Human integrin α10ß1-selected mesenchymal stem cells home to cartilage defects in the rabbit knee and assume a chondrocyte-like phenotype.

BACKGROUND: Mesenchymal stem cells (MSCs) have shown promising results in stimulating cartilage repair and in the treatment of osteoarthritis (OA). However, the fate of the MSCs after intra-articular injection and their role in cartilage regeneration is not clear. To address these questions, this study investigated (1) homing of labeled human adipose tissue derived integrin α10ß1-selected MSCs (integrin α10-MSCs) to a cartilage defect in a rabbit model and (2) the ability of the integrin α10-MSCs to differentiate to chondrocytes and to produce cartilage matrix molecules in vivo. DESIGN: Integrin α10-MSCs were labeled with superparamagnetic iron oxide nanoparticles (SPIONs) co-conjugated with Rhodamine B to allow visualization by both MRI and fluorescence microscopy. A cartilage defect was created in the articular cartilage of the intertrochlear groove of the femur of rabbits. Seven days post-surgery, labeled integrin α10-MSCs or vehicle were injected into the joint. Migration and distribution of the SPION-labeled integrin α10-MSCs was evaluated by high-field 9.4 T MRI up to 10 days after injection. Tissue sections from the repair tissue in the defects were examined by fluorescence microscopy. RESULTS: In vitro characterization of the labeled integrin α10-MSCs demonstrated maintained viability, proliferation rate and trilineage differentiation capacity compared to unlabeled MSCs. In vivo MRI analysis detected the labeled integrin α10-MSCs in the cartilage defects at all time points from 12 h after injection until day 10 with a peak concentration between day 1 and 4 after injection. The labeled MSCs were also detected lining the synovial membrane at the early time points. Fluorescence analysis confirmed the presence of the labeled integrin α10-MSCs in all layers of the cartilage repair tissue and showed co-localization between the labeled cells and the specific cartilage molecules aggrecan and collagen type II indicating in vivo differentiation of the MSCs to chondrocyte-like cells. No adverse effects of the α10-MSC treatment were detected during the study period. CONCLUSION: Our results demonstrated migration and homing of human integrin α10ß1-selected MSCs to cartilage defects in the rabbit knee after intra-articular administration as well as chondrogenic differentiation of the MSCs in the regenerated cartilage tissue.

Identification and Quantification of Transient Receptor Potential Vanilloid 1 (TRPV1) in Equine Articular Tissue.

Joint pain and osteoarthritis (OA) are some of the most common causes of lameness in horses, and most of the available treatments focus on symptomatic relief without a disease-modifying effect. TRPV1 is a potential target for treating joint diseases, including OA, and the present study aims to investigate if the TRPV1 receptor is present in equine articular tissue and determine whether the number of receptors is upregulated in joint inflammation. Metacarpo/metatarsophalangeal (MCP/MTP) joints from 15 horses euthanised for reasons unrelated to this study were included. Based on synovial fluid analysis, macroscopic evaluation, and magnetic resonance imaging (MRI), joints were divided into two groups: healthy joints and joints with pathology. ELISA analysis was performed on synovial tissue harvested from all joints. TPRV1 was found in all joints. The mean concentration of TRPV1 compared to total protein in healthy joints (8.4 × 10-7 ng/mL) and joints with pathology (12.9 × 10-7 ng/mL) differed significantly (p = 0.01, t-test with Welch correction). Quantitative real-time reverse transcriptase PCR analysis was performed on RNA isolates from synovial tissue from all joints. TRPV1 mRNA expression ratio normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in healthy joints (0.16 (SD: 0.19)) and joints with pathology (0.24 (SD: 0.14)) did not differ significantly (p = 0.43, t-test with Welch correction). mRNA expression of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-) was very low for both groups. In conclusion, TRPV1 was detected both on mRNA and the protein level, with a higher expression of TRPV1 in samples from joints with pathology. Future studies will determine the clinical potential of equine TRPV1 as a target in the management of joint pain and inflammation.

Evaluation of gastrointestinal activity in healthy rabbits by means of duplex Doppler ultrasonography.

