原 康

Objective To evaluate the effectiveness of frozen cortical bone allografts (FCBA) in the treatment of severe radial and ulnar atrophic nonunion fractures. Animals Toy breed dogs with nonunion of radial and ulnar fractures (n = 15). Methods Severe atrophic nonunion fractures were treated with FCBA (eight infected and seven non-infected fractures). Radiographs obtained immediately after surgery, and 1, 2, 3, 6 and 12 months later were evaluated and scored for the periosteal reaction at the bone regeneration sites, the healing process in the bone connection areas at both the proximal and distal sites, and the bone remodelling process within the allografts. Results Improvements in the fracture-healing process and weight-bearing function were observed in all cases. Radiographic scores at the bone connection areas and within the allograft improved significantly over time (p &lt 0.05). There were not any significant differences in radiographic scores between the infected and non-infected groups. Clinical Significance Bone reconstruction with FCBA is effective in the treatment of radial and ulnar nonunion fractures associated with large bone defects, regardless of the infection status of the surgical site.

We compared clinical outcomes after ventral fixation in dogs with atlantoaxial instability (AAI) on the basis of the presence or absence of atlantooccipital overlapping (AOO). Of 41 dogs diagnosed with AAI and treated ventral fixation, 12 exhibited AOO (AOO group), whereas 29 did not (non-AOO group). The AOO group had significantly higher neurological scores before (P=0.024) and 1 month after (P=0.033) surgery compared with the non-AOO group however, no significant differences were observed between the groups 2 months after surgery. The presence of complicating AOO affected the clinical signs for dogs with AAI, but did not directly affect the outcome of surgical stabilization of AAI.

Growth plates at each end of vertebral bodies play a pivotal role in longitudinal spinal growth. Epiphyseal closures are formed in adult humans. Although monkeys are frequently employed in bone and disc research, the age of epiphyseal closure has not been well documented. In this study, histological analyses of lumbar vertebral end plates and the surrounding tissue were performed in 11 normal cynomolgus monkeys aged approximately 9 to 15 years, and unclosed growth plate cartilage was detected in all the end plates. The data from this study constitute the first documentation of persistent vertebral growth plate cartilage in cynomolgus monkeys. The persistence of growth plate cartilage in cynomolgus monkeys approximately 15 years of age or younger, which differs from the complete epiphyseal closure exhibited in adult humans, may affect the biomechanical behavior of the spine. This is an important factor to consider in extrapolating the results of spine and intervertebral disc research using cynomolgus monkeys to adult humans.

OBJECTIVE To evaluate and compare morphological characteristics of the dens in atlantoaxial instability (AAI)-predisposed toy-breed dogs (TBDs) with and without AAI and non-AAI-predisposed healthy Beagles. ANIMALS 80 AAI-affected and 40 nonaffected TBDs and 40 Beagles. PROCEDURES Each dog underwent CT examination of the cervical vertebral column. On median 3-D multiplanar reconstruction images, the dens angle (DA) was measured as were the lengths of the dens and the body of the axis; the dens-to-axis length ratio (ratio of the dens length to the axis body length [DALR]) was calculated. Data were compared among dog groups. RESULTS The DALR in nonaffected TBDs and Beagles did not differ significantly. The mean DALR for AAI-affected TBDs was significantly lower than that for nonaffected TBDs. The mean DA of AAI-affected TBDs was significantly greater than that of Beagles and nonaffected TBDs. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that a low DALR might be associated with a high probability of dens abnormalities in TBDs. Additionally, dens length in AAI-affected TBDs appeared to be smaller than that in non-AAI-affected TBDs, given the low DALR in AAI-affected TBDs. Further investigations to determine reference ranges of the DA and DALR and the potential usefulness of those variables as diagnostic markers for AAI in TBDs are warranted.

The purpose of this study was to investigate the expression of bone morphogenetic protein 4 (BMP4) and its receptors, bone morphogenetic protein receptor I (BMPRI) and BMPRII, in the pituitary gland of healthy adult dogs and in those with ACTH-secreting pituitary adenoma. Quantitative polymerase chain reaction analysis showed that the BMP4 messenger RNA expression level in the ACTH-secreting pituitary adenoma samples was significantly lower than that in the normal pituitary gland samples (P = 0.03). However, there were no statistically significant differences between samples with respect to the messenger RNA expression levels of the receptors BMPRIA, BMPRIB, and BMPRII. Double-immunofluorescence analysis of the normal canine pituitary showed that BMP4 was localized in the thyrotroph (51.3 ± 7.3%) and not the corticotroph cells. By contrast, BMPRII was widely expressed in the thyrotroph (19.9 ± 5.2%) and somatotroph cells (94.7 ± 3.6%) but not in the corticotroph cells (P < 0.001, thyrotroph cells vs somatotroph cells). Similarly, in ACTH-secreting pituitary adenoma, BMP4 and BMPRII were not expressed in the corticotroph cells. Moreover, the percentage of BMP4-positive cells was

