High tibial osteotomy to neutral alignment improves medial knee articular cartilage composition.

PURPOSE: The purpose of this study was to: (1) test the hypothesis that HTO improves articular cartilage composition in the medial compartment without adversely affecting the lateral compartment and patella, and; (2) explore associations between knee alignment and cartilage composition after surgery. METHODS: 3T MRI and standing radiographs were obtained from 34 patients before and 1-year after HTO. Articular cartilage was segmented from T2 maps. Mechanical axis angle (MAA), posterior tibial slope, and patellar height were measured from radiographs. Changes in T2 and radiographic measures were assessed using paired t tests, and associations were assessed using Pearson correlation coefficients. RESULTS: The mean (SD) MAA before and after HTO was - 6.5° (2.4) and 0.6° (3.0), respectively. There was statistically significant shortening [mean (95%CI)] of T2 in the medial femur [- 2.8 ms (- 4.2; - 1.3), p < 0.001] and medial tibia [- 2.2 ms (- 3.3; - 1.0), p < 0.001], without changes in the lateral femur [- 0.5 ms (- 1.6; 0.6), p = 0.3], lateral tibia [0.2 ms (- 0.8; 1.1), p = NS], or patella [0.5 ms (- 1.0; 2.1), p = NS). Associations between radiographic measures and T2 were low. 23% of the increase in lateral femur T2 was explained by postoperative posterior tibial slope (r = 0.48). CONCLUSION: Performing medial opening wedge HTO without overcorrection improves articular cartilage composition in the medial compartment of the knee without compromising the lateral compartment or the patella. Although further research is required, these results suggest HTO is a disease structure-modifying treatment for knee OA.

Extending the dynamic range of biomedical micro-computed tomography for application to geomaterials.

BACKGROUND: X-ray computed tomography (CT) can non-destructively examine objects by producing three-dimensional images of their internal structure. Although the availability of biomedical micro-CT offers the increased access to scanners, CT images of dense objects are susceptible to artifacts particularly due to beam hardening. OBJECTIVE: This study proposes and evaluates a simple semi-empirical correction method for beam hardening and scatter that can be applied to biomedical scanners. METHODS: Novel calibration phantoms of varying diameters were designed and built from aluminum and poly[methyl-methacrylate]. They were imaged using two biomedical micro-CT scanners. Absorbance measurements made through different phantom sections were fit to polynomial and inversely exponential functions and used to determine linearization parameters. Corrections based on the linearization equations were applied to the projection data before reconstruction. RESULTS: Correction for beam hardening was achieved when applying both scanners with the correction methods to all test objects. Among them, applying polynomial correction method based on the aluminum phantom provided the best improvement. Correction of sample data demonstrated a high agreement of percent-volume composition of dense metallic inclusions between using the Bassikounou meteorite from the micro-CT images (13.7%) and previously published results using the petrographic thin sections (14.6% 8% metal and 6.6% troilite). CONCLUSIONS: Semi-empirical linearization of X-ray projection data with custom calibration phantoms allows accurate measurements to be obtained on the radiodense samples after applying the proposed correction method on biomedical micro-CT images.

Receptor for hyaluronan mediated motility (RHAMM/HMMR) is a novel target for promoting subcutaneous adipogenesis.

