Three-dimensional image registration improves the long-term precision of in vivo micro-computed tomographic measurements in anabolic and catabolic mouse models.

Micro-computed tomography (micro-CT) is a widely used technique to track bone structural and mineral changes in small animals in vivo. Precise definition of volumes of interest (VOIs) in follow-up scans is required to accurately quantify these changes. To improve precision, VOIs can be transferred from baseline images onto follow-ups using image registration. We studied the performance of a registration procedure applied to in vivo data sets of anabolic and osteoporotic bone changes in mice. Micro-CT image data from two separate CD1 mouse data sets were studied. The first included a group treated with parathyroid hormone (PTH) and control and the second, an ovariectomy (OVX) group and control. Micro-CT was performed once per week for 4 weeks at the proximal tibia starting at treatment onset (PTH data set) or after surgery (OVX data set). A series consisting entirely of user-defined VOIs and a registered series where VOIs defined at baseline were transferred to follow-ups were created. Standard bone structural and mineral measurements were calculated. Image registration resulted in a 13-56 % reduction in precision error. Significant effects of registration to detect PTH-induced changes in BV/TV and trabecular BMD were observed. When changes were very pronounced or small, the qualitative improvement observed for the registered data set did not reach statistical significance. This study documents an increase in long-term precision of micro-CT measurements with image registration. Sensitivity to detect changes was improved but not uniform for all parameters. Future study of this technique on images with a smaller voxel size (<19 µm) may capture the effect in greater detail, in particular for trabecular thickness, where changes may be too small to be observed with the voxel size used here. Our results document the value of registration and indicate that the magnitude of improvement depends on the model and treatment chosen.

Assignment of the biological value of solid breast masses based on quantitative evaluations of spectral CT examinations using electron density mapping, Zeffective mapping and iodine mapping.

OBJECTIVE: We aimed to asses, in a clinical setting, whether the newly available quantitative evaluation of electron density (ED) in spectral CT examinations of the breast provide information on the biological identity of solid breast masses and whether ED maps yield added value to the diagnostic information of iodine maps and Zeff maps calculated from the same CT image datasets. METHODS: All patients at the University Breast Cancer Center who underwent a clinically indicated Dual Layer Computed Tomography (DLCT) examination for staging of invasive breast cancer from 2018 to 2020 were prospectively included. Iodine concentration maps, Zeff maps and ED maps were automatically reconstructed from the DLCT datasets. Region of interest (ROI) based evaluations in the breast target lesions and in the aorta were performed semi-automatically in identical anatomical positions using dedicated evaluation software. Case-by-case evaluations were carried independently by 2 of 4 radiologists for each examination, respectively. Statistical analysis derived from the ROIs was done by calculating ROC/AUC curves and Youden indices. RESULTS: The evaluations comprised 166 DLCT examinations. In the ED maps the measurements in the breast target lesions yielded Youden cutpoints of 104.0% (reader 1) and 103.8% (reader 2) resulting in AUCs of 0.63 and 0.67 at the empirical cutpoints. The variables "Zeff" and "iodine content" derived from the target lesions showed superior diagnostical results, with a Youden cutpoint of 8.0 mg/ml in the iodine maps and cutpoints of 1.1/1.2 in the Zeff maps the AUCs ranging from 0.84 to 0.85 (p = 0.023 to <0.000). The computational combination of Zeff and ED measurements in the target lesions yielded a slight AUC increase (readers 1: 0.85-0.87; readers 2: 0.84-0.94). The ratios of the measured values in the target lesions normalized to the values measured in the aorta showed comparable results. The AUCs of ED derived from the cutpoints showed inferior results to those derived from the Zeff maps and iodine maps (ED: 0.64 and 0.66 for reader 1 and 2; Zeff: 0.86 for both readers; iodine content: 0.89 and 0.86 for reader 1 and 2, respectively). The computational combination of the ED results and the Zeff measurements did not lead to a clinically relevant diagnostic gain with AUCs ranging from 0.86 to 0.88. CONCLUSIONS: Quantitative assessments of Zeff, iodine content and ED all targeting the physical and chemical aspects of iodine uptake in solid breast masses confirmed diagnostically robust cutpoints for the differentiation of benign and malignant findings (Zeff < 7.7, iodine content of <0.8 mg/ml). The evaluations of the ED did not indicate any added diagnostic value beyond the quantitative assessments of Zeff and iodine content. Further research is warranted to develop suitable clinical indications for the use of ED maps.

