Objectives

The aim of the current study was to assess whether calcaneal broadband ultrasound attenuation (BUA) can predict whole body and regional dual-energy x-ray absorptiometry (DXA)-derived bone mass in healthy, Australian children and adolescents at different stages of maturity.

Abstract. Objective. This study assesses the prevalence of major and minor discordance between hip and spine T scores using Radiofrequency Echographic Multi-spectrometry (REMS). REMS is a novel technology that uses ultrasound and radiofrequency analysis to measure bone density and bone fragility at the hip and lumbar spine. The objective was to compare the results with the existing literature on Dual-Energy X-ray Absorptiometry (DEXA) the current “gold standard” for bone densitometry. REMS and DEXA have been shown to have similar diagnostic accuracy, however, REMS has less human input when carrying out the scan, therefore the rates of discordance might be expected to be lower than for DEXA. Discordance poses a risk of misclassification of patients’ bone health status, causing diagnostic ambiguity and potentially sub-optimal management decisions. Reduction of discordance rates therefore has the potential to significantly improve treatment and patient outcomes. Methods. Results from 1,855 patients who underwent REMS investigations between 2018 and 2022 were available. Minor discordance is defined as a difference of one World Health Organisation (WHO) diagnostic classification (Normal / Osteopenia or Osteopenia / Osteoporosis). Major discordance is defined as a difference of two WHO diagnostic classifications (Normal / Osteoporosis). The results were compared with reported DEXA discordance rates. Results. 1,732 individuals had both hip and spine T scores available for analysis. There were 267 cases of discordance. No instances of major discordance were observed. The minor discordance rate was 15.4%. 6.5% of the REMS scans with minor discordance showed > 1.0 standard deviation (SD) difference between the T scores of the hip and spine. 19.4% had differences of between 0.6 SD and 1.0 SD while 73.9% had ≤ 0.5 SD or less. In 24.5% of the cases of REMS discordance the hip T scores were greater than the spine and in 75.5% of cases the spine T score was greater than the hip. Conclusions. The current analysis is the largest of its kind. It demonstrates that REMS has an overall lower rate of discordance than reported DEXA rates. Major discordance rates with DEXA range from 2–17%, but REMS avoids many of the positioning problems and post-processing errors inherent in DEXA scanning, which might account for the absence of major discordance. Rates of minor discordance in DEXA scans range between 38–51%. The REMS minor discordance rate being much lower than these rates suggests that it has the potential to enhance diagnostic accuracy considerably. Most REMS discordance results showed ≤ 0.5 SD variance between the T scores of the two sites, indicating close correlation in the bone densitometry analysis. Most studies of DEXA discordant results confirm that spinal T scores are more often higher than at the hip. The REMS results concur with this observation. Considering the comparable accuracy rates that have been shown between REMS and DEXA, with its much lower discordance rate, REMS can potentially improve current medical practice and enhance patient care. 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

Sarcopenia is a progressive and generalized skeletal muscle disorder that involves loss of muscle mass and function. It is associated with increased adverse outcomes including falls, functional decline, frailty and mortality and affects 65% of people over the age of 65 more than half of people aged 80 and above. The factors that cause and worsen sarcopenia are categorised into two groups. The primary aetiological factor is ageing and the secondary factors include disease, physical inactivity, and poor nutrition. Sarcopenia is considered to be ‘primary' (or age-related) when no other specific cause is evident. However, a number of ‘secondary' factors may be present in addition to ageing. Sarcopenia can occur secondary to a systemic or inflammatory disease, including malignancy and organ failure. Physical inactivity is one of the major contributors to the development of sarcopenia, whether due to a sedentary lifestyle or to disease related immobility or disability. Furthermore, sarcopenia can develop as a result of inadequate protein consumption. Biomarkers are objective and quantifiable characteristics of physiological and pathophysiological processes. Biomarkers can be used to predict the development of sarcopenia in older susceptible adults and enable early interventions that can reduce the risk of physical disability, the co-morbidities associated with the loss of muscle mass and the poor health outcomes that result from sarcopenia. Non-invasive imaging technologies can be used as biomarkers to detect loss of skeletal muscle mass in sarcopenia include bone densitometry, computed tomography, ultrasound and magnetic resonance imaging. However, imaging requires sophisticated and expensive equipment that is not available in a resource poor setting. Therefore, markers of skeletal muscle strength and fitness and soluble biochemical markers in blood may be used as alternative biomarkers. Studies on sarcopenia have identified numerous soluble biochemical biomarkers. These biomarkers can be divided into two groups: “muscle-specific” and “non-muscle-specific” biomarkers. Since sarcopenia is associated with rapid skeletal muscle wasting, the skeletal muscle-specific isoform of troponin T may be considerate a useful biomarker of sarcopenia, since high troponin levels in blood are an expression of muscle wasting. Peptides derived from collagen type VI turnover may be potential biomarkers of sarcopenia. We have recently conducted a systematic review to summarize the data from recent mass-spectrometry based proteomic studies of the secretome of skeletal muscle cells in response to disease, exercise or metabolic stress in order to identify the proteins involved in muscle breakdown. Developing robust in vitro models for the study of sarcopenia using primary muscle cells is a high priority as is exploiting the in vitro models to understand catabolic and inflammatory processes and molecular mechanisms involved in sarcopenia. Co-cultures with adipose-derived and other cells may be used to screen for small molecules and biologicals capable of inhibiting the catabolic and inflammatory pathways involved in sarcopenia. This presentation reviews recent progress in this area and outlines opportunities for future research on sarcopenia