The purpose of this study was to investigate the expression of bone morphogenetic protein 4 (BMP4) and its receptors, bone morphogenetic protein receptor I (BMPRI) and BMPRII, in the pituitary gland of healthy adult dogs and in those with ACTH-secreting pituitary adenoma. Quantitative polymerase chain reaction analysis showed that the BMP4 messenger RNA expression level in the ACTH-secreting pituitary adenoma samples was significantly lower than that in the normal pituitary gland samples (P = 0.03). However, there were no statistically significant differences between samples with respect to the messenger RNA expression levels of the receptors BMPRIA, BMPRIB, and BMPRII. Doubleimmunofluorescence analysis of the normal canine pituitary showed that BMP4 was localized in the thyrotroph (51.3 +/- 7.3%) and not the corticotroph cells. By contrast, BMPRII was widely expressed in the thyrotroph (19.9 +/- 5.2%) and somatotroph cells (94.7 +/- 3.6%) but not in the corticotroph cells (P < 0.001, thyrotroph cells vs somatotroph cells). Similarly, in ACTH-secreting pituitary adenoma, BMP4 and BMPRII were not expressed in the corticotroph cells. Moreover, the percentage of BMP4-positive cells was also significantly reduced in the thyrotroph cells of the surrounding normal pituitary tissue obtained from the resected ACTH-secreting pituitary adenoma (8.3 +/- 7.9%) compared with that in normal canine pituitary (P < 0.001). BMP4 has been reported to be expressed in corticotroph cells in the human pituitary gland. Therefore, the results of this study reveal a difference in the cellular pattern of BMP4-positive staining in the pituitary gland between humans and dogs and further revealed the pattern of BMPRII-positive staining in the dog pituitary gland. These species-specific differences regarding BMP4 should be considered when using dogs as an animal model for Cushing's disease. (C) 2015 Elsevier Inc. All rights reserved.

<p>Eyes are supplied O₂ through the cornea and vessels of the retina and iris, which are tissues characterized by aerobic metabolism. Meanwhile, there are no reports on the association between iris sphincter contraction and aerobic metabolism. In this paper, we studied the aforementioned association. Eyes from adult pigs of either sex were obtained from a local abattoir. A muscle strip was connected to a transducer to isometrically record the tension. O₂ consumption was measured using a Clark-type polarograph connected to a biological oxygen monitor. Creatine phosphate (PCr) and adenosine triphosphate (ATP) contents were measured in the muscle strips by high-performance liquid chromatography (HPLC). Iris sphincter muscles were measured in resting, contractile or hypoxic phases. Contraction was induced by hyperosmotic 65 mM KCl (H-65K⁺) or carbachol (CCh), and hypoxia was induced by aeration with N₂ instead of O₂ or by addition of sodium cyanide (NaCN). H-65K⁺- and CCh-induced muscle contraction, involved increasing O₂ consumption. Hypoxia and NaCN significantly decreased H-65K⁺- and CCh-induced muscle contraction and/or O₂ consumption and PCr contents. Our results suggest that the contractile behavior in porcine iris sphincter highly depends on mitogen oxidative metabolism.</p>

OBJECTIVE To assess effects of vertebral distraction-fusion techniques at a treated segment (C5-C6) and an adjacent segment (C4-C5) of canine cervical vertebrae. SAMPLE Cervical vertebrae harvested from cadavers of 10 skeletally mature Beagles. PROCEDURES Three models (intact, titanium plate, and polymethylmethacrylate [PMMA]) for stabilization of the caudal region of the cervical vertebrae (C4 through C7) were applied to the C5-C6 vertebral segment sequentially on the same specimens. Biomechanical assessments with flexion-extension, lateral bending, and axial rotational tests were conducted after each procedure. Range of motion (ROM) for a torque load applied with a 6-axis material tester was measured at C4-5 and C5-6 and calculated by use of a 3-D video measurement system. RESULTS In both the plate and PMMA models, ROM significantly increased at C4-5 and significantly decreased at C5-6, compared with results for the intact model. The ROM at C5-6 was significantly lower for the plate model versus the PMMA model in lateral bending and for the PMMA model versus the plate model in axial rotation. Conversely, ROM at C4-5 was significantly higher in axial rotation for the PMMA model versus the plate model. No significant differences were identified in flexion-extension between the PMMA and plate models at either site. CONCLUSIONS AND CLINICAL RELEVANCE Results of this study suggested that vertebral distraction and fusion of canine vertebrae can change the mechanical environment at, and may cause disorders in, the adjacent segment. Additionally, findings suggested that effects on the adjacent segment differed on the basis of the fusion method used.