Hyaluronan, CD44 and the Receptor for Hyaluronan-Mediated Motility (RHAMM, gene name HMMR) regulate stem cell differentiation including mesenchymal progenitor differentiation. Here, we show that CD44 expression is required for subcutaneous adipogenesis, whereas RHAMM expression suppresses this process. We designed RHAMM function blocking peptides to promote subcutaneous adipogenesis as a clinical and tissue engineering tool. Adipogenic RHAMM peptides were identified by screening for their ability to promote adipogenesis in culture assays using rat bone marrow mesenchymal stem cells, mouse pre-adipocyte cell lines and primary human subcutaneous pre-adipocytes. Oil red O uptake into fat droplets and adiponectin production were used as biomarkers of adipogenesis. Positive peptides were formulated in either collagen I or hyaluronan (Orthovisc) gels then assessed for their adipogenic potential in vivo following injection into dorsal rat skin and mammary fat pads. Fat content was quantified and characterized using micro CT imaging, morphometry, histology, RT-PCR and ELISA analyses of adipogenic gene expression. Injection of screened peptides increased dorsal back subcutaneous fat pad area (208.3 ± 10.4 mm2versus control 84.11 ± 4.2 mm2; p < 0.05) and mammary fat pad size (45 ± 11 mg above control background, p = 0.002) in female rats. This effect lasted >5 weeks as detected by micro CT imaging and perilipin 1 mRNA expression. RHAMM expression suppresses while blocking peptides promote expression of PPARγ, C/EBP and their target genes. Blocking RHAMM function by peptide injection or topical application is a novel and minimally invasive method for potentially promoting subcutaneous adipogenesis in lipodystrophic diseases and a complementary tool to subcutaneous fat augmentation techniques.

C57BL/6 mice are resistant to joint degeneration induced by whole-body vibration.

OBJECTIVE: Whole-body vibration (WBV) platforms are commercially available devices that are used clinically to treat numerous musculoskeletal conditions based on their reported ability to increase bone mineral density and muscle strength. Despite widespread use, there is an alarming lack of understanding of the direct effects of WBV on joint health. Previous work by our lab demonstrated that repeated exposure to WBV using protocols that model those used clinically, induces intervertebral disc (IVD) degeneration and osteoarthritis-like damage in the knee of skeletally mature, male mice of a single outbred strain (CD-1). The present study examined whether exposure to WBV induces similar deleterious effects in a genetically different strain of mouse (C57BL/6). DESIGN: Male 10-week-old C57BL/6 mice were exposed to vertical sinusoidal WBV for 30 min/day, 5 days/week, for 4 or 8 weeks using previously reported protocols (45 Hz, 0.3 g peak acceleration). Following WBV, joint tissues were examined using histological analysis and gene expression was quantified using real-time PCR (qPCR). RESULTS: Our analyses show a lack of WBV-induced degeneration in either the knee or IVDs of C57BL/6 mice exposed to WBV for 4 or 8 weeks, in direct contrast to the WBV-induced damage previously reported by our lab in CD-1 mice. CONCLUSIONS: Together with previous studies from our group, the present study demonstrates that the effects of WBV on joint tissues vary in a strain-specific manner. These findings highlight the need to examine genetic or physiological differences that may underlie susceptibility to the deleterious effects of WBV on joint tissues.

Whole-body vibration of mice induces articular cartilage degeneration with minimal changes in subchondral bone.

OBJECTIVE: Low-amplitude, high-frequency whole-body vibration (WBV) has been adopted for the treatment of musculoskeletal diseases including osteoarthritis (OA); however, there is limited knowledge of the direct effects of vibration on joint tissues. Our recent studies revealed striking damage to the knee joint following exposure of mice to WBV. The current study examined the effects of WBV on specific compartments of the murine tibiofemoral joint over 8 weeks, including microarchitecture of the tibia, to understand the mechanisms associated with WBV-induced joint damage. DESIGN: Ten-week-old male CD-1 mice were exposed to WBV (45 Hz, 0.3 g peak acceleration; 30 min/day, 5 days/week) for 4 weeks, 8 weeks, or 4 weeks WBV followed by 4 weeks recovery. The knee joint was evaluated histologically for tissue damage. Architecture of the subchondral bone plate, subchondral trabecular bone, primary and secondary spongiosa of the tibia was assessed using micro-CT. RESULTS: Meniscal tears and focal articular cartilage damage were induced by WBV; the extent of damage increased between 4 and 8-week exposures to WBV. WBV did not alter the subchondral bone plate, or trabecular bone of the tibial spongiosa; however, a transient increase was detected in the subchondral trabecular bone volume and density. CONCLUSIONS: The lack of WBV-induced changes in the underlying subchondral bone suggests that damage to the articular cartilage may be secondary to the meniscal injury we detected. Our findings underscore the need for further studies to assess the safety of WBV in the human population to avoid long-term joint damage.

