Generation and characterization of mesenchymal stem cells from the affected femoral heads of dogs with Legg Calvé Perthes disease.

Background: Canine Legg Calvé Perthes disease (LCPD) occurs during the growth period, and the cause of ischemic necrosis of the femoral head during growth remains unclear. If LCPD-affected femoral head-derived mesenchymal stem cells (LCPD-MSCs) can be generated, they can be used as a new tool for the pathophysiological analysis of canine LCPD. Aim: To generate affected femoral head-derived mesenchymal stem cells (MSCs) from dogs with LCPD and investigate the mRNA expression levels of angiogenesis-related factors and osteogenic differentiation potency of LCPD-MSCs. Methods: This study was performed using affected femoral heads from dogs diagnosed with LCPD and underwent femoral head and neck ostectomy. The necrotic tissue was harvested from the LCPD-affected femoral head and cultured statically (LCPD group, n = 6). Canine bone marrow-derived MSCs (BM-MSCs) were used as controls (control group, n = 6). First, the morphology of the cultured cells was observed, and the expression of CD29, CD34, CD44, CD45, CD90, and major histocompatibility complex class II was analyzed using flow cytometry. Additionally, the trilineage differentiation potency of the LCPD-affected head-derived adherent cells was examined. Furthermore, the expression levels of HIF1A, VEGFA, VEGFB, and PDGFB mRNAs and the bone differentiation potency of LCPD-affected head-derived adherent cells were investigated. Results: LCPD-affected femoral head-derived adherent cells showed a fibroblast-like morphology, and the expression of cell surface antigens was similar to that of BM-MSCs. In addition, LCPD-affected femoral head-derived adherent cells showed the same trilineage differentiation potency as BM-MSCs and were consistent with MSC characteristics. Furthermore, the mRNA expression levels of angiogenesis-related factors could be objectively measured in LCPD-MSCs and those MSCs had bone differentiation potency. Conclusion: In the present study, canine LCPD-MSCs were successfully generated, suggesting their usefulness as a tool for pathological analysis of LCPD in dogs.

A pilot study of microRNA assessment as a means to identify novel biomarkers of spontaneous osteoarthritis in dogs.

MicroRNAs (miRNAs) are important regulators of intercellular signaling and are promising biomarkers in osteoarthritis (OA). In this study, comprehensive analysis was performed to identify miRNAs involved in the pathogenesis of spontaneous OA in dogs. Dogs diagnosed with OA based on radiography and arthroscopy of the stifle joint were included in the OA group. Dogs without any evidence of orthopedic disease were included in the unaffected group. To investigate miRNA expression levels, RNA sequencing analysis (RNA-seq) was performed in synovial tissue (OA group: n = 3, Unaffected group: n = 3) and RT-qPCR was performed in synovial tissue, synovial fluid and serum (OA group: n = 17, Unaffected group: n = 6), and compared between the two groups. The RNA-seq results showed that 57 miRNAs were significantly upregulated and 42 were significantly downregulated in the OA group. Specifically, miR-542 and miR-543 expression levels in the synovial tissue, synovial fluid, and serum were consistently higher in the OA group than in the unaffected group, suggesting that these miRNAs may be used as biomarkers for detecting canine OA. This is the first report to comprehensively analyze the expression patterns of miRNAs in the synovial tissue of dogs with spontaneous OA.

Evaluation of Hindlimb Deformity and Posture in Dogs with Grade 2 Medial Patellar Luxation during Awake Computed Tomography Imaging while Standing.

