Joy Dunmore-Buyze

A real vessel phantom for flow imaging: 3-d doppler ultrasound of steady flow

Vascular phantoms are used to assess the capabilities of various imaging techniques, such as x-ray CT and angiography, and B-mode, power Doppler, and colour Doppler ultrasound (US). They should, therefore, accurately mimic the vasculature, blood, and surrounding tissue, in regard to both imaging properties and vessel geometry. In the past, a variety of walled and wall-less vessel models have been used. However, these models only approximate the true vessel geometry, and generally lack pathologic features such as plaques or calcifications. To amend these deficiencies, we have developed a real vessel phantom for US and x-ray studies, which comprises a fixed human vessel specimen, cannulated onto two acrylic tubes, and embedded in agar in an acrylic box. Earlier, we demonstrated a good overall correlation between x-ray angiography, CT, and 3-D B-mode US images of this phantom. Here, we extend its use to flow imaging with 3-D power and 3-D colour Doppler US.

X-ray dose delivered during a longitudinal micro-CT study has no adverse effect on cardiac and pulmonary tissue in C57BL/6 mice

Background Micro-computed tomography (micro-CT) offers numerous advantages for small animal imaging, including the ability to monitor the same animals throughout a longitudinal study. However, concerns are often raised regarding the effects of X-ray dose accumulated over the course of the experiment. Purpose To scan C57BL/6 mice multiple times per week for 6 weeks, in order to determine the effect of the cumulative dose on pulmonary and cardiac tissue at the end of the study. Material and Methods C57BL/6 male mice were split into two groups (irradiated group = 10, control group = 10). The irradiated group was scanned (80 kVp/50mA) three times weekly for 6 weeks, resulting in a weekly dose of 0.84 Gy, and a total study dose of 5.04 Gy. The control group was scanned on the final week. Scans from week 6 were reconstructed and the lungs and heart were analyzed. Results Overall, there was no significant difference in lung volume or lung density or in left ventricular volume or ejection fraction between the control group and the irradiated group. Histological samples taken from excised lung and myocardial tissue also showed no evidence of inflammation or fibrosis in the irradiated group. Conclusion This study demonstrated that a 5 Gy X-ray dose accumulated over 6 weeks during a longitudinal micro-CT study had no significant effects on the pulmonary and myocardial tissue of C57BL/6 mice. As a result, the many advantages of micro-CT imaging, including rapid acquisition of high-resolution, isotropic images in free-breathing mice, can be taken advantage of in longitudinal studies without concern for negative dose-related effects.

Three-dimensional visualization and thickness estimation of aortic valve cusps using high-frequency ultrasound

High-frequency ultrasound techniques are introduced for three-dimensional imaging and thickness estimation of fresh heart valve cusps. Images of porcine aortic valve specimens were acquired within a 12 x 8 x 8 mm3 volume using a VisualSonics VS40 micro-imaging system operating at a 40 MHz centre frequency. Two image volumes were obtained from each of six left coronary cusps. One volume was acquired with the specimen submerged in distilled water and the second volume was acquired through either Hanks physiologic solution or coronary perfusion solution (CPS). The fibrosa, spongiosa and ventricularis were most readily distinguished when the specimen was imaged in distilled water. Colour thickness maps were computed from B-mode image data, and the mean and standard deviations of the thickness were determined for each cusp. In 11 of 12 trials, the image analysis algorithm yielded valid thickness estimates over greater than 98% of the region examined. Mean thickness estimates obtained with specimens submerged in Hanks solution or CPS ranged from 0.66 to 1.03 mm, and submersion in distilled water increased the mean thickness by 20-40%. This observation suggests that the cusps osmotically absorbed water. Information provided by high-frequency ultrasound is expected be valuable for characterizing the morphological properties of heart valves.

