Comparison of high-resolution X-ray and micro-CT for experimental evaluation of intracranial stent prototypes: quality evaluation beyond CE mark.

INTRODUCTION: As digital mammography and micro-computed tomography (CT) have been used for evaluation of stents deployed in experimental animal models, we compared the two methods regarding their sensitivity to detect abnormalities in three prototypes of intracranial stents. METHODS: Three different prototypes of intracranial stents (n = 84) were implanted in various animal models. Explanted stents were examined using digital mammography and micro-CT. The images were compared with respect to maintenance of material and form and the stents were compared to one another. Histological analysis was performed as well. RESULTS: In the open-cell stents, expansion of the stent cells was detected in the majority of cases (57.1 %) using micro-CT and less frequently using mammography (42.3 %). The closed-cell stent revealed kink stenoses in mammography as well as in micro-CT (3/7, 42.9 %). Detailed reconstructions of micro-CT images showed high-grade kink stenoses of the flow-diverter stent in two extremely curved vessels. Strut breaks were observed more frequently using micro-CT (6/84, 7.1 %) than by mammography (4/84, 4.8 %). Histology confirmed all changes of stent architecture. CONCLUSION: Significant changes of stent architecture can be observed and assessed even in the two-dimensional mammographic images. The use of micro-CT is recommended to detect subtle changes like single strut breaks and for three-dimensional information.

Characterization and longitudinal monitoring of melanoma growth in ret-transgenic mice using a single-sequence MRI protocol.

Spontaneous melanoma models in transgenic mice are increasingly used in preclinical research as they most closely match the progression of melanoma in humans. While optical inspection only allows analysis of tumors located on the skin, the accurate measurement and growth of subcutaneous tumors have not been adequately assessed. To improve the measurement accuracy of melanoma tumors, we used a fast single-sequence MRI protocol at 9.4 Tesla for longitudinal characterization of a ret-transgenic mouse model. Repeated MRI (average acquisition time 30 min per animal) of the trunk (excluding head and distal limbs) in six siblings revealed an increase in the mean total tumor volume (TTV) from 102.0 ± 80.5 mm(3) at 35 days of age to 434.8 ± 154.9 mm(3) by 77 days. The main tumor load was located within the pelvis (>40%), followed by the proximal hind limbs and groins (>30%). The smallest detectable tumor measured 0.07 mm(3). Inter-rater reliability between a radiologist and a veterinarian analysing MRI data was 0.993 for TTV and 0.840 for number of tumors (both p < 0.001). We thus conclude that because of the high variance of TTV of same-aged mice, MRI should be used (i) to establish treatment groups matched for TTV and (ii) for longitudinal examination of the TTV in mice over the course of treatments.

Double-contrast micro-CT colonoscopy in live mice.

PURPOSE: Models of colon cancer in small rodents are of particular interest as they most closely simulate the development and growth of colonic cancer in humans. Micro-computed tomography has been used for detection of polyps in murine models of colon cancer. The study was performed to evaluate whether a novel high-speed continuous-rotation, single-breath-hold scanning protocol in combination with double-contrasting of the colon can be successfully applied for colonoscopy of live mice at acquisition times of 40 s. METHODS: C57BL/6JApcMin/+ mice were intubated and ventilated. After double-contrasting the colon with barium and air, mice underwent continuous rotation micro-CT (mean resolution 41 × 41 × 53 µm) during a single-breath-hold period of 40 s. Sensitivity to detect colon polyps by four blinded radiologists was analysed. Number and location of polyps were verified in the excised colon. Radiation dose was measured using a thermoluminescence dosimeter placed within the distal colon. RESULTS: In six of seven mice, a total of 12 polyps were detected in the explanted colon (one mouse without polyps). One tumor (8.3%) was located in the proximal third, seven tumors (58.1%) and four tumors (33.2%) were located in the middle and in the distal third of the colon, respectively. Mean tumor volume was 6.5 ± 3.6 mm(3). Sensitivity to detect colon polyps was 0.85 ± 0.1. Mean radiation dose was 0.241 ± 0.002 Gy. CONCLUSION: Using a high-speed continuous rotation micro-CT protocol, double-contrast single-breath-hold colonoscopy in mice is feasible and yields sufficient contrast to visualize the proximal colonic folds and to detect colonic polyps in vivo.