Iodine potassium iodide improves the contrast-to-noise ratio of micro-computed tomography images of the human middle ear.

High-resolution imaging of middle-ear geometry is necessary for finite-element modeling. Although micro-computed tomography (microCT) is widely used because of its ability to image bony structures of the middle ear, it is difficult to visualize soft tissues - including the tympanic membrane and the suspensory ligaments/tendons - because of lack of contrast. The objective of this research is to quantitatively evaluate the efficacy of iodine potassium iodide (IKI) solution as a contrast agent. Six human temporal bones were used in this experiment, which were obtained in right-left pairs, from three cadaveric heads. All bones were fixed using formaldehyde. Three bones (one from each pair) were stained in IKI solution for 2 days, whereas the other three were not stained. Samples were scanned using a microCT system at a resolution of 20 µm. Eight soft tissues in the middle ear were segmented: anterior mallear ligament, incudomallear joint, lateral mallear ligament, posterior incudal ligament, stapedial annular ligament, stapedius muscle, tympanic membrane and tensor tympani muscle. Contrast-to-noise ratios (CNRs) of each soft tissue were calculated for each temporal bone. Combined CNRs of the soft tissues in unstained samples were 6.1 ± 3.0, whereas they were 8.1 ± 2.7 in stained samples. Results from Welch's t-test indicate significant difference between the two groups at a 95% confidence interval. Results for paired t-tests for each of the individual soft tissues also indicated significant improvement of contrast in all tissues after staining. Relatively large soft tissues in the middle ear such as the tympanic membrane and the tensor tympani muscle were impacted by staining more than smaller tissues such as the stapedial annular ligament. The increase in contrast with IKI solution confirms its potential application in automatic segmentation of the middle-ear soft tissues.

Mechanical Forces Exacerbate Periodontal Defects in Bsp-null Mice.

Bone sialoprotein (BSP) is an acidic phosphoprotein with collagen-binding, cell attachment, and hydroxyapatite-nucleating properties. BSP expression in mineralized tissues is upregulated at onset of mineralization. Bsp-null (Bsp(-/-)) mice exhibit reductions in bone mineral density, bone turnover, osteoclast activation, and impaired bone healing. Furthermore, Bsp(-/-) mice have marked periodontal tissue breakdown, with a lack of acellular cementum leading to periodontal ligament detachment, extensive alveolar bone and tooth root resorption, and incisor malocclusion. We hypothesized that altered mechanical stress from mastication contributes to periodontal destruction observed in Bsp(-/-) mice. This hypothesis was tested by comparing Bsp(-/-) and wild-type mice fed with standard hard pellet diet or soft powder diet. Dentoalveolar tissues were analyzed using histology and micro-computed tomography. By 8 wk of age, Bsp(-/-) mice exhibited molar and incisor malocclusion regardless of diet. Bsp(-/-) mice with hard pellet diet exhibited high incidence (30%) of severe incisor malocclusion, 10% lower body weight, 3% reduced femur length, and 30% elevated serum alkaline phosphatase activity compared to wild type. Soft powder diet reduced severe incisor malocclusion incidence to 3% in Bsp(-/-) mice, supporting the hypothesis that occlusal loading contributed to the malocclusion phenotype. Furthermore, Bsp(-/-) mice in the soft powder diet group featured normal body weight, long bone length, and serum alkaline phosphatase activity, suggesting that tooth dysfunction and malnutrition contribute to growth and skeletal defects reported in Bsp(-/-) mice. Bsp(-/-) incisors also erupt at a slower rate, which likely leads to the observed thickened dentin and enhanced mineralization of dentin and enamel toward the apical end. We propose that the decrease in eruption rate is due to a lack of acellular cementum and associated defective periodontal attachment. These data demonstrate the importance of BSP in maintaining proper periodontal function and alveolar bone remodeling and point to dental dysfunction as causative factor of skeletal defects observed in Bsp(-/-) mice.

