Microcomputed Tomography Is a Precise Method That Allows for Topographical Characterization of Lymph Nodes and Lymphatic Vessels.

Background: Surgical excision and/or radiation targeting of regional lymph nodes are an essential component in the clinical management of cancer. Importantly, a more accurate understanding of lymphatic anatomy could enable refinement of present treatment strategies. Given the spatial resolution limitations of contemporary imaging methods, our group sought to utilize noncontrast-enhanced microcomputed tomography (µCT) imaging to clarify regional lymphatic anatomy. Methods and Results: This study was conducted with embalmed en bloc lymphatic tissue packets from six donors (three females and three males: medianage of death = 78 years). All specimens were investigated with noncontrast-enhanced µCT imaging using a conebeam-CT imaging system. Adipose and lymphatic tissues were segmented by radiodensity based on sampling regions of interest. To confirm the observations from µCT, lymph nodes from each packet were exposed to hematoxylin and eosin staining and anti-D240 immunostaining. Following µCT imaging, mean peak radiodensities of -203.14 ± 19.35 Hounsfield units (HU) and 37.25 ± 31.95 HU were revealed for adipose and lymphatic tissues, respectively (p < 0.01). By analyzing histograms of the radiodensity distributions, we determined a threshold of -82.42 HU to differentiate adipose and lymphatic tissue, to generate three-dimensional renderings, and to calculate quantitative metrics. On average, adipose tissue comprised 9.62 ± 3.60 cm3 (73.6%) of the total packet volume, whereas lymphatic tissue comprised 3.47 ± 2.71 cm3 (26.4%). Moreover, each en bloc packet contained four small lymph nodes (1-5 mm) and three to four large lymph nodes (>5 mm). Histology corroborated the observations from µCT. Conclusions: Altogether, a precise understanding of regional lymphatic anatomy elucidated by the present imaging modality may help refine clinical cancer treatment strategies.

Ectopic spinal calcification associated with diffuse idiopathic skeletal hyperostosis (DISH): A quantitative micro-ct analysis.

Diffuse idiopathic skeletal hyperostosis (DISH) is a non-inflammatory spondyloarthropathy identified radiographically by calcification of the ligaments and/or entheses along the anterolateral aspect of the vertebral column. The etiology and pathogenesis of calcifications are unknown, and the diagnosis of DISH is currently based on radiographic criteria associated with advanced disease. To characterize the features of calcifications associated with DISH, we used micro-computed tomographic imaging to evaluate a cohort of 19 human cadaveric vertebral columns. Fifty-three percent of the cohort (n = 10; 3 females, 7 males, mean age of death = 81 years, range 67-94) met the radiographic criteria for DISH, with calcification of four or more contiguous vertebral segments. In almost all cases, the lower thoracic regions (T8-12) were affected by calcifications, consisting primarily of large, horizontal outgrowths of bony material. In contrast, calcifications localized to the upper thoracic regions demonstrated variability in their presentation and were categorized as either "continuous vertical bands" or "discontinuous-patchy" lesions. In addition to the variable morphology of the calcifications, our analysis demonstrated remarkable heterogeneity in the densities of calcifications, ranging from internal components below the density of cortical bone to regions of hyper-dense material that exceeded cortical bone. These findings establish that the current radiographic criteria for DISH capture heterogeneous presentations of ectopic spine calcification that can be differentiated based on morphology and density. These findings may indicate a naturally heterogenous disease, potential stage(s) in the natural progression of DISH, or distinct pathologies of ectopic calcifications. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res.

In-vitro comparison of different palatal sites for orthodontic miniscrew insertion: Effect of bone quality and quantity on primary stability.

