongly correlated with the losses of concavity gradient (PCC = 0.918), of depth (PCC = 0.899), and of OSSR (PCC = 0.949). In contrast, the percentage loss based on intact diameter (defect size) correlated weaker with SSR (PCC=0.687). Small osteochondral defects (≤10%) led to significantly higher SSR decrease in small glenoids (diameter <25mm) compared to large (≥ 25mm) ones (p ≤ 0.009). From a biomechanical perspective, the losses of concavity gradient, glenoid depth and OSSR correlate strong with the loss of SSR. Therefore, especially the loss of glenoidal depth may be considered as a valid and reliable alternative parameter to describe shoulder instability. Furthermore, smaller glenoids are more vulnerable to become unstable in case of small osteochondral loosening. On the other hand, the standardly used percentage defect size based on intact diameter correlates weaker with the magnitude of instability and may therefore not be a valid parameter for judgement of shoulder instability

Tourniquet is a commonly used tool in orthopaedic practice. Incidence of complications is low but if any develops, it is devastating. Transient nerve damage, ischemia or skin burns are the possible tourniquet related complications. There is big variation in practice regarding the limb occlusion pressure. 51 procedures in 50 patients were reviewed retrospectively in our district general hospital. We looked at quality of documentation guided by the BOAST standard (The Safe Use of Intraoperative Tourniquets, published in October 2021). Limb occlusion pressure and ischemic time were analysed. Intra-operative and post-operative notes were reviewed to assess quality of documentation and post-operative complications. Although limb occlusion pressure was above the recommended range in more than 75% of cases, there were no significant complications observed. Two cases only developed transient neuropraxia in common peroneal nerve and median nerve following tibial plateau ORIF and trapeziectomy simultaneously. Tibial ORIF fixation case had prolonged ischemic time (more than 120 minutes) and the limb occlusion pressure for the hand case was above the recommended range. Both have recovered within few days with no long-term consequences. Minimum documentation threshold was not met with regarding tourniquet site condition, method of skin isolation and padding, and exsanguination method. This relatively new standard with no previous similar guidance needs time until it is followed by the health care professionals especially when there is no high incidence of complications related to the use of the tourniquet. However, it is crucial to increase the theatre staff awareness of such standards. This will prevent devastating complications specifically in vulnerable patients. Adjustments to theatre checklist have been suggested to improved documentation. Additionally, local teaching sessions will be delivered to theatre personnel aiming at improving our compliance to this standard

Knee osteoarthritis (KOA) diagnosis is based on symptoms, assessed through questionnaires such as the WOMAC. However, the inconsistency of pain recording and the discrepancy between joint phenotype and symptoms highlight the need for objective biomarkers in KOA diagnosis. To this end, we study relationships among clinical and molecular data in a cohort of women (n=51) with Kellgren-Lawrence grade 2–3 KOA through Support Vector Machine (SVM) and a regulation network model (RNM). Clinical descriptors (i.e., pain catastrophism (CA); depression (DE); functionality (FU); joint pain (JP); rigidity (RI); sensitization (SE); synovitis (SY)) are used to classify patients. A Youden's test is performed for each classifier to determine optimal binarization thresholds for the descriptors. Thresholds are tested against patient stratification according to baseline WOMAC data from the Osteoarthritis Initiative, and the mean accuracy is 0.97. For our cohort, the data used as SVM inputs are KOA descriptors, synovial fluid (SL) proteomic measurements (n=25), and transcription factors (TF) activation obtained from RNM [2] stimulated with the SL measurements. The relative weights after classification reflect input importance. The performance of each classifier is evaluated through AUC-ROC analysis. The best classifier with clinical data is CA (AUC = 0.9), highly influenced by FU and SE, suggesting that kinesophobia is involved in pain perception. With SL input, leptin strongly influences every classifier, suggesting the importance of low-grade inflammation. When TF are used, the mean AUC is limited to 0.608, which can be related to the pleomorphic behaviour of osteoarthritic chondrocytes. Nevertheless, FU has an AUC of 0.7 with strong importance of FOXO downregulation. Though larger and longitudinal cohorts are needed, this unique combination of SVM and RNM shall help to map objectively KOA descriptors. Acknowledgements: Catalan & Spanish governments 2020FI_b00680; STRATO-PID2021126469ob-C21-2, European Commission (MSCA-TN-ETN-2020-Disc4All-955735, ERC-2021-CoG-O-Health-101044828). ICREA Academia