Detecting Bone Marrow Edema of the Extremities on Spectral Computed Tomography Using a Three-Material Decomposition.

BACKGROUND: Detecting bone marrow edema (BME) as a sign of acute fractures is challenging on conventional computed tomography (CT). This study evaluated the diagnostic performance of a three-material decomposition (TMD) approach for detecting traumatic BME of the extremities on spectral computed tomography (SCT). METHODS: This retrospective diagnostic study included 81 bone compartments with and 80 without BME. A TMD application to visualize BME was developed in collaboration with Philips Healthcare. The following bone compartments were included: distal radius, proximal femur, proximal tibia, distal tibia and fibula, and long bone diaphysis. Two blinded radiologists reviewed each case independently in random order for the presence or absence of BME. RESULTS: The interrater reliability was 0.84 (p < 0.001). The different bone compartments showed sensitivities of 86.7% to 93.8%, specificities of 84.2% to 94.1%, positive predictive values of 82.4% to 94.7%, negative predictive values of 87.5% to 93.3%, and area under the curve (AUC) values of 85.7% to 93.1%. The distal radius showed the highest sensitivity and the proximal femur showed the lowest sensitivity, while the proximal femur presented the highest specificity and the distal tibia presented the lowest specificity. CONCLUSIONS: Our TMD approach provides high diagnostic performance for detecting BME of the extremities. Therefore, this approach could be used routinely in the emergency setting.

Intermodal correlation of quantitative CT-data and MRI-biomarkers derived from synchronous spectral CT-maps and breast MRI-examinations with molecular biomarkers in invasive ductal breast carcinomas.

OBJECTIVE: To asses the correlation of data derived from dual-layer (DL)-CT material-maps and breast MRI data with molecular biomarkers in invasive breast carcinomas. METHODS: All patients at the University Breast Cancer Center who underwent a clinically indicated DLCT-scan and a breast MRI for staging of invasive ductal breast cancer from 2016 to 2020 were prospectively included. Iodine concentration-maps, and Zeffective-maps were reconstructed from the CT-datasets. T1w- and T2w-signal intensities, ADC and the clustered shapes of the dynamic-curves (washout, plateau, persistent) were derived from the MRI-datasets. ROI-based evaluations of the cancers and the reference "musculature" were performed semi-automatically in identical anatomical positions using dedicated evaluation software. Statistical analysis was essentially descriptive using Spearmans rank correlation and (multivariable) partial correlation. RESULTS: The signal intensities measured in the 3rd phase of the contrast dynamics correlated at an intermediate level of significance with the iodine content and the Zeffective-values derived from the breast target lesions (Spearmans rank correlation-coefficient r = 0.237/0.236, p = 0.002/0.003). The bivariate and the multivariate analyses displayed correlations of an intermediate significance level of the iodine content and the Zeff-values measured in the breast target lesions with immunhistochemical subtyping (r = 0.211-0.243, p = 0.002-0.009, respectively). The Zeff-values showed the strongest correlations when normalized to the values measured in the musculature and in the aorta (r = -0.237 to -0.305, r=<0.001-0.003). The MRI-assessments showed correlations of intermediate to high significance and low to intermediate significance between the ratios of the T2w-signal intensities and the trends of the dynamic curves measured in the breast target lesions and in musculature and immunohistochemical cancer subtyping, respectively (T2w: r = 0.232-0.249, p = 0.003/0.002; dynamics: r = -0.322/-0.245, p=<0.001/0.002). The ratios of the clustered trends of the dynamic curves measured in the breast target lesions and in musculature correlated with tumor grading on intermediate significance level (r = -0.213 and -0.194, p = 0.007/0.016) and with Ki-67 on a low significance level (bivariate analysis: r = -0.160, p = 0.040). There was only a weak correlation between the ADC-values measured in the breast target lesions and HER2-expression (bivariate ansalysis: r = 0.191, p = 0.030). CONCLUSIONS: Our preliminary results indicate that evaluation of perfusion based DLCT-data and MRI-biomarkers show correlations with the immunhistochemical subtyping of invasive ductal breast carcinomas. Further clinical research is warranted in order to validate the value of the results and define clinical situations in which the use of the described DLCT-biomaker and MRI biomarkers may be helpful in clinical patient care.

Dual-Layer Spectral-Computed Tomography Enhances the Separability of Calcium-Based Implant Material from Bone: An Ex Vivo Quantitative Imaging Study.