Evaluation of a continuous-rotation, high-speed scanning protocol for micro-computed tomography.

OBJECTIVE: Micro-computed tomography is used frequently in preclinical in vivo research. Limiting factors are radiation dose and long scan times. The purpose of the study was to compare a standard step-and-shoot to a continuous-rotation, high-speed scanning protocol. METHODS: Micro-computed tomography of a lead grid phantom and a rat femur was performed using a step-and-shoot and a continuous-rotation protocol. Detail discriminability and image quality were assessed by 3 radiologists. The signal-to-noise ratio and the modulation transfer function were calculated, and volumetric analyses of the femur were performed. The radiation dose of the scan protocols was measured using thermoluminescence dosimeters. RESULTS: The 40-second continuous-rotation protocol allowed a detail discriminability comparable to the step-and-shoot protocol at significantly lower radiation doses. No marked differences in volumetric or qualitative analyses were observed. CONCLUSIONS: Continuous-rotation micro-computed tomography significantly reduces scanning time and radiation dose without relevantly reducing image quality compared with a normal step-and-shoot protocol.

High-speed single-breath-hold micro-computed tomography of thoracic and abdominal structures in mice using a simplified method for intubation.

OBJECTIVES: Respiratory gating with and without controlled ventilation has been applied for in vivo micro-computed tomography (micro-CT) of thoracic and abdominal structures in mice. We describe a simplified method for intubation and demonstrate its applicability for single-breath-hold micro-CT in mice. METHODS: Mice (n = 10) were anesthetized, intubated, ventilated, and relaxed by intraperitoneal administration of rocuronium. Contrast-enhanced micro-CT of the complete thorax including the upper abdominal organs (80 kV; 37.5 µA; 190-degree rotation; 600 projections/20 seconds or 1200 projections/40 seconds; 39 × 39 × 50-µm voxel size) was performed with and without single-breath-hold technique. RESULTS: The simplified method of intubation was fast (<1 minute) and required no special hardware in all mice. Relaxation of mice allowed prolonged single-breath-hold imaging of up to 40 seconds. Diameter of smallest identifiable lung vessels was 100 µm. CONCLUSIONS: The presented simplified method for intubation in mice is fast, safe, and effective. Additional relaxation allowed high-resolution single-breath-hold micro-CT in mice.

Radioimmunoimaging of liver metastases with PET using a 64Cu-labeled CEA antibody in transgenic mice.

PURPOSE: Colorectal cancer is one of the most common forms of cancer, and the development of novel tools for detection and efficient treatment of metastases is needed. One promising approach is the use of radiolabeled antibodies for positron emission tomography (PET) imaging and radioimmunotherapy. Since carcinoembryonic antigen (CEA) is an important target in colorectal cancer, the CEA-specific M5A antibody has been extensively studied in subcutaneous xenograft models; however, the M5A antibody has not yet been tested in advanced models of liver metastases. The aim of this study was to investigate the (64)Cu-DOTA-labeled M5A antibody using PET in mice bearing CEA-positive liver metastases. PROCEDURES: Mice were injected intrasplenically with CEA-positive C15A.3 or CEA-negative MC38 cells and underwent micro-computed tomography (micro-CT) to monitor the development of liver metastases. After metastases were detected, PET/MRI scans were performed with (64)Cu-DOTA-labeled M5A antibodies. H&E staining, immunohistology, and autoradiography were performed to confirm the micro-CT and PET/MRI findings. RESULTS: PET/MRI showed that M5A uptake was highest in CEA-positive metastases. The %ID/cm(3) (16.5% ± 6.3%) was significantly increased compared to healthy liver tissue (8.6% ± 0.9%) and to CEA-negative metastases (5.5% ± 0.6%). The tumor-to-liver ratio of C15A.3 metastases and healthy liver tissue was 1.9 ± 0.7. Autoradiography and immunostaining confirmed the micro-CT and PET/MRI findings. CONCLUSION: We show here that the (64)Cu-DOTA-labeled M5A antibody imaged by PET can detect CEA positive liver metastases and is therefore a potential tool for staging cancer, stratifying the patients or radioimmunotherapy.

Implantation of pipeline flow-diverting stents reduces aneurysm inflow without relevantly affecting static intra-aneurysmal pressure.