Proximal femur fractures are common in the elderly population. The aim of this study was to determine the relationship between fracture type and proximal femoral geometric parameters. We retrospectively studied the electronic medical records of 85 elderly patients over 60 years of age who were admitted to the orthopedic department with hip fractures between January 2016 and January 2018 in a training and research hospital in Turkey. Age, fracture site, gender, implant type and proximal femoral geometry parameters (neck shaft angle [NSA], center edge angle [CEA], femoral head diameter [FHD], femoral neck diameter [FND], femoral neck axial length [FNAL], hip axial length [HAL], and femoral shaft diameter [FSD]) were recorded. Patients with femoral neck fractures and femur intertrochanteric fractures were divided into two groups. The relationship between proximal femoral geometric parameters and fracture types was examined. SPSS 25.0 (IBM Corparation, Armonk, New York, United States) program was used to analyze the variables. Independent samples t test was used to compare the fracture types according to NSA, FHD, FND and FSD variables. A statistically significant difference was found in FSD (p=0,002) and age (p=0,019). FSD and age were found to be greater in intertrochanteric fractures than neck fractures. Gender, site, CEA, FNAL, HAL, NSA, FHD and FND parametres were not significantly different. In the literature, it is seen that different results have been reached in different studies. In a study conducted in the Chinese population, a significant difference was found between the two groups in NSA, CEA and FNAL measurements. In a study conducted in the Korean population, a significant difference was found only in NSA measurements. The FSD is generally associated with bone mineral densitometry in the literature and has been shown to be a risk factor for fracture formation. However, a study showing that there is a relationship between FSD and fracture type is not available in the literature. In this study; FSD was found to be higher in intertrochanteric fractures (p = 0.002). However, for the clinical significance of this difference, we think that larger patient series and biomechanical studies are needed

Tenocytes from several mammal species have been shown to be prone to phenotypic drift at early sub-culture passages. In the present study we compared allogenic and xenogenic serum supplementation suitability as a supplement for the in vitro expansion of equine tenocytes (eTCs), in combination with the presence or absence of crowding conditions. eTCs were isolated from superficial digital flexor tendon and expanded in normal growth medium containing DMEM, 10% appropriate serum, 1% penicillin/streptomycin solution. Isolation was performed by migration method in growth medium containing the selected serum. Silver staining, densitometry, zymography, immunofluorescence, metabolic activity, proliferation, viability and morphology were performed after 3, 5 and 7 days in culture with a seeding density of 10,000 cells/cm2. Treatment conditions were equine serum (ES) or foetal bovine serum (FBS), with or without 75 μg/mL of crowding agent carrageenan (CR). Viability and metabolic activity of eTCs were affected by FBS. eTCs in ES reached higher cell density than in FBS in day 7, especially with CR. Morphology of eTCs was maintained under different sera. Silver staining on pepsin digested cell layers shows that collagen type I deposition rate is remarkably enhanced in the presence of CR in all conditions. Immunofluorescence showed increased expression for collagen I, III, V and VI in both sera in the presence of CR. Deposition of all collagen types but type VI was increased by ES supplementation. We conclude that ES in combination with CR can represent a reliable choice for the ex vivo expansion of eTCs