Local treatment of bone loss with an injection of a resorbable, calcium-based implant material to replace bone has a long history of clinical use. The in vivo discrimination of changes in bone versus implant is challenging with standard computed tomography (CT). However, spectral-CT techniques enable the separation between tissues of similar densities but different chemical compositions. Dual-layer spectral-CT imaging and postprocessing analysis methods were applied to investigate the separability of AGN1 (a triphasic calcium-based implant) and bone after AGN1 injection in n = 10 male cadaveric femurs ex vivo. Using the area under the curve (AUC) from receiver-operating characteristic (ROC) analyses, the separability of AGN1 from bone was assessed for AGN1 (postoperatively) versus compact and versus femoral neck cancellous bone (both preoperatively). CT techniques included conventional Hounsfield (HU) and density-equivalent units (BMD, mg hydroxyapatite [HA]/cm3 ) and spectral-CT measures of effective atomic number (Zeff) and electron density (ED). The samples had a wide range of femoral neck BMD (55.66 to 241.71 mg HA/cm3 ). At the injection site average BMD, HU, Zeff, and ED increased from 69.5 mg HA/cm3 , 109 HU, 104.38 EDW, and 8.30 Zeff in the preoperative to 1233 mg HA/cm3 , 1741 HU, 181.27 EDW, and 13.55 Zeff in the postoperative CT scan, respectively. For compact bone at the femoral shaft the preoperative values were 1124.15 mg HA/cm3 , 1648 HU, 177 EDW, and 13.06 Zeff and were maintained postoperatively. Zeff showed substantially sharper distributions and significantly greater separability compared to ED, BMD, and HU (all p < 0.002, for both regions) with average AUCs for BMD, HU, ED, and Zeff of 0.670, 0.640, 0.645, and 0.753 for AGN1 versus compact and 0.996, 0.995, 0.994, and 0.998 for AGN1 versus femoral neck cancellous sites, respectively. Spectral-CT permits better discrimination of calcium-based implants like AGN1 from bone ex vivo. Our results warrant application of spectral-CT in patients undergoing procedures with similar implants. © 2022 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).

Correlation of CT-data derived from multiparametric dual-layer CT-maps with immunohistochemical biomarkers in invasive breast carcinomas.

OBJECTIVE: To examine the correlation of quantitative measurements from material decomposition maps calculated from dual-layer CT (DLCT)-image datasets with immunohistochemical biomarkers of invasive breast carcinomas. MATERIAL AND METHODS: All patients at the University Breast Cancer Center who underwent a clinically indicated dual-layer CT-scan for staging of invasive ductal breast carcinoma from 01/2016 to 07/2020 were prospectively included. Iodine concentration maps and maps of the effective atomic numbers (Zeffective) were reconstructed from the image datasets. ROI-based evaluations of the index tumors and predefined references tissues for normalization were performed semi-automatically in identical anatomical positions using dedicated evaluation software. Statistical analysis was essentially descriptive using Spearmans rank correlation and (multivariable) partial correlation. RESULTS: Bivariate showed statistically significant correlations of iodine contents (r = -0.154/-0.202/0.180, p = 0.039/0.006/0.015), and Zeffective-values (r = -0.158/-0.199/0.179, p = 0.034/0.007/0.016) for all 184 carcinomas and the subgroup of 168 invasive ductal carcinomas. The results were confirmed by multivariate analyses with "age", "diameter" and "ACR-grade" as possible confounders. Normalization of the measured target values with those in the aorta confirmed significant correlations of iodine content and Zeffective compared to Estrogen (r = 0.174, p = 0.019), Progesteron (r = 0.168/0.177, p = 0.024/0.017), and HER2 receptor expression (r = -0.222/-0.184, p = 0.003/0.013). All CT-parameters showed significant correlations with immunohistochemical subtyping (r = 0.191/0.192, p = 0.010). CONCLUSIONS: Our preliminary results indicate that iodine content and Zeffective-values derived from DLCT-examinations correlate with hormone receptor expression in invasive breast carcinomas. Assignments to benign entities already seam feasible in clinical routine CT-diagnostics. After further investigations iodine content and Zeffective may be translated as diagnostical and prognostical biomarkers into clinical routine in the long term.

Bone quality is partially recovered after the discontinuation of RANKL administration in rats by increased bone mass on existing trabeculae: an in vivo micro-CT study.