INTRODUCTION: This experiment was undertaken to assess the primary stability of orthodontic miniscrews inserted at different sites in human cadaveric palatal bone for temporary skeletal anchorage, and to determine the effect of bone quality and quantity on their primary stability using microcomputed tomography imaging. METHODS: A total of 10 cadaveric maxillary hard palates were used for insertion of 130 orthodontic miniscrews (VectorTAS; Ormco, Orange, Calif; length, 6 mm; diameter, 1.4 mm). Upon insertion, maximal insertion torque (IT) was recorded. Imaging (microcomputed tomography) was performed before and after insertion for assessment of bone quality and quantity parameters (bone mineral density [BMD], bone thickness [BT], and length of screw engagement [LSE]). Differences in each parameter were assessed at the various insertion sites. Correlations between IT and measurements of BMD, BT, and LSE were evaluated. RESULTS: Significant differences (P < 0.001) were found among insertion sites for IT, BT, and LSE, but not for BMD (P = 0.004). Correlations were found between IT and BMD (rs = 0.42; P < 0.001), IT and BT (rs = 0.58; P < 0.001), and IT and LSE (rs = 0.58; P < 0.001). Most perforations of miniscrews into the nasal cavity occurred posterior to the permanent second premolars. CONCLUSIONS: The primary stability of orthodontic miniscrews in the palate is affected by bone quality and quantity, with higher primary stability obtained anterior to the second premolars and parasagittally at the level of the permanent first molars.

Quantitative in vivo micro-computed tomography for assessment of age-dependent changes in murine whole-body composition.

Micro-computed tomography (micro-CT) is used routinely to quantify skeletal tissue mass in small animal models. Our goal was to evaluate repeated in vivo micro-CT imaging for monitoring whole-body composition in studies of growth and aging in mice. Male mice from 2 to 52 weeks of age were anesthetized and imaged using an eXplore Locus Ultra and/or eXplore speCZT scanner. Images were reconstructed into 3D volumes, signal-intensity thresholds were used to classify each voxel as adipose, lean or skeletal tissue, and tissue masses were calculated from known density values. Images revealed specific changes in tissue distribution with growth and aging. Quantification showed biphasic increases in total CT-derived body mass, lean and skeletal tissue masses, consisting of rapid increases to 8 weeks of age, followed by slow linear increases to 52 weeks. In contrast, bone mineral density increased rapidly to a stable plateau at ~ 14 weeks of age. On the other hand, adipose tissue mass increased continuously with age. A micro-CT-derived total mass was calculated for each mouse and compared with gravimetrically measured mass, which differed on average by < 3%. Parameters were highly reproducible for mice of the same age, but variability increased slightly with age. There was also good agreement in parameters for the same group of mice scanned on the eXplore Locus Ultra and eXplore speCZT systems. This study provides reference values for normative comparisons; as well, it demonstrates the usefulness of in vivo single-energy micro-CT scans to quantify whole-body composition in high-throughput studies of growth and aging in mice.

Quantification of mouse in vivo whole-body vibration amplitude from motion-blur using x-ray imaging.

Musculoskeletal effects of whole-body vibration on animals and humans have become an intensely studied topic recently, due to the potential of applying this method as a non-pharmacological therapy for strengthening bones. It is relatively easy to quantify the transmission of whole-body mechanical vibration through the human skeletal system using accelerometers. However, this is not the case for small-animal pre-clinical studies because currently available accelerometers have a large mass, relative to the mass of the animals, which causes the accelerometers themselves to affect the way vibration is transmitted. Additionally, live animals do not typically remain motionless for long periods, unless they are anesthetized, and they are required to maintain a static standing posture during these studies. These challenges provide the motivation for the development of a method to quantify vibrational transmission in small animals. We present a novel imaging technique to quantify whole-body vibration transmission in small animals using 280 µm diameter tungsten carbide beads implanted into the hind limbs of mice. Employing time-exposure digital x-ray imaging, vibrational amplitude is quantified based on the blurring of the implanted beads caused by the vibrational motion. Our in vivo results have shown this technique is capable of measuring vibration amplitudes as small as 0.1 mm, with precision as small as ±10 µm, allowing us to distinguish differences in the transmitted vibration at different locations on the hindlimbs of mice.

Loss of P2X7 nucleotide receptor function leads to abnormal fat distribution in mice.