Introduction. Precision error (PE) in Dual Energy X-Ray Absorptiometry (DXA) is important for accurate monitoring of changes in Bone-Mineral-Density (BMD). It has been demonstrated that BMD PE increases with increasing BMI. In vivo PE for the Trabecular-Bone-Score (TBS) has not been reported. This study aimed to evaluate the short-term PE (STPE)) of BMD and TBS and to investigate the effect of obesity on DXA PE. Method. DXA lumbar spine scans (L1–L4) were performed using GE Lunar Prodigy. STPE was measured in 91 women (Group A) at a single visit by duplicating scans with repositioning in-between. PE was calculated as the percentage coefficient of variation (%CV). Group A was sub-divided into four groups based on BMI (A.1. <25kg/m2, A.2. 25–29.9kg/m2, A.3. 30–35kg/m2 and A.4. >35kg/m2) to assess the effect of obesity on STPE. Abnormally different vertebrae were excluded from the analysis in accordance with The International Society for Clinical Densitometry (ISCD) recommendations. Results. The Group A STPE was 1.26 % for BMD and 2.04% for TBS. Short-term PE for BMD and TBS respectively in the BMI subgroups was: A.1. 1.07% and 1.82%, A.2. 1.34% and 2.26%, A.3. 1.25% and 2.35%, A.4. 1.68% and 1.82%. Conclusion. The results show that STPE is higher for TBS than for BMD. Short-term PE for both BMD and TBS are adversely affected by increasing BMI but this effect is mitigated in the highest BMI category where use of the ‘thick’ scanning mode improves signal to noise ratio

Infection of human cartilage with HIV in vivo has not previously been reported. Specimens of articular cartilage taken at postmortem from ten patients who were HIV-positive were examined. Two had AIDS and eight were believed to have stage-2 disease. The standard polymerase chain reaction (PCR) protocol was modified to allow semiquantitative analysis of the samples. Oligonucleotide primers labelled with . 32. P gamma-ATP were used to detect a segment of HIV DNA and a control DNA gene segment (HLA genome) to estimate the ratio of infected cells. The . 32. P-labelled PCR products were separated on acrylamide gels and visualised directly by autoradiography and computer densitometry. Infection of human cartilage in vivo was demonstrated in nine of the ten samples in which the PCR analysis was positive. The other did not react sufficiently to produce detectable radiolabelled PCR product despite repeated DNA digestion and extraction. Cartilage infected with HIV could be a potential source of HIV when used in operations

We investigated several factors which affect the stability of cortical screws in osteoporotic bone using 18 femora from cadavers of women aged between 45 and 96 years (mean 76). We performed bone densitometry to measure the bone mineral density of the cortical and cancellous bone of the shaft and head of the femur, respectively. The thickness and overall bone mass of the cortical layer of the shaft of the femur were measured using a microCT scanner. The force required to pull-out a 3.5 mm titanium cortical bone screw was determined after standardised insertion into specimens of the cortex of the femoral shaft. A significant correlation was found between the pull-out strength and the overall bone mass of the cortical layer (r. 2. = 0.867, p < 0.01) and also between its thickness (r. 2. = 0.826, p < 0.01) and bone mineral density (r. 2. = 0.861, p < 0.01). There was no statistically significant correlation between the age of the donor and the pull-out force (p = 0.246), the cortical thickness (p = 0.199), the bone mineral density (p = 0.697) or the level of osteoporosis (p = 0.378). We conclude that the overall bone mass, the thickness and the bone mineral density of the cortical layer, are the main factors which affect the stability of a screw in human female osteoporotic cortical bone