UNLABELLED: Bone loss and recovery in a receptor activator for nuclear factor κ B ligand (RANKL)-administered rat model was assessed. Microarchitecture, mineralization and strength deteriorated faster than ovariectomy (OVX). Recovery was dependent on the loss of trabecular elements and connections. Early recovery suggests a natural mechanism in rats to overcome excess RANKL, and may have implications for long-term bone loss. PURPOSE: To compare a model for experimental osteoporosis that induces bone loss by injecting RANKL into rats to an OVX rat model, and measure subsequent recovery of bone architecture, mineralization, and mechanics after stopping injections. METHODS: Mature, healthy, female Wistar rats were divided into high-dose RANKL, low-dose RANKL, OVX, and vehicle control groups. The right proximal tibiae were micro-computed tomography (micro-CT) scanned in vivo every 2 weeks from week 0 to week 12 and every 4 weeks from week 12 to week 20. Bone architectural, mineralization, and mechanical changes were determined. Serum calcium, RANKL, anti-RANKL, and osteoprotegerin were measured at weeks 0, 6, and 20. RESULTS: High-dose RANKL administration resulted in severe deterioration of the trabecular architecture (39% of baseline BV/TV), and modest decreases in tissue mineralization, bone mass, and stiffness. Bone loss occurred more rapidly than in the OVX and low-dose RANKL group, and recovery occurred prior to stopping RANKL injections. Full recovery of trabecular thickness, tissue mineralization, and cortical bone mass, partial recovery of trabecular bone volume (55% of baseline), structural model index, bone mass (69% of baseline), and stiffness (90% of baseline) but no improvement in connectivity density or trabecular number was observed. CONCLUSION: RANKL administration resulted in rapid and dose-dependent bone loss. The recovery of trabecular bone volume and stiffness appeared to be dependent on the number of remaining trabecular elements and their interconnections. Uncontrolled recovery suggests that further investigation into the RANKL-injected rat as a model of bone loss is required.

Implementing a standardized and symptom-oriented flowchart "Kielsflow" for advanced cardiac imaging in a 24/7 interdisciplinary emergency department using spectral CT.

This work focuses on implementing a standardized and symptom-oriented flowchart for advanced cardiac imaging in a 24/7 emergency setting using a dual-layer spectral detector CT system. This flowchart was designed to optimize patient management and standardize imaging workflow. It includes acquisition parameters and contrast agent protocols for the most relevant clinical questions regarding cardiac CT imaging in the interdisciplinary emergency department. The automated reconstruction of symptom-oriented spectral images represents an additional strength here. This implementation is designed to be time-efficient and user-friendly and improves diagnostic quality, independent of the qualification level of clinical and technical personnel.

Time-dependent Viscoelastic Response of Acetabular Bone and Implant Seating during Dynamic Implantation of Press-fit Cups.

Deformation of an acetabular cup implant during cementless implantation is indicative of the radial compressive forces, and such of the initial implant fixation strength. Stress relaxation in the surrounding bone tissue following implantation could reduce the deformation of the cup and thus primary implant fixation. The aim of this study was therefore to determine the early shape change of the implanted cup immediately after implantation with different press-fit levels and whether recording the force during cup impaction can be used to estimate initial cup fixation. Cup implantations into porcine acetabulae (n=10) were performed using a drop tower. The force induced by the drop weight and cup seating after each impact was recorded. Deformation of the implanted cup was determined with strain gauges over a period of 10min. Lever-out torques were measured to assess the initial fixation strength. Stress relaxation in the bone caused a reduction in cup deformation of 13.52±4.06% after 1min and 29.34±5.11% after 10min. The fixation strength increased with a higher force magnitude during impaction (Rs2=0.810, p=0.037). Reduction of the radial compressive forces due to stress relaxation of the surrounding bone should be considered during press-fit cup implantation in order to compensate for the reduced fixation strength over time. In addition, recording the implantation force could help to estimate initial fixation strength.

Individuals with type 2 diabetes mellitus show dimorphic and heterogeneous patterns of loss in femoral bone quality.

Type 2 diabetes mellitus (T2DM), a metabolic disease on the rise, is associated with substantial increase in bone fracture risk. Because individuals with T2DM have normal or high bone mineral density (BMD), osteodensitometric measurements of BMD do not predict fracture risk with T2DM. Here, we aim to identify the underlying mechanism of the diabetes-induced fracture risk using a high-resolution multi-scale analysis of human cortical bone with special emphasis on osseous cellular activity. Specifically, we show increased cortical porosity in a subgroup of T2DM individuals accompanied by changed mineralization patterns and glycoxidative damage to bone protein, caused by non-enzymatic glycation of bone by reducing sugar. Furthermore, the high porosity T2DM subgroup presents with higher regional mineralization heterogeneity and lower mineral maturity, whereas in the T2DM subgroup regional higher mineral-to-matrix ratio was observed. Both T2DM groups show significantly higher carboxymethyl-lysine accumulation. Our results show a dimorphic pattern of cortical bone reorganization in individuals afflicted with T2DM and hence provide new insight into the diabetic bone disease leading to increased fracture risk.