Deficiency in acellular cementum and periodontal attachment in bsp null mice.

Bone sialoprotein (BSP) is an extracellular matrix protein found in mineralized tissues of the skeleton and dentition. BSP is multifunctional, affecting cell attachment and signaling through an RGD integrin-binding region, and acting as a positive regulator for mineral precipitation by nucleating hydroxyapatite crystals. BSP is present in cementum, the hard tissue covering the tooth root that anchors periodontal ligament (PDL) attachment. To test our hypothesis that BSP plays an important role in cementogenesis, we analyzed tooth development in a Bsp null ((-/-)) mouse model. Developmental analysis by histology, histochemistry, and SEM revealed a significant reduction in acellular cementum formation on Bsp (-/-) mouse molar and incisor roots, and the cementum deposited appeared hypomineralized. Structural defects in cementum-PDL interfaces in Bsp (-/-) mice caused PDL detachment, likely contributing to the high incidence of incisor malocclusion. Loss of BSP caused progressively disorganized PDL and significantly increased epithelial down-growth with aging. Bsp (-/-) mice displayed extensive root and alveolar bone resorption, mediated by increased RANKL and the presence of osteoclasts. Results collected here suggest that BSP plays a non-redundant role in acellular cementum formation, likely involved in initiating mineralization on the root surface. Through its importance to cementum integrity, BSP is essential for periodontal function.

Damage of an Oxinium femoral head and polyethylene liner following 'routine' total hip replacement.

We present a case of early retrieval of an Oxinium femoral head and corresponding polyethylene liner where there was significant surface damage to the head and polyethylene. The implants were retrieved at the time of revision surgery to correct leg-length discrepancy just 48 hours after the primary hip replacement. Appropriate analysis of the retrieved femoral head demonstrated loss of the Oxinium layer with exposure of the underlying substrate and transfer of titanium from the acetabular shell at the time of a reduction of the index total hip replacement. In addition, the level of damage to the polyethylene was extensive despite only 48 hours in situ. The purpose of this report is to highlight the care that is required at the time of reduction, especially with these hard femoral counter-faces such as Oxinium. To our knowledge, the damage occurring at the time of reduction has not been previously reported following the retrieval of an otherwise well-functioning hip replacement.

Design and construction of a multipath vessel phantom for interventional training.

This short communication reports on the design and construction of a catheter manipulation skill enhancement phantom for use by residents and fellows outside the clinical environment. The phantom contains a variety of path trajectories and vessel diameter transitions, allowing trainees to manipulate catheters through vessel paths of varying difficulty. The multipath phantom, which is easy to construct and provides easily visualised paths, provides a simple, cost-effective training platform to facilitate and accelerate interventional training.

Development of a customized density-modulus relationship for use in subject-specific finite element models of the ulna.

Assigning an appropriate density-modulus relationship is an important factor when applying inhomogeneous material properties to finite element models of bone. The purpose of this study was to develop a customized density-modulus equation for the distal ulna, using beam theory combined with experimental results. Five custom equations of the form E= ap(b) were used to apply material properties to models of eight ulnae. All equations passed through a point (1.85, Ec), where p = 1.85 g/cm3 represents the average density of cortical bone. For custom equations (1) to (3), Ec was predicted using beam theory, and the value of b was varied within the range reported in the literature. Custom equations (4) and (5) used other values of Ec from the literature, while keeping b constant. Results obtained from the custom equations were compared with those from other equations in the literature, and with experimental results. The beam theory analysis predicted Ec = 21 +/- 1.6 GPa, and the three custom equations using this value tended to have the lowest errors. The power of the equations did not affect the results as much as the value used for Ec. Overall, a customized density-modulus relationship for the ulna was generated, which provided improved results over using previously reported density-modulus equations.