The P2X7 receptor is an ATP-gated cation channel expressed by a number of cell types. We have shown previously that disruption of P2X7 receptor function results in downregulation of osteogenic markers and upregulation of adipogenic markers in calvarial cell cultures. In the present study, we assessed whether loss of P2X7 receptor function results in changes to adipocyte distribution and lipid accumulation in vivo. Male P2X7 loss-of-function (KO) mice exhibited significantly greater body weight and epididymal fat pad mass than wild-type (WT) mice at 9 months of age. Fat pad adipocytes did not differ in size, consistent with adipocyte hyperplasia rather than hypertrophy. Histological examination revealed ectopic lipid accumulation in the form of adipocytes and/or lipid droplets in several non-adipose tissues of older male KO mice (9-12 months of age). Ectopic lipid was observed in kidney, extraorbital lacrimal gland and pancreas, but not in liver, heart or skeletal muscle. Specifically, lacrimal gland and pancreas from 12-month-old male KO mice had greater numbers of adipocytes in perivascular, periductal and acinar regions. As well, lipid droplets accumulated in the renal tubular epithelium and lacrimal acinar cells. Blood plasma analyses revealed diminished total cholesterol levels in 9- and 12-month-old male KO mice compared with WT controls. Interestingly, no differences were observed in female mice. Moreover, there were no significant differences in food consumption between male KO and WT mice. Taken together, these data establish novel in vivo roles for the P2X7 receptor in regulating adipogenesis and lipid metabolism in an age- and sex-dependent manner.

Least-error projection sorting to optimize retrospectively gated cardiac micro-CT of free-breathing mice.

PURPOSE: To develop and characterize a technique for optimizing image quality by eliminating streaking artifacts in retrospectively gated microcomputed tomography (micro-CT) images of mice caused by insufficient and irregular angular sampling. METHODS: A least-error sorting technique was developed to minimize streak artifacts in retrospectively gated cardiac micro-CT images. To ensure complete filling of projection space, for each angular position, the projection acquired closest to the desired cardiac phase is used to reconstruct a volumetric image. An acrylic slanted-edge phantom undergoing cyclic motion was used to characterize the system's resolution. The phantom was scanned using a volumetric micro-CT scanner equipped with a flat-panel detector mounted on a slip-ring gantry. Projection images of the moving phantom were collected over a period of 60 s using a variety of acquisition protocols with the rotation period of the gantry ranging from 1 to 5 s. The modulation transfer function (MTF) of the reconstructed images was measured for many combinations of acquisition and reconstruction parameters. The use of the least-error technique was also demonstrated in vivo. RESULTS: The motion blurring introduced into the images at physiologically significant velocities of 6 cm∕s agreed well with predicted values; limiting resolution (frequency at 10% MTF) degraded from 2.5 to 1.0 mm(-1) for a velocity of 6 cm∕s and 5 s∕rotation gantry speed. Faster gantry rotation speeds led to improved temporal resolution but the scanner's data storage and transfer rates and field of view limitations made scanning at gantry speeds faster than 2 s∕rotation impractical. CONCLUSIONS: The least-error technique effectively eliminates streaking artifact caused by missing views and allows for optimization of image quality in retrospectively gated micro-CT.

Subchondral cysts create increased intra-osseous stress in early knee OA: A finite element analysis using simulated lesions.

AIM OF STUDY: To investigate the role of intra-osseous lesions in advancing the pathogenesis of Osteoarthritis (OA) of the knee, using Finite Element Modeling (FEM) in conjunction with high-resolution imaging techniques. METHODS: Twenty early stage OA patients (≤ Grade 2 radiographic score) were scanned with a prototype, cone-beam CT system. Scans encompassed the mid-shaft of the femur to the diaphysis of the proximal tibia. Individual bones were segmented to create 3D geometric models that were transferred to FE software for loading experiments. Patient-specific, inhomogeneous material properties were derived from the CT images and mapped directly to the FE models. Duplicate models were also created, with a 3D sphere (range 3-12 mm) introduced into a weight-bearing region of the joint, mimicking the size, location, and composition of a subchondral bone cyst (SBC). A spherical shell extending 1mm radially around the SBC served as the sample volume for measurements of von Mises equivalent stress. Both models were vertically loaded with 750 N, or approximately 1 body weight during a single-leg stance. RESULTS: All FE models exhibited a physiologically realistic weight-bearing distribution of stress, which initiated at the joint surface and extended to the cortical bone. Models that contained the SBC experienced a nearly two-fold increase in stress (0.934 ± 0.073 and 1.69 ± 0.159 MPa, for the non-SBC and SBC models, respectively) within the bone adjacent to the SBC. In addition, there was a positive correlation found between the diameter of the SBC and the resultant intra-osseous stress under load (p = 0.004). CONCLUSIONS: Our results provide insights into the mechanism by which SBC may accelerate OA, leading to greater pain and disability. Based on these findings, we feel that patient-derived FE models of the OA knee - utilizing in vivo imaging data - present a tremendous potential for monitoring joint mechanics under physiological loads.