Deformation of acetabular press-fit cups: Influence of design and surgical factors.

BACKGROUND: Deformation of acetabular cups when press-fitted into an undersized cavity is inevitable due to the inhomogeneous stiffness of acetabular bone. Thinner cups or screw holes might increase the risk of high cup deformation. The aim of this study was to examine the influence of cup design and liner assembly on the deformation response during cup implantation. METHODS: Acetabular cups with different designs were implanted into polyurethane foam models simulating the anatomical situation with nominal press-fits of 1mm and without nominal press-fits (line-to-line). Deformations were determined using a tactile coordinate measuring machine. A 3D laser scanner was used to determine the contact conditions at the cup-cavity interface. Polyethylene and ceramic liners were assembled to the implanted cups and the influence of the insertion on the deformation response evaluated. Fixation strength of the cups was determined by push-out testing. FINDINGS: Cup deformation increased with smaller wall thickness (P < 0.037) and screw holes (P < 0.001). Insertion of ceramic liners reduced the deformation (P < 0.001), whereas polyethylene liners adapted to the deformation of the implanted cups (P > 0.999). Thin-walled cups exhibited a higher fixation strength for similar implantation forces (P = 0.011). INTERPRETATION: Thin-walled cups achieved higher fixation strengths and might be more bone-preserving. However, in combination with screw holes and high press-fit levels, wall thickness should be considered carefully to avoid excessive cup deformations leading to potential complications during liner assembly. Line-to-line insertion of thin-walled cups should be accompanied with a rough surface coating to minimize the loss of fixation strength due to the low press-fit fixation.

The influence of cavity preparation and press-fit cup implantation on restoring the hip rotation center.

BACKGROUND: Reaming of the acetabular cavity and cup implantation directly influence the hip rotation center and contact area between implant and bone. Previous studies have reported on an altered rotation center after total hip arthroplasty, but have not studied the influence of reaming and cup implantation separately. Aim of this study was therefore to analyze the individual influence of acetabular reaming and subsequent cup implantation on the rotation center and how this influences the contact conditions at the bone-implant interface. METHODS: Acetabular press-fit cups were implanted into the left and right hips of three full cadavers (n = 6). CT scans were performed to calculate the change in hip rotation center after reaming and prior to liner insertion. 3D models of the cups were used to determine the polar gap, the contact conditions and the effective press-fit. FINDINGS: Reaming the acetabular cavity shifted the rotation center medially (median 5.8 mm, range 4.8-9.1), superiorly (5.3 mm, 3.0-7.0) and posteriorly (2.9 mm, 1.0-5.3). With cup implantation, the rotation center shifted back towards the native position, but no full restoration was observed. The degree of shift increased with the size of polar gap (rs = 0.829, P = .042), which inversely reduced the contact area (rs = 0.886, P = .019). INTERPRETATION: This study reveals that the dominant factor in hip rotation center restoration is the reaming process, while the cup implantation for a given nominal press-fit has only a small influence. Increasing the press-fit would improve the restoration but bares the danger of insufficient bone coverage and periprosthetic fractures due to the high forces needed.

Effect of cavity preparation and bone mineral density on bone-interface densification and bone-implant contact during press-fit implantation of hip stems.

Implant loosening and periprosthetic fracture are two major revision causes for uncemented hip stems. The chosen method of cavity preparation could play a key role for both failure mechanisms. The aim of this study was to determine the dependence of the broach type as well as patient bone mineral density (BMD) on densification and contact conditions at the bone-implant interface. Hip stems were implanted into cadaveric femora using compaction, blunt extraction or sharp extraction broaches with computed tomography scans performed prior to broaching, after broaching and after stem implantation. Proximal periprosthetic bone densification as well as press-fit, contact area and stem seating relative to the last broach were determined. Median bone densification was higher with the compaction and blunt extraction broaches compared to sharp extraction broaches (181% and 177%, respectively, p = 0.002). The bone densification of femora prepared with compaction broaching increased with higher BMD (R2 = 0.183, p = 0.037), while stem seating decreased with higher BMD for all broach types (R2 = 0.259, p = 0.001). Incomplete seated prostheses were associated with smaller press-fit and bone-implant contact area (R2 = 0.249, p = 0.001; R2 = 0.287, p < 0.001). Clinical Significance: The results suggest that compaction broaching maximizes bone densification in patients with higher bone density. However, there appears to be an increased risk of insufficient stem seating in high-density bone that could limit the benefits for primary stability. For lower quality bone, the broach type appears to play a lesser role, but care must be taken to limit extensive stem seating which might increase periprosthetic fracture risk. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:1580-1589, 2019.