OBJECTIVE: The aim of this study was to determine the degree of bone deformities and hindlimb postural abnormalities in a standing position in awake Toy poodles with and without grade 2 medial patellar luxation (MPL) using high speed 320-row computed tomography (CT). METHODS: The limbs with grade 2 MPL (MPL-G2 group) and without any orthopaedic disorders (control group) were imaged in a standing position, without sedation or anaesthesia, using CT. In MPL-G2 group, images were obtained when the patella was luxated (G2-L group) and reduced (non-luxation, G2-NL group). Bone morphologies of the femur and tibia were quantified three-dimensionally. Hindlimb standing posture was evaluated by measuring femoral rotation and abduction angles, tibial rotation angle, metatarsal rotation angle, foot rotation angle, angle between the femoral anatomical axis and the mechanical axis of hindlimb and stifle joint line convergence angle. RESULTS: There were no significant differences in bone morphologic parameters between the MPL-G2 group (5 limbs) and the control group (6 limbs). In the G2-NL group, there were no significant hindlimb postural abnormalities. In contrast, in the G2-L group, significant hindlimb postural abnormalities including external rotation of femur, internal rotation of tibia and foot, external rotation of tarsal joint, large stifle joint convergence angle, genu varum and toe-in standing were observed. CONCLUSION: Dogs with grade 2 MPL have no bone deformities but show abnormal standing posture when the patella is luxated.

Functional Anatomy of the Craniolateral and Caudomedial Bundles of the Caudal Cruciate Ligament in Beagle Dogs.

OBJECTIVE: The aim of this study was to investigate functional anatomy of the craniolateral bundle (CrLB) and caudomedial bundle (CdMB) of caudal cruciate ligament in normal Beagles. MATERIALS AND METHODS: Twelve stifle joints of 12 Beagle cadavers that were euthanatized for reasons other than musculoskeletal disease were used. Anatomical characteristics of femoral and tibial attachment regions of the caudal cruciate ligament were described, and location and area of the footprints of the CrLB and CdMB were objectively determined. The CrLB was artificially divided into two equal fibre bundle units and the CdMB was artificially divided into four equal bundle units. Changes in the fibre arrangement were recorded during stifle range of motion, and the tension of each fibre bundle unit at various stifle joint angles was objectively measured. RESULTS: The CrLB was attached to the intercondylar fossa of the femur, and the CdMB was attached to the medial surface of the medial condyle of the femur. The centre of the CrLB attachment site was located craniomedial to that of the CdMB on the tibia. During stifle range of motion, each divided fibre bundle unit in the CrLB and CdMB was twisted externally. The tension of entire CdMB tended to be higher than that of entire CrLB at 180 to 135 degrees of the stifle angles. In full extension angle, the tension of entire CdMB was 1.55 times higher than that of entire CrLB. CLINICAL SIGNIFICANCE: This study contributes to understanding of the biomechanics and the function of the caudal cruciate ligament, which help to elucidate the pathophysiology of caudal cruciate ligament tear and to develop stifle joint stabilization surgery.

Morphological Analysis of Bone Deformities of the Distal Femur in Toy Poodles with Medial Patellar Luxation.

OBJECTIVE: The aim of this study was to evaluate the bone deformities of the distal femur of Toy Poodles with medial patellar luxation (MPL) using computed tomography. METHODS: Computed tomography data from 37 stifles of Toy Poodles (16 normal, 10 grade 2 MPL and 11 grade 4 MPL) were analysed. For trochlear morphology, sulcus angle, medial or lateral trochlear inclination and trochlear angle were compared. For condylar morphology, distance between the apex of the medial or lateral trochlear ridge and the most caudal point of the medial or lateral femoral condyle (MC or LC), the length of the cranial/middle (anterior) part (MC(a) or LC(a')) and caudal (posterior) part (MC(p) or LC(p')) of distal femur and medial or lateral condyle width (MCW or LCW) were measured, and the ratios of medial-to-lateral condyles and anterior-posterior parts were compared. RESULTS: Sulcus angle was significantly higher, and medial trochlear inclination was significantly lower in grade 4 MPL group than other groups. Lateral trochlear inclination was not significantly different among 3 groups. Trochlear angles at proximal trochlear groove were significantly lower in grade 4 MPL group than other groups. MC/LC, MC(a)/LC(a'), MC(a)/MC(p), and LC(a')/LC(p') were significantly lower in grade 4 MPL group than other groups; however, there were no significant differences in MC(p)/LC(p') and MCW/LCW among three groups. CONCLUSION: Hypoplasia of the cranial and middle rather than caudal compartment of the distal femur was the primary morphological abnormality in Toy Poodles with grade 4 MPL.