Cartilage diseases have a significant impact on the patient's quality of life and are a heavy burden for the healthcare system. Better understanding, early detection and proper follow-up could improve quality of life and reduce healthcare related costs. Therefore, the aim of this study was to evaluate if difference between osteoarthritic (OA) and non-osteoarthritic (non-OA) knees can be detected quantitatively on cartilage and subchondral bone levels with advanced but clinical available imaging techniques. Two OA (mean age = 88.3 years) and three non-OA (mean age = 51.0 years) human cadaveric knees were scanned two times. A high-resolution peripheral quantitative computed tomography (HR-pQCT) scan (XtremeCT, Scanco Medical AG, Switzerland) was performed to quantify the bone microstructure. A contrast-enhanced clinical CT scan (GE Revolution Evo, GE Medical Systems AG, Switzerland) was acquired with the contrast agent Visipaque 320 (60 ml) to measure cartilage. Subregions dividing the condyle in four parts were identified semi-automatically and the images were segmented using adaptive thresholding. Microstructural parameters of subchondral bone and cartilage thickness were quantified. The overall cartilage thickness was reduced by 0.27 mm between the OA and non-OA knees and the subchondral bone quality decreased accordingly (reduction of 33.52 % in BV/TV in the layer from 3 to 8 mm below the cartilage) for the femoral medial condyle. The largest differences were observed at the medial part of the femoral medial condyle both for cartilage and for bone parameters, corresponding to clinical observations. Subchondral bone microstructural parameters and cartilage thickness were quantified using in vivo available imaging and apparent differences between the OA and non-OA knees were detected. Those results may improve OA follow-up and diagnosis and could lead to a better understanding of OA. However, further in vivo studies are needed to validate these methods in clinical practice

We have developed a novel technique to analyse bone, using imaging mass cytometry (IMC) without the constraints of using immunofluorescent histochemistry. IMC can measure the expression of over 40 proteins simultaneously, without autofluorescence. We analysed mitochondrial respiratory chain (RC) protein deficiencies in human bone which are thought to contribute to osteoporosis with increasing age. Osteoporosis is characterised by reduced bone mineral density (BMD) and fragility fractures. Humans accumulate mitochondrial mutations and RC deficiency with age and this has been linked to the changing phenotype in advancing age and age-related disease. Mitochondrial mutations are detectable from the age of 30 onwards, coincidently the age BMD begins to decline. Mitochondria contain their own genome which accumulates somatic variants at around 10 times the rate of nuclear DNA. Once these mutations exceed a threshold, RC deficiency and cellular dysfunction occur. The PolgD257A/D257A mouse model expresses a proof-reading deficient version of PolgA, a mtDNA polymerase. These mice accumulate mutations 3-5 times higher than wild-type mice showing enhanced levels of age-related osteoporosis and RC deficiency in osteoblasts. Bone samples were analysed from young and old patients, developing a protocol and analysis framework for IMC in bone tissue sections to analyse osteoblasts in-situ for RC deficiency. Samples from the femoral neck of 10 older healthy volunteers aged 40 – 85 were compared with samples from young patients aged 1-19. We have identified RC complex I defect in osteoblasts from 6 of the older volunteers, complex II defects in 2 of the older volunteers, complex IV defect in just 1 older volunteer, and complex V defect in 4 of the older volunteers. These observations are consistent with the PolgD257A/D257A mouse-model and suggest that RC deficiency, due to age-related pathogenic mitochondrial DNA mutations, may play a significant role in the pathogenesis of human age-related osteoporosis

Chondral defects in the knee have cartilage biomechanical differences due to defect size and orientation. This study examines how the tibiofemoral contact pressure is affected by increasing full-thickness chondral defect size on the medial and lateral condyle at full extension. Isolated full-thickness, square chondral defects increasing from 0.09cm. 2. to 1.0cm. 2. were created sequentially on the medial and lateral femoral condyles of six human cadaveric knees with intact ligaments and menisci. Chondral defects were created 1.0cm from the femoral notch posteriorly. The knees were fixed to a uniaxial load frame and loaded from 0N to 600N at full extension. Contact pressures between the femoral and tibial condyles were measured using pressure mapping sensors. The peak contact pressure was defined as the highest value in the 2.54mm. 2. area around the defect. The location of the peak contact pressure was determined relative to the centre of the defect. Peak contact pressure was significantly different between (4.30MPa) 0.09cm. 2. and (6.91MPa) 1.0cm. 2. defects (p=0.04) on the medial condyle. On the lateral condyle, post-hoc analysis showed differences in contact pressures between (3.63MPa) 0.09cm. 2. and (5.81MPa) 1.0cm. 2. defect sizes (p=0.02). The location of the stress point shifted from being posteromedial (67% of knees) to anterolateral (83%) after reaching a 0.49cm. 2. defect size (p < 0.01) in the medial condyle. Conversely, the location of the peak contact pressure point moved from being anterolateral (50%) to a posterolateral (67%) location in defect sizes greater than 0.49cm. 2. (p < 0.01). Changes in contact area redistribution and cartilage stress from 0.49cm. 2. to 1.0cm. 2. impact adjacent cartilage integrity. The location of the maximum stress point also varied with larger defects. This study suggests that size cutoffs exist earlier in the natural history of chondral defects, as small as 0.49cm. 2. , than previously studied, suggesting a lower threshold for intervention