Radiation effects on bone architecture in mice and rats resulting from in vivo micro-computed tomography scanning.

Recently established techniques for performing in vivo micro-computed tomography (micro-CT) provide the capability of monitoring bone changes in a living animal at various points in time. However, radiation exposure from repeated micro-CT scans may have an effect on skeletal growth in normal or disease-model animals. The purpose of this study is to test a high resolution (approximately 10 microm) in vivo micro-CT protocol on mice and rats used for bone research to understand the impact of micro-CT radiation exposure on bone architecture. Ovariectomy (OVX) or sham-OVX surgery was performed on groups (n=6-8/group) of 12-week-old C3H/HeJ, C57BL/6J, and BALB/cByJ mice, and one strain of rat (Wistar, retired breeders). The right proximal tibiae were scanned at weekly intervals while the contralateral left limbs were not scanned until the endpoint of the protocol. Trabecular and cortical bone morphology was compared between radiated and non-radiated limbs at the endpoint to quantify the radiation effect. No effects of radiation were observed in OVX or sham rats. Lower trabecular bone volume was observed in the radiated limbs (-8 to -20% relative to non-radiated limb) of all mice groups except sham BALB/cByJ mice and normal control C57BL/6J mice, however, the observed effects were much less than the observed effects of ovariectomy ( approximately 40-50% total bone volume reduction, depending on mouse strain), and no interactions between radiation and OVX treatment were observed (p>0.2). Using an internal non-radiated control within each animal is a potential method to elucidate the effect of radiation exposure for any in vivo protocol. Thus, although in vivo micro-CT is a valuable tool for bone-related research, the impact of radiation in skeletally immature mice should be considered, particularly for strains with low bone volume at the measured site.

Thy-1 Deficiency Augments Bone Loss in Obesity by Affecting Bone Formation and Resorption.

Healthy bone remodeling results from a balanced bone formation and bone resorption realized by bone-forming osteoblasts and bone-resorbing osteoclasts, respectively. Recently, Thy-1 (CD90) was identified as positive regulator of osteoblast differentiation and activation, thus, promoting bone formation while concurrently inhibiting adipogenesis and obesity in mice. Additionally, Thy-1 did not affect bone resorption. An obesity-related co-morbidity that is increasing in prevalence is a disturbed bone formation resulting in an increased fracture risk. The underlying mechanisms of obesity-induced bone alterations are not yet fully elucidated and therefore therapy options for efficient bone-anabolic treatments are limited. Therefore, we investigated the impact of Thy-1 on bone metabolism under obese conditions. Indeed, high fat diet (HFD) induced obese mice lacking Thy-1 (Thy-1-/-) showed increased body fat mass compared to wildtype (WT) mice while bone mass (-38%) and formation (-57%) were decreased as shown by micro-computed tomography (µCT) measurement, histological analysis, and fourier-transform infrared spectroscopy (FTIR). Interestingly, under obese conditions, lack of Thy-1 affected both osteoblast and osteoclast function. Number (-30%) and activity of osteoblasts were decreased in obese Thy-1-/- mice while osteoclast number (+39%) and activity were increased. Facilitated bone marrow fat accumulation (+56%) in obese Thy-1-/- mice compared to obese WT mice was associated with upregulated tumor necrosis factor α (Tnfα, +46%) and colony stimulating factor 1 receptor (Csf1r) expression, strong promoters of osteoclast differentiation. Moreover, lack of Thy-1 was accompanied by a reduction of osteoprotegerin (Tnfrsf11b) expression (-36%), an inhibitor of osteoclast differentiation. Altered Tnfα, Csf1r, and Tnfrsf11b expression might be responsible for elevated osteoclast activity in obese Thy-1-deficient mice. In summary, our findings show that lack of Thy-1 promotes obesity under HFD conditions while concurrently decreasing bone mass and formation. Mechanistic studies revealed that under obese conditions lack of Thy-1 impairs both bone formation and bone resorption.

Tracking the Progression of Osteolytic and Osteosclerotic Lesions in Mice Using Serial In Vivo µCT: Applications to the Assessment of Bisphosphonate Treatment Efficacy.