Treatment of methicillin-resistant Staphylococcus aureus infection following tibial plateau leveling osteotomy in a dog.

BACKGROUND: In recent years, surgical site infections caused by drug-resistant pathogens have emerged as a cause of concern in small animal practice. In this report, methicillin-resistant Staphylococcus aureus (MRSA) infections associated with tibial plateau leveling osteotomy (TPLO) is reported. However, there have been no reports on the treatment of MRSA infection following TPLO in dogs. This case report describes the use of a combination of vancomycin and rifampicin to treat MRSA infection following TPLO in a dog. CASE DESCRIPTION: A 7-year-old spayed female American cocker spaniel was referred for right hind limb lameness that did not improve with conservative treatment. The dog was diagnosed with cranial cruciate ligament rupture, for which TPLO was performed. Once the surgical wound was closed, the dog licked the skin on the surgical site, causing the injury to dehisce. MRSA was detected from the purulent discharge, and chloramphenicol was then administered based on the drug sensitivity test results. Because of the continued drainage, the implants were removed after the bone union of the osteotomy site was observed. Since this did not provide any relief to the existing condition, the antibiotic was changed to vancomycin at 132 days after TPLO surgery, and the infected location was cleaned many times through a drain tube placed into the tibia. However, the infection could not be controlled. Thus, a rifampicin and vancomycin combination was started. As a result, the purulent discharge disappeared and the fistula entirely closed on the 154th day after TPLO surgery. CONCLUSION: A combination of rifampicin and vancomycin may be effective for treating MRSA infection at the surgical site following TPLO surgery that does not heal despite implant removal and administration of vancomycin.

Utility of a novel activity monitor assessing physical activities and sleep quality in cats.

Osteoarthritis in cats is more prevalent because cats are living longer with advancement in veterinary medicine. Objective evaluation of behavioral changes in cats with osteoarthritis can facilitate an early diagnosis. The objective of this study was to investigate the utility of a novel activity monitor for analyzing physical activities and sleep quality in cats. First, a novel activity monitor (Plus Cycle®; JARMeC, Kanagawa, Japan), with a built-in three-directional accelerometer and an air pressure sensor specifically designed for cats, was compared to a human activity monitor (Actical®; Philips Respironics, OR, USA) previously used in cats (n = 10). Second, the validity of the measurement accuracy of the amount of physical activity, the number of vibrations, the number of jumps, and the resting and sleeping time was evaluated using Plus Cycle® in healthy cats (n = 6). Finally, the effects of gender and age of cats and time of day on the amount of physical activity, the number of vibrations, the number of jumps, and the resting and sleeping time were investigated in client-owned cats (n = 61). There were strong correlations between Plus Cycle® and Actical® in total activity (p < 0.05) and activity intensity (p < 0.05). When the physical activities were measured using Plus Cycle® in healthy cats, those data were quantified with high accuracy. In addition, it was also found to be very accurate in discriminating the resting and sleeping conditions of cats. In client-owned cats, there were no significant differences with respect to gender in any measured traits. The amount of physical activity and the number of jumps significantly decreased with the age of the cat. In contrast, the resting and sleeping times significantly increased with the age of the cat. In conclusion, Plus Cycle® can accurately and objectively assess physical activities and sleep quality with age of the cat, suggesting that this novel activity monitor can be used to manage the feline musculoskeletal health.

Functional Anatomy of the Craniomedial and Caudolateral Bundles of the Cranial Cruciate Ligament in Beagle Dogs.