Quantifying lung morphology with respiratory-gated micro-CT in a murine model of emphysema.

Non-invasive micro-CT imaging techniques have been developed to investigate lung structure in free-breathing rodents. In this study, we investigate the utility of retrospectively respiratory-gated micro-CT imaging in an emphysema model to determine if anatomical changes could be observed in the image-derived quantitative analysis at two respiratory phases. The emphysema model chosen was a well-characterized, genetically altered model (TIMP-3 knockout mice) that exhibits a homogeneous phenotype. Micro-CT scans of the free-breathing, anaesthetized mice were obtained in 50 s and retrospectively respiratory sorted and reconstructed, providing 3D images representing peak inspiration and end expiration with 0.15 mm isotropic voxel spacing. Anatomical measurements included the volume and CT density of the lungs and the volume of the major airways, along with the diameters of the trachea, left bronchus and right bronchus. From these measurements, functional parameters such as functional residual capacity and tidal volume were calculated. Significant differences between the wild-type and TIMP-3 knockout groups were observed for measurements of CT density over the entire lung, indicating increased air content in the lungs of TIMP-3 knockout mice. These results demonstrate retrospective respiratory-gated micro-CT, providing images at multiple respiratory phases that can be analyzed quantitatively to investigate anatomical changes in murine models of emphysema.

Use of co-registered high-resolution computed tomography scans before and after screw insertion as a novel technique for bone mineral density determination along screw trajectory.

INTRODUCTION: Bone mineral density (BMD) is an important factor in the examination of the performance of bone instrumentation both in and ex vivo, and until now, there has not existed a reliable technique for determining BMD at the precise location of such hardware. This paper describes such a technique, using cadaveric human sacra as a model. METHODS: Nine fresh-frozen sacra had solid and hollow titanium screws placed into the S1 pedicles from a posterior approach. High-resolution micro-computed tomography (CT) was performed on each specimen before and after screw placement. All images were reconstructed with an isotropic spatial resolution of 308 mum, reoriented, and the pre-screw and post-screw scans were registered and transformed using a six-degree rigid-body transformation matrix. Once registered, two points, corresponding to the center of the screw at the cortex and at the screw tip, were determined in each scan. These points were used to generate cylindrical regions of interest (ROI) with the same trajectory and dimensions as the screw. BMD measurements were obtained within each of the ROI in the pre-screw scan. To examine the effect of artefact on BMD measurements around the titanium screws, annular ROI of 1 mm thickness were created expanding from the surface of the screws, and BMD was measured within each in both the pre- and post-screw scans. RESULTS: The registration process was accurate to 190 mum, with a precision of 189 mum and error in BMD measurement of +/-2% in repeated scans. BMD values in the cylindrical ROI corresponding to screw trajectories were not statistically different from side to side of each specimen (p=0.23). Metal artefact created significant differences in BMD values (p=0.001) and followed an exponential decay curve as distance from the screws increased, approaching a low value of approximately 20 mg HA cm(-3), but not disappearing completely. SUMMARY: CT in the presence of metal creates artefact, making measured BMD values near implants unreliable. This technique is accurate for determination of BMD, non-destructive, and eliminates the problem of this metal artefact through the use of co-registered scans. This technique has applications both in vitro and in vivo.

Implementation of dual- and triple-energy cone-beam micro-CT for postreconstruction material decomposition.