The metastasis of tumor cells to bone can lead to osteolytic and osteosclerotic lesions, which cause severe, highly-localized bone destruction and abnormal bone apposition, respectively. Accurate quantification of lesion progression is critical to understand underlying mechanisms and assess treatment efficacy; however, standard structural parameters may be insensitive to local changes. We developed methods to quantify osteolytic and osteosclerotic lesions using micro-computed tomography (µCT) within in vivo mouse datasets. Two Balb/c nude datasets were used: (i) bone-homing MDA-MB-231 (osteolytic) cells injected into the left ventricle, treatment with alendronate or vehicle, and weekly µCT (proximal tibia) for 4 weeks, and (ii) MCF7 (osteosclerotic) cells injected into the right tibia and weekly µCT over 12 weeks. After registering images to baseline, osteolytic lesion volume was determined by summing all baseline bone voxels at distances greater than a threshold (150 µm) from the nearest follow-up. Osteosclerotic lesions were determined by measuring the distance from each follow-up surface voxel to the nearest baseline surface and calculating the standard deviation of distance values (SDDT) of the surrounding voxels. Bone mineral density (BMD), bone volume density (BV/TV), and separation (Sp) were determined for comparison. Osteolytic lesions were observed 1 week after tumor cell injection; however, no corresponding BV/TV losses or Sp increases were observed, indicating that standard parameters were unable to detect early metastatic changes. Lesion volume was smaller in the alendronate versus control group (15.0%, p = 0.004 and 18.6%, p = 0.002 of control lesion volume at weeks 3 and 4, respectively). In the osteosclerotic dataset, increased SDDT was observed following injection, providing a potential new measure of osteosclerotic bone apposition. These data show that quantification of local structural change with serial µCT may overcome the limitations of standard mineral and microstructural parameters, and successfully separates metastatic and normal bone turnover. © 2017 American Society for Bone and Mineral Research.

Thy-1 (CD90) promotes bone formation and protects against obesity.

Osteoporosis and obesity result from disturbed osteogenic and adipogenic differentiation and present emerging challenges for our aging society. Because of the regulatory role of Thy-1 in mesenchyme-derived fibroblasts, we investigated the impact of Thy-1 expression on mesenchymal stem cell (MSC) fate between osteogenic and adipogenic differentiation and consequences for bone formation and adipose tissue development in vivo. MSCs from Thy-1-deficient mice have decreased osteoblast differentiation and increased adipogenic differentiation compared to MSCs from wild-type mice. Consistently, Thy-1-deficient mice exhibited decreased bone volume and bone formation rate with elevated cortical porosity, resulting in lower bone strength. In parallel, body weight, subcutaneous/epigonadal fat mass, and bone fat volume were increased. Thy-1 deficiency was accompanied by reduced expression of specific Wnt ligands with simultaneous increase of the Wnt inhibitors sclerostin and dickkopf-1 and an altered responsiveness to Wnt. We demonstrated that disturbed bone remodeling in osteoporosis and dysregulated adipose tissue accumulation in patients with obesity were mirrored by reduced serum Thy-1 concentrations. Our findings provide new insights into the mutual regulation of bone formation and obesity and open new perspectives to monitor and to interfere with the dysregulated balance of adipogenesis and osteogenesis in obesity and osteoporosis.

Automatic segmentation of cortical and trabecular compartments based on a dual threshold technique for in vivo micro-CT bone analysis.

The use of high resolution peripheral quantitative computed tomography (HR-pQCT) and in vivo micro-CT for studies of bone disease and treatment has become increasingly common, and with these methods comes large quantities of data requiring analysis. A simple, robust, and fully-automated segmentation algorithm is presented that efficiently segments bone regions. The dual threshold technique refers to two required threshold inputs that are used to extract the periosteal and endosteal surfaces of the cortex. The proposed method was tested against the gold standard, semi-automated hand contouring, using 45 datasets: mouse, rat, human, and cadaver data from the tibia or radius with nominal isotropic resolutions of 10-82 microm. The performance of the proposed method to segment cortical and trabecular compartments was evaluated qualitatively from visualizations and quantitatively based on morphological measurements. Visual inspection confirmed successful segmentation of all datasets using the new method, with qualitatively better results when applied to the human and cadaver data compared to the gold standard. The dual threshold algorithm was able to extract thin and porous cortices, whereas some clipping and perforations occurred for the gold standard. Morphological parameters measured for segmentation by the proposed method versus the gold standard agreed (95% confidence) for Tb.Th, Tb.Sp, and Tb.N, but not Ct.Th and BV/TV for the human and cadaver datasets. Nonetheless, correlations ranged from 0.95 to 1.00 for all morphological parameters except the cadaver Ct.Th because systematic errors were present. Poor agreement for Ct.Th and BV/TV was due to qualitatively incorrect segmentation by the gold standard when the cortex was thin compared to trabeculae, or operator bias during hand contouring. Since Tb.Th, Tb.Sp, and Tb.N were insensitive to segmentation method, despite operator bias, they are robust parameters for inter-site comparisons. The dual threshold method offers a robust and fully-automated alternative to the gold standard that can efficiently segment bone regions with accurate and repeatable results. The algorithm can be easily implemented since it uses simple image analysis tools. Two input thresholds allow adjustment of the masked output, and are easily determined by trial and error. Using the same input thresholds for similar datasets assures maximal consistency while alleviating time consuming semi-automated contouring.