Summary. In a rabbit model of early osteoarthritis, structural changes in femoral condyle cartilage were severer in the lateral compartment and preceded alterations in the underlying bone. In the medial compartment, altered bone properties occurred together with structural changes in cartilage. Introduction. Early osteoarthritic changes in cartilage have been previously studied through anterior cruciate ligament transection (ACLT) in rabbits. However, parallel changes in the structure of subchondral and trabecular bone at 4 weeks after ACLT are not known. Methods. Skeletally mature 14-month old New Zealand white rabbits (n=8) underwent ACLT in the left knee, while right knees were used as controls (CTRL). Femoral condyles (FCs) were harvested at 4 weeks after ACLT. INDENTATION TESTING. Stepwise stress-relaxation tests were performed on medial and lateral FC cartilage (100%/s ramp rate, 3×5% step, 15 min relaxation time). Sinusoidal loading was then applied (amplitude 4% of thickness, 1Hz, 4 cycles). Equilibrium (Eeq) and dynamic (Ed) moduli were derived from stress-relaxation and sinusoidal tests, respectively. STRUCTURAL ANALYSIS OF CARTILAGE. Polarised light microscopy (PLM) and digital densitometry (DD) were used to analyze the collagen orientation angle (COA) and proteoglycan content in the cartilage samples. STRUCTURAL ANALYSIS OF BONE. Distal compartments of FCs were scanned using a high-resolution µCT scanner (Skyscan 1172, Belgium) with an isotropic voxel size of 25 µm. µCT data were imported into Mimics (Materialise, Belgium) for segmentation. 2×2×4 mm. 3. volumes of interest (VOIs) were placed in weight-bearing regions of medial and lateral FCs. Subchondral bone plate thickness (Pt.Th), trabecular volume fraction (BV/TV), trabecular thickness (Tb.Th), structural model index (SMI) and trabecular separation (Tb.Sp) were calculated using the CTAnalyzer software (Skyscan) from the VOIs. STATISTICAL TESTS. Mixed linear model for cartilage parameters and Wilcoxon signed-rank test for bone parameters were used to compare ACLT and CTRL groups (p < 0.05). Results. In both lateral and medial FC compartments, Eeq was significantly smaller in ACLT than in CTRL cartilage. In the medial compartment, also Ed was significantly smaller in ACLT than in CTRL cartilage. As a result of ACLT, significant alterations in the COA extended deeper into cartilage in the lateral than medial compartment, while proteoglycan content was reduced significantly and similarly in both lateral and medial FC cartilages. After ACLT, Pt.Th was significantly reduced in the medial compartment, while no changes were observed in the lateral compartment. Furthermore, only in the medial compartment, both BV/TV and Tb.Th were significantly smaller in the ACLT compared to the CTRL group. Discussion. The study showed that disruption of the collagen architecture in the ACLT joint cartilage extended into the middle zone only in the lateral FC compartment. Instead, thinning of the subchondral bone plate combined with resorption of trabecular bone was observed only in the medial FC compartment. The former finding reflects early osteoarthritic changes, while the latter finding may be indicative of a diminished loading in the medial FC compartment, as caused by ACLT

Introduction. The use of platelet-rich concentrate (PRC) to enhance the healing response in tendon repair is currently an area of considerable interest. Activated platelets release a cocktail of growth factors and ECM regulating molecules. Previous work suggests that tenocytes are activated by contact with these clot-derived molecules. Our studies on tenocytes and PRC aim to establish the direct molecular and functional effects of PRC on tenocytes and to support the clinical research on Achilles tendon repair taking place within our group. We hypothesise that applying PRC to human tenocytes in culture will increase proliferation rate and survival by activating relevant signalling pathways. Materials and Methods. Using a centrifugation method, PRC was extracted from fresh human whole blood. The PRC was immediately clotted and left in medium overnight to release biological factors (at least 95% of presynthesized growth factors are secreted in the first hour of activation). 1. Human tenocytes derived from explanted healthy hamstring were used for up to three passages. Cells were treated with varying concentrations of PRC-conditioned medium and assessed for viable cell number (Alamar Blue™ fluorescence) and proliferation (Ziva™ Ultrasensitive BrdU assay) after 72hrs. For western blotting, cells were treated with 10% PRC for 5 or 30 minutes. Antibodies to P-ERK and P-Akt detected the active protein state on the blot, followed by membrane stripping and re-probing with pan antibodies. Quantification was achieved by densitometry using Visionworks software v. 6.7.1. Results. PRC-conditioned medium affected tenocytes in a dose-dependent manner. Viable number of tenocytes was significantly increased by 10% PRC-conditioned medium compared to controls (One-way ANOVA, Tukey's post-hoc test P<0.001) after 72hrs. 10% PRC-conditioned medium also demonstrated time-dependence with viable tenocyte number significantly increasing between 24 and 72hrs (One-way ANOVA, Bonferroni's post-hoc test P<0.001). After 72hrs, tenocyte proliferation significantly increased in the presence of 5% and 10% PRC-conditioned media compared to controls (One-way ANOVA, Tukey's post-hoc test P<0.05 and P<0.001 respectively). ERK and Akt phosphorylation was strongly stimulated by treatment with 10% PRC-conditioned medium for 5 minutes compared to controls, and remained high after a 30 minute application time. Discussion and Conclusions. Factors released by activated PRC act upon human tendon cells to strongly increase viable cell number and proliferation, which would, in vivo, directly support the healing response, independent of any additional beneficial effects on vascular repair. Both ERK and Akt are pivotal kinases in signalling pathways that favour survival and proliferation. It is clear that both signalling pathways are immediately and strongly activated by PRC, suggesting a clear benefit via both stimulated cell cycle and cell survival in the environmentally compromised conditions of a healing ruptured tendon. This conclusion is strongly supported by previous work on platelet releasates and ERK signalling in other cell types