Stand-alone anterior lumbar interbody fusion (ALIF) provides the opportunity to avoid supplemental posterior fixation. This may reduce morbidity and complication rate, which is of special interest in patients with reduced bone mineral density (BMD). This study aims to assess immediate biomechanical stability and radiographic outcome of a stand-alone ALIF device with integrated screws in specimens of low BMD. Eight human cadaveric spines (L4-sacrum) were instrumented with SynFix-LR™ (DePuy Synthes) at L5/S1. Quantitative computed tomography was used to measure BMD of L5 in AMIRA. Threshold values proposed by the American Society of Radiology 80 and 120 mg CaHa/mL were used to differentiate between Osteoporosis, Osteopenia, and normal BMD. Segmental lordosis, anterior and posterior disc height were analysed on pre- and postoperative radiographs (Fig 1). Specimens were tested intact and following instrumentation using a flexibility protocol consisting of three loading cycles to ±7.5 Nm in flexion-extension, lateral bending, and axial rotation. The ranges of motion (ROM) of the index level were assessed using an optoelectronic system. BMD ranged 58–181mg CaHA/mL. Comparison of pre- and postoperative radiographs revealed significant increase of L5/S1 segmental lordosis (mean 14.6°, SD 5.1, p < 0.001) and anterior disc height (mean 5.8mm, SD 1.8, p < 0.001), but not posterior disc height. ROM of 6 specimens was reduced compared to the intact state. Two specimens showed destructive failure in extension. Mean decrease was most distinct in axial rotation up to 83% followed by flexion-extension. ALIF device with integrated screws at L5/S1 significantly increases segmental lordosis and anterior disc height without correlation to BMD. Primary stability in the immediate postoperative situation is mostly warranted in axial rotation. The risk of failure might be increased in extension for some patients with reduced lumbar BMD, therefore additional posterior stabilization could be considered. For any figures or tables, please contact the authors directly

Decellularization techniques have advanced to reduce the risk of immune rejection in transplantation. Validation of these protocols typically relies on Crapo's criteria. 1. , which include the absence of visible nuclei and low DNA content. In our study, five decellularization protocols were compared to determine the optimal approach for human fascia lata (HFL) samples. However, our findings raised questions as to why recipients can still develop immunity despite meeting validation criteria. HFL samples were decellularized using four protocols with SDS-Triton X100-DNase (D1 to D4-HFL) and one protocol using solvent-detergent-based baths (D5-HFL). The decellularized samples (D-HFL) were compared to native samples (N-HFL) using histology, and DNA content was measured. The human leukocyte antigen (HLA) content within the matrix was assessed using western blot analysis. Both D-HFL and N-HFL samples, along with negative control patches, were implanted in the backs of 28 Wistar rats. Anti-human IgG serum levels were evaluated after one month. H&E and Hoechst staining revealed the absence of residual cells in all decellularization protocols. DNA content was consistently below the critical threshold (p<0.05). All implanted D-HFL samples resulted in significantly lower anti-human IgG levels compared to N-HFL (p<0.01). However, 2.5 out of 4 rats developed immunity after being implanted with D1 to D4-HFL, with varying levels of anti-human IgG. Only rats implanted with D5-HFL showed undetectable levels of IgG and were considered non-immunized. Western blot analysis indicated that only D5-HFL had a residual HLA content below 1%. The literature on decellularization has primarily relied on Crapo's criteria, which do not consider the role of HLA mismatch in acute immune rejection. Our results suggest that a residual HLA content below 1% should also be considered to prevent immunization, even if other validation criteria are met. Further research is needed to evaluate the impact of residual HLA levels on human allotransplantation outcomes