Establishment of an architecture-specific experimental validation approach for finite element modeling of bone by rapid prototyping and high resolution computed tomography.

A new experimental validation method for assessing the accuracy of large-scale finite element (FE) models of bone micro-structure at the apparent and tissue level was developed. Augmented scaled bone replicas were built using rapid prototype machines based on micro-computed tomography (micro-CT) data. The geometric accuracy of the model was evaluated by comparing experimental tests with the replicas to the FE solution based on the same micro-CT data. A new version of the large-scale FE solver was developed to incorporate orthotropic material properties, hence the experimentally determined properties of the rapid prototype material were input into the FE models. The modified FE solver predicted the experimental apparent level stiffness within less than 1%, and the difference between experimental strain gauge measurements and FE-calculated surface stresses was 7% and 49% on a flat and curved surface region, respectively. While absolute error estimates of surface stresses were limited due to strain gauge errors, the relatively larger difference on the curved surface is indicative of the limitations of a hexahedron FE model for representing such geometries. Although the validation approach is applied here for hexahedron based meshes, the method is flexible for varying bone architectures and will be important for validation of future large-scale FE modeling developments that utilize techniques such as mesh smoothing and tetrahedron elements.

Assessment of Bone Fragility in Patients With Multiple Myeloma Using QCT-Based Finite Element Modeling.

Multiple myeloma (MM) is a malignant plasma cell disease associated with severe bone destruction. Surgical intervention is often required to prevent vertebral body collapse and resulting neurological complications; however, its necessity is determined by measuring lesion size or number, without considering bone biomechanics. Finite element (FE) modeling, which simulates the physiological loading, may improve the prediction of fragility. To test this, we developed a quantitative computed tomography (QCT)-based FE model of the vertebra and applied it to a dataset of MM patients with and without prevalent fracture. FE models were generated from vertebral QCT scans of the T12 (T11 if T12 was fractured) of 104 MM patients, 45 with fracture and 59 without, using a low-dose scan protocol (1.5 mm slice thickness, 4.0 to 6.5 mSv effective dose). A calibration phantom enabled the conversion of the CT Hounsfield units to FE material properties. Compressive loading of the vertebral body was simulated and the stiffness, yield load, and work to yield determined. To compare the parameters between fracture and nonfracture groups, t tests were used, and standardized odds ratios (sOR, normalized to standard deviation) and 95% confidence intervals were calculated. FE parameters were compared to mineral and structural parameters using linear regression. Patients with fracture showed lower vertebral stiffness (-15.2%; p = 0.010; sOR = 1.73; 95% CI, 1.11 to 2.70), yield force (-21.5%; p = 0.002; sOR = 2.09; 95% CI, 1.27 to 3.43), and work to yield (-27.4%; p = 0.001; sOR = 2.28; 95% CI, 1.33 to 3.92) compared to nonfracture patients. All parameters correlated significantly with vBMD (stiffness: R2 = 0.57, yield force: R2 = 0.59, work to yield: R2 = 0.50, p < 0.001), BV/TV (stiffness: R2 = 0.56, yield force: R2 = 0.58, work to yield: R2 = 0.49, p < 0.001), and Tb.Sp (stiffness: R2 = 0.51, yield force: R2 = 0.53, work to yield: R2 = 0.45, p < 0.001). FE modeling identified MM patients with compromised mechanical integrity of the vertebra. Higher sOR values were obtained for the biomechanical compared to structural or mineral measures, suggesting that FE modeling improves fragility assessment in these patients. © 2016 American Society for Bone and Mineral Research.