BackgroundTherapy with mesenchymal stem cells (MSCs) has been reported to provide beneficial effects in the treatment of neurological and orthopaedic disorders in dogs. The exact mechanism of action is poorly understood. Magnetic resonance imaging (MRI) gives the opportunity to observe MSCs after clinical administration. To visualise MSCs with the help of MRI, labelling with an MRI contrast agent is necessary. However, it must be clarified whether there is any negative influence on cell function and viability after labelling prior to clinical administration.ResultsFor the purpose of the study, seven samples with canine adipose-derived stem cells were incubated with superparamagnetic iron oxide nanoparticles (SPIO: 319.2 μg/mL Fe) for 24 h. The internalisation of the iron particles occurred via endocytosis. SPIO particles were localized as free clusters in the cytoplasm or within lysosomes depending on the time of investigation. The efficiency of the labelling was investigated using Prussian blue staining and MACS assay. After 3 weeks the percentage of SPIO labelled canine stem cells decreased. Phalloidin staining showed no negative effect on the cytoskeleton. Labelled cells underwent osteogenic and adipogenic differentiation. Chondrogenic differentiation occurred to a lesser extent compared with a control sample. MTT-Test and wound healing assay showed no influence of labelling on the proliferation. The duration of SPIO labelling was assessed using a 1 Tesla clinical MRI scanner and T2 weighted turbo spin echo and T2 weighted gradient echo MRI sequences 1, 2 and 3 weeks after labelling. The hypointensity caused by SPIO lasted for 3 weeks in both sequences.ConclusionsAn Endorem labelling concentration of 319.2 μg/mL Fe (448 μg/mL SPIO) had no adverse effects on the viability of canine ASCs. Therefore, this contrast agent could be used as a model for iron oxide labelling agents. However, the tracking ability in vivo has to be evaluated in further studies.
Sonography is a convenient, non-invasive diagnostic modality in small ruminants, often used in reproductive management, internal medicine, and surgery. Pregnancy diagnostics and imaging anatomy and pathology of organs, such as neoplasia, are major applications. Urolithiasis is one of the most common causes of death in male sheep and goats, for which sonography is the diagnostic modality of choice. Although ultrasound-imaging techniques for kidneys, ureters, and urinary bladder in small ruminants have been described previously, this study focuses on reporting the imaging technique of the extra-pelvic portion of the urethra, as nearly all the cases of obstructive urolithiasis result from urinary stones in this part. Thirty-three Lacaune-crossbred lambs were examined using a 12 MHz linear probe in laterally recumbent animals. Using this technique, the urethral lumen could be visualised through its entire course in all lambs.
Objectives The aim of this study was to determine the radiographic sensitivity in detecting medial coronoid disease (MCD), using computed tomography (CT) as reference in dogs presented for an official second opinion, and to compare the medial coronoid process (MCP) in fragmented and fissured MCP as well as those unaffected by MCD. Materials and Methods The data of dogs, presented for official second opinion radiographs and CT, were reviewed by three board-certified observers and in accordance with the International Elbow Working Group guidelines regarding MCD. Radiographic delineation, radiopacity and Hounsfield Units (HU) of the MCP were recorded additionally and the correlation between radiography and CT was investigated. Results Sensitivity and specificity of radiography compared with CT yielded values of 83.6% for the former and 83.5% for the latter. False-negative grading in radiography correlated significantly (p = 0.0001) with a present fissure line in CT. The mean delineation (p = 0.03) and mean HU of fragmented MCP (p = 0.0045) were significantly reduced compared with fissured MCP and no significant differences in measured HU for fissured MCP were detected in comparison to elbows unaffected by MCD. Conclusion The results of the present study show substantial agreement between radiography and CT in second opinion cases. However, sensitivity is reduced compared with not preselected cases. A present fissure line in CT was significantly associated with a false-negative grading in radiography. Therefore, CT imaging of the elbows is strongly recommended in cases of appeal.
Bovine tuberculosis (bTB) not only poses a zoonotic threat to humans but also has a significant economic impact on livestock production in many areas of the world. Effective vaccines for humans, livestock, and wildlife are highly desirable to control tuberculosis. Suitable large animal models are indispensable for meaningful assessment of vaccine candidates. Here, we describe the refinement of an animal model for bTB in goats. Intrabronchial inoculation procedure via video-guided endoscopy in anesthetized animals, collection of lungs after intratracheal fixation in situ, and imaging of lungs by computed tomography (CT) were established in three goats using barium sulfate as surrogate inoculum. For subsequent infection experiments, four goats were infected with 4.7 × 102 colony-forming units of M. bovis by intrabronchial inoculation using video-guided endoscopy with spray catheters. Defined amounts of inoculum were deposited at five sites per lung. Four age-matched goats were mock-inoculated. None of the goats developed clinical signs until they were euthanized 5 months post infection, but simultaneous skin testing confirmed bTB infection in all goats inoculated with M. bovis. In tissues collected at necropsy, M. bovis was consistently re-isolated from granulomas in lymph nodes, draining the lungs of all the goats infected with M. bovis. Further dissemination was observed in one goat only. Pulmonary lesions were quantified by CT and digital 2D radiography (DR). CT revealed mineralized lesions in all the infected goats ranging from <5 mm to >10 mm in diameter. Small lesions <5 mm predominated. The DR failed to detect small lesions and to determine the exact location of lesions because of overlapping of pulmonary lobes. Relative volume of pulmonary lesions was low in three but high in one goat that also had extensive cavitation. CT lesions could be correlated to gross pathologic findings and histologic granuloma types in representative pulmonary lobes. In conclusion, video-guided intrabronchial inoculation with spray catheters, mimicking the natural way of infection, resulted in pulmonary infection of goats with M. bovis. CT, but not DR, presented as a highly sensitive method to quantify the extent of pulmonary lesions. This goat model of TB may serve as a model for testing TB vaccine efficacy.
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