Time-course characterization of an aqueous colloidal polydisperse contrast agent in mice using micro-computed tomography

Background: Evaluation of cardiovascular function in mice using micro-CT requires that a contrast agent (CA) be administered to differentiate the blood from the myocardium. eXIA 160, an aqueous colloidal poly-disperse CA with a high concentration of iodine (160mg I/mL), creates strong contrast between blood and tissue with a low injection volume. In this study, the blood-pool enhancement time-course of eXIA 160 is monitored over a 24-hour period to determine its optimal use during cardiac function studies. Methods/Results: 8-second scans were performed (80kVp, 110mA) using the GE Locus Ultra micro-CT scanner. Male mice (black, 22-24g) were injected via tail vein with 5 μL/g body weight eXIA 160 (Binitio Biomedical Inc.). A precontrast scan was performed; following injection, mice were scanned at 15, 30, 45, and 60 minutes, 2, 4, 8, 24, and 48 hours. Overall, the highest contrast in the left ventricle occurred at 5 minutes (687 HU). Uptake of the CA by the myocardium was also observed: myocardial tissue showed increasing enhancement over a 4-hour period, remaining even once the contrast was eliminated from the vasculature. Conclusion: eXIA 160 provided high contrast between blood and myocardial tissue for a period of 30 minutes following injection. Notably, this CA was also taken up by the myocardium and provided continued enhancement when the contrast agent was eliminated from the blood, making LV wall function studies possible. In conclusion, eXIA 160, with its high iodine concentration and targeted tissue uptake characteristics, make it an ideal agent to use when evaluating cardiovascular function in mice.

Application of 3-d echocardiography and gated micro-computed tomography to assess cardiomyopathy in a mouse model of duchenne muscular dystrophy.

The purpose of this study was to measure changes in cardiac function as cardiomyopathy progresses in a mouse model of Duchenne muscular dystrophy using 3-D ECG-gated echocardiography. This study is the first to correlate cardiac volumes acquired using 3-D echocardiography with those acquired using retrospectively gated micro-computed tomography (CT). Both were further compared with standard M-mode echocardiography and histologic analyses. We found that although each modality measures a decrease in cardiac function as disease progresses in mdx/utrn(-/-) mice (n = 5) compared with healthy C57BL/6 mice (n = 8), 3-D echocardiography has higher agreement with gold-standard measurements acquired by gated micro-CT, with little standard deviation between measurements. M-Mode echocardiography measurements, in comparison, exhibit considerably greater variability and user bias. Given the radiation dose associated with micro-CT and the geometric assumptions made in M-mode echocardiography to calculate ventricular volume, we suggest that use of 3-D echocardiography has important advantages that may allow for the measurement of early disease changes that occur before overt cardiomyopathy.

Rapid microcomputed tomography suggests cardiac enlargement occurs during conductance catheter measurements in mice

Conductance catheters (CC) represent an established method of determining cardiac function in mice; however, the potentially detrimental effects a catheter may have on the mouse heart have never been evaluated. The present study takes advantage of rapid three-dimensional (3D) microcomputed tomography (CT) to compare simultaneously acquired micro-CT and CC measurements of left ventricular (LV) volumes in healthy and infarcted mice and to determine changes in LV volume and function associated with CC insertion. LV volumes were measured in C57BL/6 mice (10 healthy, 10 infarcted, 2% isoflurane anesthesia) using a 1.4-Fr Millar CC. 3D micro-CT images of each mouse were acquired before CC insertion as well as during catheterization. Each CT scan produced high-resolution images throughout the entire cardiac cycle in <1 min, enabling accurate volume measurements as well as direct visualization of the CC within the LV. Bland-Altman analysis demonstrated that CC measurements underestimate volume compared with CT measurements in both healthy [bias of -18.4 and -28.9 μl for end-systolic (ESV) and end-diastolic volume (EDV), respectively] and infarcted mice (ESV = -51.6 μl and EDV = -71.7 μl); underestimation was attributed to the off-center placement of the catheter. Individual evaluation of each heart revealed LV dilation following CC insertion in 40% of mice in each group. No change in ejection fraction was observed, suggesting the enlargement was caused by volume overload associated with disruption of the papillary muscles or chords. The enlargement witnessed was not significant; however, the results suggest the potential for CC insertion to detrimentally affect mouse myocardium, necessitating further investigation.