BACKGROUND: Flow-diverting stent (FDS) implantation is an endovascular treatment option for intracranial aneurysms. However, little is known about the hemodynamic effects. OBJECTIVE: To assess the effect of stent compression on FDS porosity, to evaluate the influence of single and overlapping implantation of FDS on intra-aneurysmal flow profiles, and to correlate stent porosity with changes in static mean intra-aneurysmal pressure. METHODS: Intra-aneurysmal time-density curves were recorded in a pulsatile in vitro flow model before and after implantation of FDSs (Pipeline Embolization Device; ev3) in 7 different types of aneurysm models. Reductions in the maximum contrast inflow and time to maximum intra-aneurysmal contrast were calculated. Micro--computed tomography was performed, and compression-related FDS porosity was measured. The influence of FDS placement on mean static intra-aneurysmal pressure was measured. RESULTS: FDS compression resulted in an almost linear reduction in stent porosity. Stent porosity (struts per 1 mm) correlated significantly with the reduction of aneurysm contrast inflow (R = 0.81, P < .001) and delay until maximum contrast (R = 0.34, P = .001). Circulating intra-aneurysmal high-velocity flow was terminated in all sidewall models after implantation of a single stent. Superimposition of 2 stents reduced maximum intra-aneurysmal contrast by 69.1 ± 3.1% (mean ± SD) in narrow-necked sidewall aneurysm models, whereas no substantial reduction in maximum intra-aneurysmal contrast was observed in wide-necked sidewall aneurysm models. Intra-aneurysmal mean static pressure did not correlate with FDS porosity or number of implanted stents. CONCLUSION: Implantation of FDS effectively reduces aneurysm inflow in a porosity-dependent way without relevantly affecting static mean intra-aneurysmal pressure. ABBREVIATIONS: FDS, flow-diverting stentMAP, mean arterial pressurePED, Pipeline Embolization Device.

Comparison of Fenestra LC, ExiTron nano 6000, and ExiTron nano 12000 for micro-CT imaging of liver and spleen in mice.

RATIONALE AND OBJECTIVES: The purpose of this study was to compare different contrast agents for longitudinal liver and spleen imaging in a mouse model of liver metastasis. MATERIALS AND METHODS: Mice developing liver metastases underwent longitudinal micro-computed tomography imaging after injection of Fenestra LC, ExiTron nano 6000, or ExiTron nano 12000. Elimination times and contrast enhancement of liver and spleen were compared. RESULTS: For all contrast agents, liver contrast peaked at approximately 4 hours and spleen contrast at 48 hours postinjection. A single dose of 100 µL of ExiTron nano 6000 or 12000 resulted in longstanding enhancement of liver and spleen tissue for longer than 3 weeks, whereas repeated injections of 400 µL of Fenestra LC were required to retain contrast at acceptable levels and allowed imaging of the liver/spleen for up to 2 and 9 days, respectively. CONCLUSION: Both ExiTron nano agents provide longer and stronger contrast enhancement of liver and spleen compared to Fenestra LC, and they do so at a 75% lower injection volume in mice.

A Low Cost Metal-Free Vascular Access Mini-Port for Artifact Free Imaging and Repeated Injections in Mice.

PURPOSE: Small injection ports for mice are increasingly used for drug testing or when administering contrast agents. Commercially available mini-ports are expensive single-use items that cause imaging-artifacts. We developed and tested an artifact-free, low-cost, vascular access mini-port (VAMP) for mice. PROCEDURES: Leakage testing of the VAMP was conducted with high speed bolus injections of different contrast agents. VAMP-induced artifacts were assessed using a micro-CT and a small animal MRI (9.4T) scanner ex vivo. Repeated contrast administration was performed in vivo. RESULTS: With the VAMP there was no evidence of leakage with repeated punctures, high speed bolus contrast injections, and drawing of blood samples. In contrast to the tested commercially available ports, the VAMP did not cause artifacts with MRI or CT imaging. CONCLUSIONS: The VAMP is an alternative to commercially available mini-ports and has useful applications in animal research involving imaging procedures and contrast agent testing.

Comparison of digital subtraction angiography, micro-computed tomography angiography and magnetic resonance angiography in the assessment of the cerebrovascular system in live mice.