Obesity is a risk factor for osteoarthritis(OA). To investigate the roles of increased mechanical loading in the onset of obesity-induced OA, knee joints were histologically analyzed after applying a tail suspension (TS) model to a high-fat diet (HFD)-induced OA model. Mice were divided into four groups: normal diet (ND) with normal loading (NL) group; HFD with NL group; ND with TS group; and HFD with TS group. Whole knee joints were evaluated by immunohistological analysis. The infrapatellar fat pad (IPFP) was excised and mRNA expression profiles were compared by qPCR analysis. After twelve weeks of the diet, body weight was increased by HFD in both the NL group and TS group. Upon histological analysis, the irregularity of the surface layer of articular cartilage was observed only in the NL+HFD group. Osteophyte area increased as a result of HFD in both the NL and TS groups, although osteophyte area in the TS+HFD group was smaller than that of the NL+HFD group. In the evaluation of the IPFP by qPCR, adipokines and inflammatory cytokines also increased as a result of HFD. While TGF-beta increased as a result of HFD, the trend was slightly lower in the TS group, in parallel with osteophyte area. To detect apoptosis of articular chondrocytes, TUNEL staining was employed. TUNEL-positive cells were abundantly observed in the articular cartilage in the HFD mice regardless of mechanical loading. IPFP inflammation, enhanced chondrocyte apoptosis, and osteophyte formation were seen even in the TS group as a result of a HFD. In all, these data demonstrate that HFD contributed to osteophyte formation through mechanical loading dependent and independent mechanisms.

Background: Large bone defects in canines usually require assistance to achieve healing. Implantation of osteoinductive factors can promote bone healing, while transplantation of osteoprogenitor cells can enhance bone regeneration. We hypothesized that implantation of an osteoinductive factor, recombinant human bone morphogenetic protein-2 (rhBMP-2), combined with osteoprogenitor cells, bone marrow-derived mesenchymal stromal cells (BMSCs), would synergistically promote bone healing. In this study, we examined the combined effects of Escherichia coli-derived rhBMP-2 and BMSCs on bone healing after implantation into canine ulnar defects. Results: Critical-sized osteoperiosteal segmental defects (2.5 cm) were created in the ulnae of healthy female beagle dogs, and implanted with combinations of E. coli-derived rhBMP-2 (560 or 140 mu g) and autologous BMSCs (10(7), 10(5), or 0 cells). In the present study, 18 forelimbs of nine healthy purpose-bred female beagles were used. All six treatment groups contained three forelimbs, and the animals were euthanized after 12 weeks. The control groups (560 and 140 mu g/0 cells) were cited from our previous study to reduce the number of experimental animals. Radiographically, the regenerated bone width was significantly increased in the 560 or 140 mu g with 10(7) and 10(5) cells groups compared with the 0 cells groups. By quantitative CT, the bone mineral density was higher in the 560 mu g with 10(7) and 10(5) cells groups, while non-uniformity of the bone mineral density was improved in the 560 mu g with 10(7) and 10(5) cells groups and 140 mu g/10(7) cells group. Mechanically, the maximum loads at failure were significantly higher in the 560 mu g with 10(7) and 10(5) cells groups. Histologically, the regenerated bone was well-developed and contained osteocyte-like cells marrow cavities, and vessels. However, the osteoclasts and osteoblasts were hardly observed. The osteocyte-like cell numbers were significantly higher in the 560 mu g with 10(7) and 10(5) cells and 140 mu g with 10(7) and 10(5) cells groups. Conclusions: Implantation of E. coli-derived rhBMP-2 and BMSCs led to significantly enhanced bone formation, with improved bone mineral density and reduced non-uniformity of the regenerated bone. Combined implantation of rhBMP-2 and BMSCs may be useful for promotion of bone healing in critical-sized defects in canines.