OBJECTIVE: The aim of this study was to investigate detailed functional anatomy of the craniomedial bundle (CrMB) and caudolateral bundle (CdLB) of cranial cruciate ligament (CrCL) for application of intra-articular anatomical reconstruction in dogs. MATERIALS AND METHODS: Twelve stifle joint of 12 Beagle cadavers were used. Anatomical characteristics of femoral attachment regions of the CrCL were determined. Then, the location and area of the footprints of the CrMB and CdLB were objectively measured. Each bundle was equally divided into four fibre bundles, and changes in the fibre arrangement were recorded during stifle range of motion. Additionally, the tension of each fibre bundle at various stifle joint angles was objectively measured. RESULTS: The attachment region of the CrCL clearly differed from that reported in humans. The centre of the CdLB attachment site was located craniodistal to that of the CrMB on the femur and slightly caudal to that of the CrMB on the tibia. The area of the CrMB on the femoral and tibial attachment sites was larger than that of the CdLB. During stifle range of motion, each divided fibre bundle in the CrMB and CdLB was twisted intricately. The tension of the CrMB was higher than that of CdLB. The central part of the CrMB was always tense during stifle range of motion. CLINICAL SIGNIFICANCE: These results may contribute to the application of anatomical reconstruction surgery for CrCL deficient stifle in dogs.

Epidemiologic study of dogs with the displacement or deformity of the medial and lateral fabellae in Japan.

The prevalences of displacement and deformity of the medial and lateral fabellae in dogs were investigated. This was a retrospective epidemiologic study. Radiographs of canine stifle joints (1022 limbs, 534 dogs) were obtained. The images were taken at the Nihon University Animal Medical Center and three private animal hospitals from January 2003 to July 2012. The position and morphology of the medial or lateral fabella were evaluated on the radiographs. The prevalence of displacement of the medial and lateral fabellae was 1.7% and 0.3%, respectively. The prevalence of deformity of the medial and lateral fabellae was 6.9% and 4.6%, respectively. Aplasia or hypoplasia of the fabella was detected more frequently in the medial fabella. On the other hand, a bipartite or multipartite fabella was observed more frequently in the lateral fabella. Nearly all cases of displacement or deformity of the fabella occurred in dogs weighing less than 10 kg. Abnormalities of the fabella were observed in the medial and lateral fabella. We found that abnormal fabellae were closely associated with medial patellar luxation and to a lesser extent with cranial cruciate ligament rupture. No clinical signs were associated with an abnormality of the fabella, with the exception of two dogs with traumatic avulsion of the lateral head of the gastrocnemius muscle.

Evaluation of bone deformities of the femur, tibia, and patella in Toy Poodles with medial patellar luxation using computed tomography.

OBJECTIVES: To evaluate morphological parameters of the femur, tibia, and patella in Toy Poodles with medial patellar luxation (MPL) using three-dimensional (3D) computed tomography (CT) and to compare these parameters between radiography and CT. METHODS: Thirty-five hindlimbs of Toy Poodles were divided into normal and grade 2 and 4 MPL groups. The anatomical and mechanical lateral proximal femoral angle, anatomical and mechanical lateral distal femoral angle (aLDFA, mLDFA), femoral varus angle (FVA), inclination of the femoral head angle, procurvation angle, anteversion angle (AA), frontal angle of the femoral neck, mechanical medial proximal or distal tibial angle, mechanical cranial proximal or distal tibial angle, tibial plateau angle, tibial torsion angle (TTA), Z angle, relative tibial tuberosity width, ratio of the medial distance of tibial tuberosity to the proximal tibial width (MDTT/PTW), patella size, and the patellar ligament length: patellar length (L:P) ratio were evaluated on radiography and 3D CT. RESULTS: The aLDFA, mLDFA, FVA, and TTA were significantly larger and the AA, MDTT/PTW, and patella were significantly smaller in the grade 4 MPL group. There were significant differences in many parameters between imaging tools, and CT was considered less susceptible to potential artefacts and rotational deformities. CLINICAL SIGNIFICANCE: Toy Poodles with grade 4 MPL had significant femoral varus deformity, medial displacement of the tibial tuberosity, internal torsion of the proximal tibia, and hypoplasia of the patella.