PURPOSE: Mice are often used as small animal models of brain ischemia, venous thrombosis, or vasospasm. This article aimed at providing an overview of the currently available methodologies for in vivo imaging of the murine cerebrovasculature and comparing the capabilities and limitations of the different methods. METHODS: Micro-computed tomography angiography (CTA) was performed during intra-arterial and intravenous administration of a contrast agent bolus. Digital subtraction angiography (DSA) was performed during intra-arterial administration of contrast agent using the micro-CT scanner. Time-of-flight (ToF) magnetic resonance (MR) angiography was performed using a small animal scanner (9.4 T) equipped with a cryogenic transceive quadrature coil. Datasets were compared for scan time, contrast-to-noise ratio (CNR), temporal and spatial resolution, radiation dose, contrast agent dose and detailed recognition of cerebrovascular structures. RESULTS: Highest spatial resolution was achieved using micro-CTA (16 x 16 x 16 µm) and DSA (14 x 14 µm). Compared to micro-CTA (20-40 s) and ToF-MRA (57 min), DSA provided highest temporal resolutions (30 fps) allowing analyses of the cerebrovascular blood flow. Highest mean CNR was reached using ToF-MRA (50.7 ± 15.0), while CNR of micro-CTA depended on the intra-arterial (19.0 ± 1.0) and intravenous (1.3 ± 0.4) use of agents. The CNR of DSA was 10.0 ± 1.8. CONCLUSIONS: The use of dedicated small animal scanners allows cerebrovascular imaging in live animals as small as mice. As each of the methods analyzed has its advantages and limitations, choosing the best suited imaging modality for a defined question is of great importance. By this means the aforementioned methods offer a great potential for future projects in preclinical cerebrovascular research including ischemic stroke or vasospasm.

Three-dimensional in vivo imaging of the murine liver: a micro-computed tomography-based anatomical study.

Various murine models are currently used to study acute and chronic pathological processes of the liver, and the efficacy of novel therapeutic regimens. The increasing availability of high-resolution small animal imaging modalities presents researchers with the opportunity to precisely identify and describe pathological processes of the liver. To meet the demands, the objective of this study was to provide a three-dimensional illustration of the macroscopic anatomical location of the murine liver lobes and hepatic vessels using small animal imaging modalities. We analysed micro-CT images of the murine liver by integrating additional information from the published literature to develop comprehensive illustrations of the macroscopic anatomical features of the murine liver and hepatic vasculature. As a result, we provide updated three-dimensional illustrations of the macroscopic anatomy of the murine liver and hepatic vessels using micro-CT. The information presented here provides researchers working in the field of experimental liver disease with a comprehensive, easily accessable overview of the macroscopic anatomy of the murine liver.

Micro-CT based experimental liver imaging using a nanoparticulate contrast agent: a longitudinal study in mice.

BACKGROUND: Micro-CT imaging of liver disease in mice relies on high soft tissue contrast to detect small lesions like liver metastases. Purpose of this study was to characterize the localization and time course of contrast enhancement of a nanoparticular alkaline earth metal-based contrast agent (VISCOVER ExiTron nano) developed for small animal liver CT imaging. METHODOLOGY: ExiTron nano 6000 and ExiTron nano 12000, formulated for liver/spleen imaging and angiography, respectively, were intravenously injected in C57BL/6J-mice. The distribution and time course of contrast enhancement were analysed by repeated micro-CT up to 6 months. Finally, mice developing liver metastases after intrasplenic injection of colon carcinoma cells underwent longitudinal micro-CT imaging after a single injection of ExiTron nano. PRINCIPAL FINDINGS: After a single injection of ExiTron nano the contrast of liver and spleen peaked after 4-8 hours, lasted up to several months and was tolerated well by all mice. In addition, strong contrast enhancement of abdominal and mediastinal lymph nodes and the adrenal glands was observed. Within the first two hours after injection, particularly ExiTron nano 12000 provided pronounced contrast for imaging of vascular structures. ExiTron nano facilitated detection of liver metastases and provided sufficient contrast for longitudinal observation of tumor development over weeks. CONCLUSIONS: The nanoparticulate contrast agents ExiTron nano 6000 and 12000 provide strong contrast of the liver, spleen, lymph nodes and adrenal glands up to weeks, hereby allowing longitudinal monitoring of pathological processes of these organs in small animals, with ExiTron nano 12000 being particularly optimized for angiography due to its very high initial vessel contrast.