OBJECTIVE: To use duplex Doppler ultrasonography to compare gastrointestinal activity in healthy sedated versus nonsedated rabbits and to evaluate agreement between B-mode and pulsed-wave Doppler (PWD) ultrasonographic measurements. ANIMALS: 10 healthy client-owned rabbits brought for routine physical examination and 11 brought for routine ovariohysterectomy or castration. PROCEDURES: Duplex Doppler ultrasonography of the gastrointestinal tract was performed once for the 10 rabbits that underwent physical examination and twice (before and after presurgical sedation) for the 11 rabbits that underwent routine ovariohysterectomy or castration. Mean number of peristaltic contractions during a 30-second period was determined for the stomach, duodenum, jejunum, cecum, and colon from B-mode and PWD ultrasonographic images that had been video recorded. Findings for the duodenum and jejunum were compared between B-mode and PWD ultrasonography and between sedated and nonsedated rabbits. RESULTS: Duodenal and jejunal segments had measurable peristaltic waves; however, the stomach, cecum, and colon had no consistent measurable activity. B-mode and PWD ultrasonographic measurements for the duodenum and jejunum had high agreement. No significant difference was identified between nonsedated and sedated rabbits in mean number of peristaltic contractions of the duodenum or jejunum. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that both B-mode and PWD ultrasonography of the duodenum and jejunum may be suitable for noninvasive evaluation of small intestinal motility in rabbits and that the sedation protocol used in this study had no impact on measured peristaltic values.

Diagnosis of Soft Tissue Injury in the Sport Horse.

For successful diagnosis of soft tissue injuries in the sport horse, localizing the area of injury during clinical and lameness evaluation will be followed in most cases by an ultrasonographic examination. With MRI more available in equine veterinary clinics, this modality can allow for a complete evaluation of soft tissue and osseous structures and is especially useful for evaluation of structures within the hoof capsule. This article discusses special ultrasonographic techniques, an overview of MRI image generation, and the use of contrast computed tomography for diagnosis of soft tissue injuries.

COMPUTED TOMOGRAPHIC FINDINGS IN 24 DOGS WITH LIPOSARCOMA.

Computed tomography (CT) continues to become more widely available for assessment of tumors in dogs, yet there are no studies describing the CT appearance of canine liposarcomas. In this retrospective, multicenter study, CT images of dogs with histologically confirmed liposarcomas were reviewed for size, location, attenuation, contrast enhancement, border definition, internal homogeneity, local infiltration, and mineralization. A total of 24 dogs with 26 liposarcomas were sampled. Mean attenuation was +15.2 (SD = 22.3) Hounsfield units (HU) with a range of -36 to +47.5 HU based on representative regions of interest. Twenty tumors (77%) contained focal areas of fat attenuation. All masses enhanced with contrast medium administration, which is distinct from what has been reported previously in infiltrative lipomas. Other CT features associated with canine liposarcomas included heterogeneous internal attenuation (81%) and lack of a clearly defined capsule (38%) suggesting infiltration of local structures. Six tumors (23%) had foci of mineralization. Findings from the current study indicated that liposarcoma should be considered as a differential diagnosis for mixed-attenuation, contrast-enhancing masses in dogs that contain at least one focus of fat attenuation on precontrast images; however, presence of foci of fat attenuation was not a necessary finding for the diagnosis of canine liposarcoma.

Computed tomographic findings in 57 cats with primary pulmonary neoplasia.

Primary pulmonary neoplasia is relatively uncommon in cats and generally has a poor prognosis. In this multicenter, retrospective study of 57 cats with pulmonary neoplasia, the most frequent presenting signs were anorexia/inappetence (39%) and cough (37%). The pulmonary tumors were considered to be incidental findings in 9% cats. In computed tomographic (CT) images, primary pulmonary tumors appeared as a pulmonary mass in 55 (96%) cats and as a disseminated pulmonary lesion without a defined mass in two (4%) cats. Most pulmonary tumors were in the caudal lobes, with 28 (49%) in the right caudal lobe and 17 (30%) in the left caudal lobe. CT features associated with pulmonary tumors included mass in contact with visceral pleura (96%), irregular margins (83%), well-defined borders (79%), bronchial compression (74%), gas-containing cavities (63%), foci of mineral attenuation (56%), and bronchial invasion (19%). The mean (range) maximal dimension of the pulmonary masses was 3.5 cm (1.1-11.5 cm). Additional foci of pulmonary disease compatible with metastasis were observed in 53% cats. Pleural fluid was evident in 30% cats and pulmonary thrombosis in 12% cats. The histologic diagnoses were 47 (82%) adenocarcinomas, six (11%) tumors of bronchial origin, three (5%) adenosquamous cell carcinomas, and one (2%) squamous cell carcinoma. In this series, adenocarcinoma was the predominant tumor type, but shared many features with less common tumor types. No associations were identified between tumor type and CT features. Prevalence of suspected intrapulmonary metastasis was higher than in previous radiographic studies of cats with lung tumors.