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Treatment of tendon disease with multipotent mesenchymal stromal cells (MSC) is a promising option to improve tissue regeneration. To elucidate the mechanisms by which MSC support regeneration, longitudinal tracking of MSC labelled with superparamagnetic iron oxide (SPIO) by magnetic resonance imaging (MRI) could provide important insight. Nine equine patients suffering from tendon disease were treated with SPIO-labelled or nonlabelled allogeneic umbilical cord-derived MSC by local injection. Labelling of MSC was confirmed by microscopy and MRI. All animals were subjected to clinical, ultrasonographical, and low-field MRI examinations before and directly after MSC application as well as 2, 4, and 8 weeks after MSC application. Hypointense artefacts with characteristically low signal intensity were identified at the site of injection of SPIO-MSC in T1- and T2∗-weighted gradient echo MRI sequences. They were visible in all 7 cases treated with SPIO-MSC directly after injection, but not in the control cases treated with nonlabelled MSC. Furthermore, hypointense artefacts remained traceable within the damaged tendon tissue during the whole follow-up period in 5 out of 7 cases. Tendon healing could be monitored at the same time. Clinical and ultrasonographical findings as well as T2-weighted MRI series indicated a gradual improvement of tendon function and structure.

Tendon disease has been treated with multipotent mesenchymal stromal cells (MSCs) in the equine large-animal model with promising success. The aim of this study was to gain more insight into the fate and biodistribution of MSCs after local application into tendon lesions by long-term cell tracking in this large-animal model. Superficial digital flexor tendon lesions were induced in all limbs in six horses and injected with 10106 Molday ION Rhodamine B-labeled MSCs suspended in serum or serum alone. Follow-up was performed using low-field magnetic resonance imaging (MRI), flow cytometry, and histology. Cell tracking based on the hypointense artifacts induced by the superparamagnetic iron oxide (SPIO) labeling agent in MRI as well as based on Rhodamine B fluorescence was feasible. However, Prussian blue staining for assessment of histology was not entirely specific for SPIO. Labeled cells could be traced at their injection site by MRI as well as histology for the whole follow-up period of 24 weeks. Although the numbers of labeled cells within the injected tendon lesions decreased over time, part of the applied cells appeared to remain viable and integrated within the injured tissue. Furthermore, small numbers of labeled cells were identified in peripheral blood within the first 24 h after cell injection and could also be found until week 24 within the contralateral control tendon lesions that had been injected with serum. The present findings unveil details on MSC biodistribution and persistence after their local application, which are of clinical relevance with regard to MSC safety and mechanisms of action.

SummaryReasons for performing study: Reductions in distances between dorsal spinous processes on radiographs are used as criteria for the diagnosis of impingement of the thoracic dorsal spinous processes in horses but are potentially altered by spine motion and different head and neck positions. Objectives: To determine the influence of head and neck positions on intervertebral distances between dorsal spinous processes on radiographs of thoracic spines of clinically sound horses. Methods: Lateral-lateral radiographs were obtained from 23 horses in 3 head and neck positions. The width of the thoracic dorsal spinous processes and intervertebral distances between adjacent thoracic dorsal spinous processes were measured at points perpendicular to a tangent between the dorsal spinous processes and the caudal extremity of the thoracic vertebrae. Results: A low head and neck position increased intervertebral distances between adjacent thoracic dorsal spinous processes from the 8th to 15th dorsal spinous processes whereas a high head and neck position had the opposite effect (P<0.05). Overall, intervertebral distances between adjacent thoracic dorsal spinous processes decreased from cranial to caudal in intermediate head and neck positions (P<0.01). The 12th thoracic dorsal spinous process was readily identifiable due to its significant difference to the narrower cranial and broader caudal dorsal spinous process (P<0.05). Conclusions:The head and neck position influences the distances between the dorsal spinous processes of the vertebrae of equine thoracic spine on radiography. Potential relevance: The measuring system reported here offers potential to improve and standardise radiographic evaluation of thoracic dorsal spinous processes.