Objectives

The treatment of osteoporotic fractures is a major challenge, and the enhancement of healing is critical as a major goal in modern fracture management. Most osteoporotic fractures occur at the metaphyseal bone region but few models exist and the healing is still poorly understood. A systematic review was conducted to identify and analyse the appropriateness of current osteoporotic metaphyseal fracture animal models.

This study was designed to test the hypothesis that the sensory innervation of bone might play an important role in sensing and responding to low-intensity pulsed ultrasound and explain its effect in promoting fracture healing. In 112 rats a standardised mid-shaft tibial fracture was created, supported with an intramedullary needle and divided into four groups of 28. These either had a sciatic neurectomy or a patellar tendon resection as control, and received the ultrasound or not as a sham treatment. Fracture union, callus mineralisation and remodelling were assessed using plain radiography, peripheral quantitative computed tomography and histomorphology.

Daily ultrasound treatment significantly increased the rate of union and the volumetric bone mineral density in the fracture callus in the neurally intact rats (p = 0.025), but this stimulating effect was absent in the rats with sciatic neurectomy. Histomorphology demonstrated faster maturation of the callus in the group treated with ultrasound when compared with the control group. The results supported the hypothesis that intact innervation plays an important role in allowing low-intensity pulsed ultrasound to promote fracture healing.

Medial open-wedge high tibial osteotomy has been gaining popularity in recent years, but adequate supporting material is required in the osteotomy gap for early weight-bearing and rapid union. The purpose of this study was to investigate whether the implantation of a polycaprolactone-tricalcium phosphate composite scaffold wedge would enhance healing of the osteotomy in a micro pig model. We carried out open-wedge high tibial osteotomies in 12 micro pigs aged from 12 to 16 months. A scaffold wedge was inserted into six of the osteotomies while the other six were left open. Bone healing was evaluated after three and six months using plain radiographs, CT scans, measurement of the bone mineral density and histological examination.

Complete bone union was obtained at six months in both groups. There was no collapse at the osteotomy site, loss of correction or failure of fixation in either group. Staining with haematoxylin and eosin demonstrated that there was infiltration of new bone tissue into the macropores and along the periphery of the implanted scaffold in the scaffold group. The CT scans and measurement of the bone mineral density showed that at six months specimens in the scaffold group had a higher bone mineral density than in the control group, although the implantation of the polycaprolactone-tricalcium phosphate composite scaffold wedge did not enhance healing of the osteotomy.

We performed a biomechanical study to compare the augmentation of isolated fractured vertebral bodies using two different bone tamps. Compression fractures were created in 21 vertebral bodies harvested from red deer after determining their initial strength and stiffness, which was then assessed after standardised bipedicular vertebral augmentation using a balloon or an expandable polymer bone tamp.

The median strength and stiffness of the balloon bone tamp group was 6.71 kN (sd 2.71) and 1.885 kN/mm (sd 0.340), respectively, versus 7.36 kN (sd 3.43) and 1.882 kN/mm (sd 0.868) in the polymer bone tamp group. The strength and stiffness tended to be greater in the polymer bone tamp group than in the balloon bone tamp group, but this difference was not statistically significant (strength p > 0.8, and stiffness p = 0.4).