Micro-CT has become a powerful tool for small animal research, having the ability to obtain high-resolution in vivo and ex vivo images for analyzing bone mineral content, organ vasculature, and bone microarchitecture extraction. The use of exogenous contrast agents further extends the use of micro-CT techniques, but despite advancements in contrast agents, single-energy micro-CT is still limited in cases where two different materials share similar grey-scale intensity values. This study specifically addresses the development of multiple-energy cone-beam micro-CT, for applications where bone must be separated from blood vessels filled with a Pb-based contrast material (Microfil) in ex vivo studies of rodents and tissue specimens. The authors report the implementation of dual- and triple-energy CT algorithms for material-specific imaging using postreconstruction decomposition of micro-CT data; the algorithms were implemented on a volumetric cone-beam micro-CT scanner (GE Locus Ultra). For the dual-energy approach, extrinsic filtration was applied to the x-ray beam to produce spectra with different proportions of x rays above the K edge of Pb. The optimum x-ray tube energies (140 kVp filtered with 1.45 mm Cu and 96 kVp filtered with 0.3 mm Pb) that maximize the contrast between bone and Microfil were determined through numerical simulation. For the triple-energy decomposition, an additional low-energy spectrum (70 kVp, no added filtration) was used. The accuracy of decomposition was evaluated through simulations and experimental verification of a phantom containing a cortical bone simulating material (SB3), Microfil, and acrylic. Using simulations and phantom experiments, an accuracy greater than 95% was achieved in decompositions of bone and Microfil (for noise levels lower than 11 HU), while soft tissue was separated with accuracy better than 99%. The triple-energy technique demonstrated a slightly higher, but not significantly different, decomposition accuracy than the dual-energy technique for the same achieved noise level in the micro-CT images acquired at the multiple energies. The dual-energy technique was applied to the decomposition of an ex vivo rat specimen perfused with Microfil; successful decomposition of the bone and Microfil was achieved, enabling the visualization and characterization of the vasculature both in areas where the vessels traverse soft tissue and when they are surrounded by bone. In comparison, in single energy micro-CT, vessels surrounded by bone could not be distinguished from the cortical bone, based on grey-scale intensity alone. This work represents the first postreconstruction application of material-specific decomposition that directly takes advantage of the K edge characteristics of a contrast material injected into an animal specimen; the application of the technique resulted in automatic, accurate segmentation of 3D micro-CT images into bone, vessel, and tissue components. The algorithm uses only reconstructed images, rather than projection data, and is calibrated by an operator with signal values in regions identified as being comprised entirely of either cortical bone, contrast-enhanced vessel, or soft tissue; these required calibration values are observed directly within reconstructed CT images acquired at the multiple energies. These features facilitate future implementation on existing research micro-CT systems.

In vivo small-animal imaging using micro-CT and digital subtraction angiography.

Small-animal imaging has a critical role in phenotyping, drug discovery and in providing a basic understanding of mechanisms of disease. Translating imaging methods from humans to small animals is not an easy task. The purpose of this work is to review in vivo x-ray based small-animal imaging, with a focus on in vivo micro-computed tomography (micro-CT) and digital subtraction angiography (DSA). We present the principles, technologies, image quality parameters and types of applications. We show that both methods can be used not only to provide morphological, but also functional information, such as cardiac function estimation or perfusion. Compared to other modalities, x-ray based imaging is usually regarded as being able to provide higher throughput at lower cost and adequate resolution. The limitations are usually associated with the relatively poor contrast mechanisms and potential radiation damage due to ionizing radiation, although the use of contrast agents and careful design of studies can address these limitations. We hope that the information will effectively address how x-ray based imaging can be exploited for successful in vivo preclinical imaging.

Poster - Thurs Eve-12: A needle-positioning robot co-registered with volumetric x-ray micro-computed tomography images for minimally-invasive small-animal interventions.