Error analysis of marker-based object localization using a single-plane XRII.

The role of imaging and image guidance is increasing in surgery and therapy, including treatment planning and follow-up. Fluoroscopy is used for two-dimensional (2D) guidance or localization; however, many procedures would benefit from three-dimensional (3D) guidance or localization. Three-dimensional computed tomography (CT) using a C-arm mounted x-ray image intensifier (XRII) can provide high-quality 3D images; however, patient dose and the required acquisition time restrict the number of 3D images that can be obtained. C-arm based 3D CT is therefore limited in applications for x-ray based image guidance or dynamic evaluations. 2D-3D model-based registration, using a single-plane 2D digital radiographic system, does allow for rapid 3D localization. It is our goal to investigate-over a clinically practical range-the impact of x-ray exposure on the resulting range of 3D localization precision. In this paper it is assumed that the tracked instrument incorporates a rigidly attached 3D object with a known configuration of markers. A 2D image is obtained by a digital fluoroscopic x-ray system and corrected for XRII distortions (+/- 0.035 mm) and mechanical C-arm shift (+/- 0.080 mm). A least-square projection-Procrustes analysis is then used to calculate the 3D position using the measured 2D marker locations. The effect of x-ray exposure on the precision of 2D marker localization and on 3D object localization was investigated using numerical simulations and x-ray experiments. The results show a nearly linear relationship between 2D marker localization precision and the 3D localization precision. However, a significant amplification of error, nonuniformly distributed among the three major axes, occurs, and that is demonstrated. To obtain a 3D localization error of less than +/- 1.0 mm for an object with 20 mm marker spacing, the 2D localization precision must be better than +/- 0.07 mm. This requirement was met for all investigated nominal x-ray exposures at 28 cm FOV, and for all but the lowest two at 40 cm FOV. However, even for those two nominal exposures, the expected 3D localization error is less than +/- 1.2 mm. The tracking precision was +/- 0.65 mm for the out-of-plane translations, +/- 0.05 mm for in-plane translations, and +/- 0.05 degrees for the rotations. The root mean square (RMS) difference between the true and projection-Procrustes calculated location was 1.07 mm. It is believed these results show the potential of this technique for dynamic evaluations or real-time image guidance using a single x-ray source and XRII detector.

Four dimensional intravenous cone-beam computed tomographic subtraction angiography. In vitro study of feasibility.

OBJECTIVE: We demonstrate the feasibility of 4D intravenous computed tomographic (CT) subtraction cerebral angiography using in vitro, anthropomorphic techniques. MATERIALS AND METHODS: High-resolution 3D cone-beam CT datasets (0.45 mm isotropic voxel size, 120 kVp, 90 mA) of a cadaver-derived cerebrovascular phantom, containing a saccular aneurysm, were acquired at a rate of 1 Hz for 20 seconds. A computer-controlled pump provided physiologically realistic blood-flow waveforms using a water-glycerol blood-mimicking fluid (10 mL/s mean flow). Contrast agent injected at 0.94 mL/s for 5 seconds provided a clinically realistic intravenous vascular enhancement of approximately 300 Hounsfield units. The first 4 to 5 volumes (precontrast) provided a mask dataset for volumetric subtraction. Vascular enhancement was measured in the dynamic, time-resolved, subtracted 3D angiograms. Contrast-to-noise ratio was measured in 3D source data and maximum intensity projections (MIPs). Dose measurements were made using an ionization chamber. RESULTS: MIP images of the time-resolved volumetric data were of diagnostic quality, clearly showing the aneurysm dome and neck, and cerebral vessels. Dynamic flow information (contrast wash-in/wash-out) was observed, including differential opacification and draining of the anterior and posterior vasculature and the aneurysm. Contrast-to-noise ratio was measured to be in the range of 3 to 4.5 in averaged volumes, and 10.5 to 17 in the corresponding MIPs, at an effective patient dose of 2.8 mSv, with 4 cm of axial coverage. CONCLUSIONS: We have demonstrated the feasibility of 4D volumetric, intravenous CT subtraction angiography, in vitro, providing time-resolved, diagnostic quality 3D datasets. We were able to show time-resolved blood-flow information and high-resolution local and global anatomic renderings, from a single 20-second scan, at acceptable x-ray dose.