Abstract. Background. Ultrasonic cutting of bone boasts many advantages over alternatively powered surgical instruments, including but not limited to: elimination of swarf, reduced reaction forces, increased precision in cutting and reduced adjacent soft tissue damage, reduced post-operative complications such as bleeding and bone fracture, reduced healing time, reduced intra-operative noise and ease of handling. Despite ultrasonic cutting devices being well established in oral and maxillofacial surgery, applications in orthopaedic surgery are more niche and are not as well understood. The aim of this study was to investigate the cutting speed (mm/s) and cutting forces (N) of orthopaedic surgeons using a custom-designed state of the art ultrasonic cutting tool to cut fresh human bone samples. Methods. A setup based on the Robot Operating System (ROS) and AprilTag was designed to track and to record the real time position of the ultrasonic cutting tool in space. Synchronised load cell axial force readings of three separate orthopaedic surgeons during ultrasonic cutting were recorded. Each surgeon was asked to find a comfortable position that reflects as close as possible their clinical handling of a cutting instrument used in surgery, and to perform two cuts in each of three samples of human cortical bone. Bone samples were obtained following ethical approval from an institutional review board (ethics approval number: SR1342) and prior informed consent was obtained from all patients. Bone samples were extracted from the femoral neck region of three hip osteoarthritis patients. During cutting, surgeons were allowed a total cutting time of one minute and cutting was conducted using an ultrasonic tool with frequency of a 35kHz (35.7 µm peak to peak displacement amplitude) under constant irrigation using a MINIPULS® 3 Peristaltic pump (38 revolutions per minute) using Phosphate-Buffered Saline (PBS) at 25°C. From the recorded data, the average instantaneous cutting velocity was calculated and the maximum cutting force was identified. Results. All surgeons assumed a back-and-forth cutting motion, variation in the applied cutting force was observed. The average vertical cutting speed, axial cutting force and cutting depth across all surgeons and all samples was 1.64 mm/s, 1.91 N and 0.73 mm, respectively. While increasing the axial cutting force resulted in a deeper cut, overloading of the ultrasound transducer occurred when the tool advanced too quickly into the bone tissue during cutting. The exact force threshold, or the optimal speed at which the surgeon can maintain a constant force during cutting, requires further investigation. Conclusions. In this study, all surgeons cut using a back-and-forth cutting motion, with variation in the applied cutting force which may ultimately inform which clinical applications in orthopaedic engineering are most suitable for this technology. Applying too much force caused overloading of the ultrasound transducer, which is a limitation with the current cutting tool. The results from this study may facilitate the eventual uptake of ultrasonic cutting tools for application in orthopaedic surgery. Declaration of Interest. (b) declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported:I declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project

Aim. To investigate the validity of threshold values for the Oxford Hip and Knee Score (OHS and OKS) for treatment success 12 months after total knee or hip replacement. Methods. Questionnaires were administered to patients undergoing total hip (THA) or knee (TKA) replacement before and 12 months after surgery alongside questions assessing key accepted aspects of treatment success (satisfaction, pain relief, functional improvement) to form a composite criterion of success and assessed using receiver operator characteristic (ROC) analysis. Thresholds providing maximum sensitivity and specificity for predicting treatment success were determined for the total sample and subgroups defined by pre-surgery scores. Results. Data was available for 3203 THA and 2742 TKA patients. Applying the composite treatment success criterion, 52.7% of the TKA and 65.6% of the THA sample reported a successful treatment outcome. Accuracy for predicting treatment success was high for the OHS and OKS (both areas under curve 0.87). For the OHS a threshold value of 37.5 points showed highest sensitivity and specificity in the total sample, while for the OKS the optimal threshold was 32.5 points. Depending on pre-surgery scores optimal thresholds varied between 32.5 and 38.5 for the OHS and 28.5 and 35.5 for the OKS. Conclusions. This is the first study to apply a comprehensive composite “success” anchor to the OHS and OKS in order to determine thresholds for successful treatment with total joint replacement. Compared to widely publicised postoperative satisfaction metrics, far fewer patients report success with our composite anchor

Ligaments and tendons are connective tissues with a highly hierarchical structure, from collagen fibres, to fibrils and fascicules. Their intricate structural arrangement produces an anisotropic non-linear elastic mechanical behaviour and a complex damage pattern before failure. Recent constitutive models have been developed with all parameters describing the structure of the tissue, with the advantage that they can in theory be measured on the tissue rather than being phenomenologically-derived. This is an ideal framework to model damage as its onset and propagation can be associated to changes in the structure directly. In this preliminary study, the possibility to identify damage mechanisms in the tissue structure using in silico models was analysed for both the anterior cruciate ligament, with fascicules forming a helix with its longitudinal axis, and the patellar tendon, with fascicules co-aligned with its longitudinal axis. Tissues of interest were modelled as cylinders submitted to uniaxial tension. Damage was modelled as either a reduction of collagen volume fraction with increased strain, assuming the number of collagen fibres sustaining load decreases as fibres fail, or a reduction of the modulus of the fibres, assuming pre-failure damage of the fibres. Each damage mechanism was associated with a damage variable with different fibre stretch threshold for damage initiation and assuming linear variation of damage until an arbitrary failure point. The apparent behaviour of the modelled tissues was significantly different as damage thresholds, damage mechanisms, type of fascicules were varied. This preliminary work showed that using a structural constitutive model to describe occurrence and propagation of structural damage in an in silico model of hierarchical connective tissues is a framework that can clearly differentiate at a macroscopic level between different values of damage threshold and different damage mechanisms for tissue with co-aligned or helical fascicules