Preclinical research protocols often require the delivery of biological substances to specific targets in small animal disease models. To target biologically relevant locations in mice accurately, the needle positioning error needs to be < 200 µm. If targeting is inaccurate, experimental results can be inconclusive or misleading. We have developed a robotic manipulator that is capable of positioning a needle with a mean error < 100 µm. An apparatus and method were developed for integrating the needle-positioning robot with volumetric micro-computed tomography image guidance for interventions in small animals. Accurate image-to-robot registration is critical for integration as it enables targets identified in the image to be mapped to physical coordinates inside the animal. Registration is accomplished by injecting barium sulphate into needle tracks as the robot withdraws the needle from target points in a tissue-mimicking phantom. Registration accuracy is therefore affected by the positioning error of the robot and is assessed by measuring the point-to-line fiducial and target registration errors (FRE, TRE). Centroid points along cross-sectional slices of the track are determined using region growing segmentation followed by application of a center-of-mass algorithm. The centerline points are registered to needle trajectories in robot coordinates by applying an iterative closest point algorithm between points and lines. Implementing this procedure with four fiducial needle tracks produced a point-to-line FRE and TRE of 246 ± 58 µm and 194 ± 18 µm, respectively. The proposed registration technique produced a TRE < 200 µm, in the presence of robot positioning error, meeting design specification.

Study of subchondral bone adaptations in a rodent surgical model of OA using in vivo micro-computed tomography.

OBJECTIVE: To non-invasively investigate the changes to epiphyseal bone occurring in a longitudinal pre-clinical model of osteoarthritis (OA) using in vivo micro-computed tomography (micro-CT). DESIGN: In vivo micro-CT images were acquired using a bench-top micro-CT scanner, which produces three-dimensional data with isotropic voxel spacing of 0.046 mm. Male rodents were scanned prior to surgical destabilization, consisting of anterior cruciate ligament transection and partial medial menisectomy (ACLX). Subsequent scans were performed every 4 weeks post-ACLX, for up to 5 months. Volumetric bone mineral density (vBMD) was measured in specific, anatomically segmented regions within each image. The ACLX rodent data were compared with the contralateral non-operated hind limb of the same animal, as well as a sham-operated group (SHAM) of animals, for each time point. End-point histology compared changes to cartilage and bone between the ACLX and control animals. RESULTS: The micro-CT protocol produced sufficient spatial resolution and signal-to-noise ratio (SNR=19) to quantify subchondral bone pathology, with an acceptable entrance exposure to radiation (0.36 Gy). Significantly lower vBMD was measured in the ACLX group, vs SHAM rodents, at 1, 4, and 5 months post-surgery (P<0.05). Qualitative observations of ACLX joints revealed significant loss of cartilage, subchondral bone cysts, and calcification of tendon similar to changes found in humans. CONCLUSIONS: This study demonstrates in vivo micro-CT as an effective method for investigating the development of rodent knee OA longitudinally. This method can be applied, in future pre-clinical trials, to non-destructively monitor the efficacy of pharmacological interventions.

In vivo characterization of lung morphology and function in anesthetized free-breathing mice using micro-computed tomography.

Lung morphology and function in human subjects can be monitored with computed tomography (CT). Because many human respiratory diseases are routinely modeled in rodents, a means of monitoring the changes in the structure and function of the rodent lung is desired. High-resolution images of the rodent lung can be attained with specialized micro-CT equipment, which provides a means of monitoring rodent models of lung disease noninvasively with a clinically relevant method. Previous studies have shown respiratory-gated images of intubated and respirated mice. Although the image quality and resolution are sufficient in these studies to make quantitative measurements, these measurements of lung structure will depend on the settings of the ventilator and not on the respiratory mechanics of the individual animals. In addition, intubation and ventilation can have unnatural effects on the respiratory dynamics of the animal, because the airway pressure, tidal volume, and respiratory rate are selected by the operator. In these experiments, important information about the symptoms of the respiratory disease being studied may be missed because the respiration is forced to conform to the ventilator settings. In this study, we implement a method of respiratory-gated micro-CT for use with anesthetized free-breathing rodents. From the micro-CT images, quantitative analysis of the structure of the lungs of healthy unconscious mice was performed to obtain airway diameters, lung and airway volumes, and CT densities at end expiration and during inspiration. Because the animals were free breathing, we were able to calculate tidal volume (0.09 +/- 0.03 ml) and functional residual capacity (0.16 +/- 0.03 ml).