Vascularization of bioprosthetic valve material

Cell membrane remnants represent a probable nucleation site for calcium deposition in bioprosthetic heart valves. Calcification is a primary failure mode of both bovine pericardial and porcine aortic heterograft bioprosthesis but the nonuniform pattern of calcium distribution within the tissue remains unexplained. Searching for a likely cellular source, we considered the possibility of a previously overlooked small blood vessel network. Using a videomicroscopy technique, we examined 5 matched pairs of porcine aortic and pulmonary valves and 14 samples from 6 bovine pericardia. Tissue was placed on a Leitz Metallux microscope and transilluminated with a 75 watt mercury lamp. Video images were obtained using a silicon intensified target camera equipped with a 431 nm interference filter to maximize contrast of red cells trapped in a capillary microvasculature. Video images were recorded for analysis on a Silicon Graphics Image Analysis work station equipped with a video frame grabber. For porcine valves, the technique demonstrated a vascular bed in the central spongiosa at cusp bases with vessel sizes from 6-80 micrometers . Bovine pericardium differed with a more uniform distribution of 7-100 micrometers vessels residing centrally. Thus, small blood vessel endothelial cells provide a potential explanation patterns of bioprosthetic calcification.

Evaluating the dose effects of a longitudinal micro-CT study on pulmonary tissue in C57BL/6 mice

Background: Micro-computed tomography offers numerous advantages for small animal imaging, including the ability to monitor the same animals throughout a longitudinal study. However, concerns are often raised regarding the effects of x-ray dose accumulated over the course of the experiment. In this study, we scan C57BL/6 mice multiple times per week for six weeks, to determine the effect of the cumulative dose on pulmonary tissue at the end of the study. Methods/Results: C57BL/6 male mice were split into two groups (irradiated group=10, control group=10). The irradiated group was scanned (80kVp/50mA) each week for 6 weeks; the weekly scan session had three scans. This resulted in a weekly dose of 0.84 Gy, and a total study dose of 5.04 Gy. The control group was scanned on the final week. Scans from weeks 1 and 6 were reconstructed and analyzed: overall, there was no significant difference in lung volume or lung density between the control group and the irradiated group. Similarly, there were no significant differences between the week 1 and week 6 scans in the irradiated group. Histological samples taken from excised lung tissue also showed no evidence of inflammation or fibrosis in the irradiated group. Conclusion: This study demonstrates that a 5 Gy x-ray dose accumulated over six weeks during a longitudinal micro-CT study has no significant effects on the pulmonary tissue of C57BL/6 mice. As a result, the many advantages of micro- CT imaging, including rapid acquisition of high-resolution, isotropic images in free-breathing mice, can be taken advantage of in longitudinal studies without concern for negative dose-related effects.

Validation of high‐frequency ultrasound measurements of tissue layer thickness

High‐frequency ultrasound imaging enables nondestructive measurement of layer thickness in tissue specimens. These measurements are valuable for mechanical testing of soft biomaterials. This paper demonstrates a method for assessing the accuracy of high‐resolution ultrasonic thickness estimates. Three‐dimensional images of six porcine aortic valve cusps were acquired in vitro using a 40 MHz ultrasound system with 40×80×80 μm3 spatial resolution. The cusps were then frozen in liquid nitrogen, sectioned into 10‐μm slices, and micrographs of one slice from each specimen were acquired at 4× magnification. The two‐dimensional micrographs were registered to the three‐dimensional ultrasound images using a cross‐correlation method. The boundaries of the fibrosa, spongiosa, and ventricularis layers were segmented in both sets of images using an active contour model. The average thicknesses of the tissue layers in the registered images were estimated and the absolute differences of the optical and ultrasonic estima...