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BackgroundMesenchymal stromal cells (MSC) have shown promising results in the treatment of tendinopathy in equine medicine, making this therapeutic approach seem favorable for translation to human medicine. Having demonstrated that MSC engraft within the tendon lesions after local injection in an equine model, we hypothesized that they would improve tendon healing superior to serum injection alone.MethodsQuadrilateral tendon lesions were induced in six horses by mechanical tissue disruption combined with collagenase application 3 weeks before treatment. Adipose-derived MSC suspended in serum or serum alone were then injected intralesionally. Clinical examinations, ultrasound and magnetic resonance imaging were performed over 24 weeks. Tendon biopsies for histological assessment were taken from the hindlimbs 3 weeks after treatment. Horses were sacrificed after 24 weeks and forelimb tendons were subjected to macroscopic and histological examination as well as analysis of musculoskeletal marker expression.ResultsTendons injected with MSC showed a transient increase in inflammation and lesion size, as indicated by clinical and imaging parameters between week 3 and 6 (p < 0.05). Thereafter, symptoms decreased in both groups and, except that in MSC-treated tendons, mean lesion signal intensity as seen in T2w magnetic resonance imaging and cellularity as seen in the histology (p < 0.05) were lower, no major differences could be found at week 24.ConclusionsThese data suggest that MSC have influenced the inflammatory reaction in a way not described in tendinopathy studies before. However, at the endpoint of the current study, 24 weeks after treatment, no distinct improvement was observed in MSC-treated tendons compared to the serum-injected controls. Future studies are necessary to elucidate whether and under which conditions MSC are beneficial for tendon healing before translation into human medicine.

Background Despite clinical importance and frequent occurrence of sinus disease, little is known about the size of paranasal sinuses and their communication in ponies and small horses. To examine the shape and volume of the paranasal sinuses and evaluate the sinonasal communication, three-dimensional (3D) reconstructions of computed tomography (CT) datasets of 12 healthy adult Shetland ponies were performed and analysed. Linear measurements of head length and width were taken. Using semi-automatic segmentation, 3D-models of all sinus compartments were created. Volumetric measurement of the seven sinus compartments were conducted and statistical analysis was performed. Sinus volumes were compared between the left and right sinuses and the relation to age and head size was evaluated. Results Structure and shape of the paranasal sinus system in Shetland ponies was similar to that of large horses. All seven sinus compartments on each side of the head were identified (rostral maxillary sinus, ventral conchal sinus, caudal maxillary sinus, dorsal conchal sinus, middle conchal sinus, frontal sinus, sphenopalatine sinus). The existence of a bilateral cranial and a caudal system formed by a maxillary septum was visible in all 12 individuals. The volumetric sizes of the left and right sinuses did not differ significantly (p > 0.05). A positive correlation between the size of the paranasal sinuses and the head length was shown. A relation between sinus volumes and age could not be proved in adult ponies aged > six years. Communication between single sinus compartments was identified. Furthermore, communication with the nasal cavity over the nasomaxillary aperture (Apertura nasomaxillaris) and a common sinonasal channel (Canalis sinunasalis communis) as well as its splitting up into a rostral and a caudolateral channel could be seen. Examination of the sinonasal communication was challenging and only a descriptive evaluation was possible. Conclusions Our findings concerning the size, shape and volumetric dimensions of Shetland pony CT images could help improve CT interpretation of abnormal clinical cases as well as aiding clinicians to develop and select appropriate instruments for medical inspection and treatments.

Scintigraphic examination of the thoracic spine is well documented. However, there is limited information about the effects of time on image quality in the period following injection of radionuclide. This study aimed to determine the optimal time point after injection of (99m)Tc-HDP (hydroxymethylene-diphosphonate) to achieve scintigraphic images with the best possible contrast and adequate count rates. Scintigraphic images of the thoracic spine of 21 horses were acquired two, four and six hours after administering (99m)Tc-HDP. Eight regions of interest were drawn in the images, four in the spinous processes and four in the adjacent soft tissue. The bone uptake, soft tissue uptake and the bone-to-soft tissue ratio were determined and compared between the different time points. Total count rates decreased with time after injection, but were at least as high as 150,000 counts per image at every time point after injection. The bone-to-soft tissue ratio was significantly higher for the images acquired after six hours compared to those acquired after two and four hours (P<0.01). Delayed scintigraphic examinations of the spinous processes of the equine thoracic spine achieved images with high contrast and sufficient count rates. Therefore, the scintigraphic examination of the equine thoracic spine is recommended to be done four to six hours after injecting (99m)Tc-HDP. However, additional studies should be performed to determine the effect of delayed image acquisition compared to images taken after three hours on the detectability of lesions in other parts of the thoracic spine and the soft tissue.