4 T MRI of chondrocalcinosis in combination with three-dimensional CT, radiography, and arthroscopy: a report of three cases.

OBJECTIVE: To describe 4 T MRI techniques in imaging chondrocalcinosis within the knee and examine the results together with those demonstrated using three-dimensional (3D) computed tomography, conventional radiography, and arthroscopy. DESIGN AND PATIENTS: From a larger clinical imaging study of early osteoarthritis, knee arthroscopy patients were imaged using high-field MRI and high-resolution 3D CT prior to their surgery. Retrospective review of the imaging data diagnosed three patients with chondrocalcinosis. Fat-suppressed 3D spoiled gradient (3D SPGR) and two-dimensional fat-suppressed fast spin echo (FSE) imaging was performed at 4 T. The MR images, multi-planar reformatted CT (MPR-CT) and maximum intensity projection CT (MIP-CT) images, and radiographs were examined by a musculoskeletal radiologist for the presence and location of chondrocalcinosis. The findings from arthroscopy were also included. RESULTS: MRI showed 16 sites of punctate hypointense regions from 18 articular surfaces and five of six menisci with similar signal characteristics. Both meniscal chondrocalcinosis and meniscal tears were clearly visible using the 3D SPGR sequence. Only three sites were demonstrated to have calcification using MPR-CT and MIP-CT revealed an additional three. In articular cartilage surfaces showing surface disruption, arthroscopy demonstrated 11 sites with crystal deposition. Arthroscopy also revealed five menisci with calcification present. CONCLUSION: Our preliminary findings suggest that imaging chondrocalcinosis using spoiled gradient 4 T MRI is superior and complementary to the other imaging modalities in the detection of crystal deposition in both articular cartilage and menisci.

Fundamental image quality limits for microcomputed tomography in small animals.

Small-animal imaging has become increasingly more important as transgenic and knockout mice are produced to model human diseases. One imaging technique that has emerged is microcomputed tomography (micro-CT). For live-animal imaging, the precision in the images will be determined by the x-ray dose given to the animal. As a result, we propose a simple method to predict the noise performance of an x-ray micro-CT system as a function of dose and image resolution. An ideal, quantum-noise limited micro-CT scanner, assumed to have perfect resolution and ideal efficiency, was modeled. Using a simplified model, the coefficient of variation (COV) of the linear attenuation coefficient was calculated for a range of entrance doses and isotropic voxel sizes. COV calculations were performed for the ideal case and with simulated imperfections in efficiency and resolution. Our model was validated in phantom studies and mouse images were acquired with a specimen scanner to illustrate the results. A simplified model of noise propagation in the case of isotropic resolution indicates that the COV in the linear attenuation coefficient is proportional to (dose)(-1/2) and to the (isotropic voxel size)(-2) in the reconstructed volume. Therefore an improvement in the precision can be achieved only by increasing the isotropic voxel size (thereby decreasing the resolution of the image) or by increasing the x-ray dose. For the ideal scanner, a COV of 1% in the linear attenuation coefficient for an image of a mouse exposed to 0.25 Gy is obtained with a minimum isotropic voxel size of 135 microm. However, the same COV is achieved at a dose of 5.0 Gy with a 65 microm isotropic voxel size. Conversely, for a 68 mm diameter rat, a COV of 1% obtained from an image at 5.0 Gy would require an isotropic voxel size of 100 microm. These results indicate that short-term, potentially lethal, effects of ionizing radiation will limit high-resolution live animal imaging. As improvements in detector technology allow the resolution to improve, by decreasing the detector element size to tens of microns or less, high quality images will be limited by the x-ray dose administered. For the highest quality images, these doses will approach the lethal dose or LD50 for the animals. Approaching the lethal dose will affect the way experiments are planned, and may reduce opportunities for experiments involving imaging the same animal over time. Dose considerations will become much more important for live small-animal imaging as the limits of resolution are tested.