A quality assurance phantom for the performance evaluation of volumetric micro-CT systems.

Small-animal imaging has recently become an area of increased interest because more human diseases can be modeled in transgenic and knockout rodents. As a result, micro-computed tomography (micro-CT) systems are becoming more common in research laboratories, due to their ability to achieve spatial resolution as high as 10 microm, giving highly detailed anatomical information. Most recently, a volumetric cone-beam micro-CT system using a flat-panel detector (eXplore Ultra, GE Healthcare, London, ON) has been developed that combines the high resolution of micro-CT and the fast scanning speed of clinical CT, so that dynamic perfusion imaging can be performed in mice and rats, providing functional physiological information in addition to anatomical information. This and other commercially available micro-CT systems all promise to deliver precise and accurate high-resolution measurements in small animals. However, no comprehensive quality assurance phantom has been developed to evaluate the performance of these micro-CT systems on a routine basis. We have designed and fabricated a single comprehensive device for the purpose of performance evaluation of micro-CT systems. This quality assurance phantom was applied to assess multiple image-quality parameters of a current flat-panel cone-beam micro-CT system accurately and quantitatively, in terms of spatial resolution, geometric accuracy, CT number accuracy, linearity, noise and image uniformity. Our investigations show that 3D images can be obtained with a limiting spatial resolution of 2.5 mm(-1) and noise of +/-35 HU, using an acquisition interval of 8 s at an entrance dose of 6.4 cGy.

Correction of XRII geometric distortion using a liquid-filled grid and image subtraction.

X-ray image intensifier (XRII) geometric distortion reduces the accuracy of image-guided procedures and quantitative image reconstructions. Due to the dependence of this error on the earth's magnetic field, the required correction is angle dependent, and calibration data should ideally be acquired simultaneously with clinical image data, at a specific orientation. We describe a technique to correct XRII geometric image distortion at any angular position during a stereotactic procedure. This approach uses a machined plastic grid, which contains channels that can be filled with iodinated contrast agent and subsequently flushed with water, providing contrast and mask images, respectively, of a geometric calibration grid. The standard image subtraction capabilities of conventional digital subtraction angiography devices can then be used to create a subtraction image of the iodine-filled channels, without any confounding anatomical structure. Grid-line intersection points are used to determine the control points that are required for a global polynomial correction algorithm, creating a correction map that is specific to the current angular position and XRII field of view (FOV). Tests with a clinical C-arm based XRII show that control points can be obtained with a precision of +/-0.053 mm, resulting in geometric correction accuracy of +/-0.152 mm, at a nominal FOV of 40 cm. While the precision and accuracy are both poorer than that achieved with a high-contrast steel-bead grid, the fact that the liquid grid can remain rigidly attached to the XRII during an entire procedure results in the establishment of an absolute detector coordinate system (referenced to the liquid-filled correction grid). The design of the liquid-filled channels allows the required control points to be introduced into the image or removed in about 30 s, avoiding the appearance of obscuring or confounding markers during clinical image acquisition, with a concurrent increase in patient dose of about 8% in the current design. Applications for this technique include stereotactic surgery, radiosurgery, x-ray stereogrammetry, and other image-guided procedures.