Intramedullary bone marrow-derived mononuclear cell (BM-MNC) transplantation has demonstrated neuroprotective effects in the chronic stage of spinal cord injury (SCI). However, no previous study has evaluated its effects in the acute stage, even though cell death occurs mainly within 1 week after injury in all neuronal cells. Moreover, the mechanism underlying these effects remains unclear. We aimed to investigate the survival of intramedullary transplanted allogeneic BM-MNCs and the production of growth factors after transplantation to clarify the therapeutic potential of intramedullary transplanted BM-MNCs and their protective effects in acute SCI. Sprague-Dawley rats were subjected to traumatic SCI and received intramedullary transplantation of EGFP BM-MNCs (n = 6), BM-MNCs (n = 10), or solvent (n = 10) immediately after injury. To evaluate the transplanted BMMNCs and their therapeutic effects, immunohistochemical evaluations were performed at 3 and 7 days post-injury (DPI). BM-MNCs were observed at the injected site at both 3 (683 +/- 83 cells/mm(2)) and 7 DPI (395 +/- 64 cells/mm(2)). The expression of hepatocyte growth factor was observed in approximately 20% transplanted BMMNCs. Some BM-MNCs also expressed monocyte chemotactic protein-1 or vascular endothelial growth factor. The demyelinated area and number of cleaved caspase-3-positive cells were significantly smaller in the BM-MNC-transplanted group at 3 DPI. Hindlimb locomotor function was significantly improved in the BM-MNCtransplanted group at 7 DPI. These results suggest that intramedullary transplantation of BM-MNCs is an efficient method for introducing a large number of growth factor-producing cells that can induce neuroprotective effects in the acute stage of SCI. (C) 2016 Elsevier Ltd. All rights reserved.

Objectives The objectives of this study were to establish a magnetic resonance imaging-based classification system for canine hyperadrenocorticism according to pituitary gland extension, determine indications for trans-sphenoidal hypophysectomy, and clarify the prognosis for each disease grade. Methods A 5-point classification system (Grades 1 to 5) was developed based on tumour extension in dorsal and cranio-caudal directions. Cases were then classified as Type A: no arterial circle of Willis or cavernous sinus involvement and Type B: cases in which these blood vessels were involved. Results Medical records and magnetic resonance imaging data of 37 cases with hyperadrenocorticism were reviewed. Thirty-three cases underwent surgery; 4 Grade 5 cases did not have appropriate indications for surgery, and other therapies were used. Complete resection was achieved for 3, 3, 22 and 1 Grade 1A, 2A, 3A and 3B cases, respectively. Resection was incomplete in 1, 1 and 2 Grade 3A, 3B and 4B cases, respectively. Remission was achieved in 29 cases. Recurrence occurred in 4 cases, all of which were classified as Grade 3. Clinical Significance Dogs with Type A, Grade 1 to 3 hyperadrenocorticism had a good prognosis following trans-sphenoidal hypophysectomy. Grade 3B, 4 and 5 cases may not be suitable for this surgery.

A metabolic syndrome (MetS) is accompanied by hyperuricemia, during which xanthine oxidoreductase (XOR) catalyzes the production of uric acid. In the cohort study, a correlation between uric acid concentration in the synovial fluid and osteoarthritis (OA) incidence is observed. The purpose of our study was to elucidate XOR function in terms of correlation between MetS and OA. Seven week-old male C57BL6J mice were fed normal diet (ND) or high fat diet (HFD) with or without febuxostat (FEB), a XOR inhibitor. HFD stimulated xanthine oxidase activity in the IPFP and the visceral fat. OA changes at the site of the knee joints had progressed due to HFD, but these changes were reduced upon FEB administration. IL-1β expression in the HFD group was increased in accordance with the enhancement of NLRP3 or iNOS expression in the IPFP, whereas it was inhibited by FEB administration. In the organ culture system, when the IPFP was stimulated with insulin, IL-1β expression was increased in accordance with the increase of NLRP3 expression; however, they were reduced by FEB administration. Based on the above results, we showed that inflammasome activation accompanied by an increase in XOR activit

A metabolic syndrome (MetS) is accompanied by hyperuricemia, during which xanthine oxidoreductase (XOR) catalyzes the production of uric acid. In the cohort study, a correlation between uric acid concentration in the synovial fluid and osteoarthritis (OA) incidence is observed. The purpose of our study was to elucidate XOR function in terms of correlation between MetS and OA. Seven week-old male C57BL6J mice were fed normal diet (ND) or high fat diet (HFD) with or without febuxostat (FEB), a XOR inhibitor. HFD stimulated xanthine oxidase activity in the IPFP and the visceral fat. OA changes at the site of the knee joints had progressed due to HFD, but these changes were reduced upon FEB administration. IL-1 beta expression in the HFD group was increased in accordance with the enhancement of NLRP3 or iNOS expression in the IPFP, whereas it was inhibited by FEB administration. In the organ culture system, when the IPFP was stimulated with insulin, IL-1 beta expression was increased in accordance with the increase of NLRP3 expression; however, they were reduced by FEB administration. Based on the above results, we showed that inflammasome activation accompanied by an increase in XOR activity contributed to IPFP inflammation followed by OA progression. (C) 2016 Elsevier Inc. All rights reserved.

OBJECTIVE To histologically evaluate and compare features of myofibers within the elongated soft palate (ESP) of brachycephalic and mesocephalic dogs with those in the soft palate of healthy dogs and to assess whether denervation or muscular dystrophy is associated with soft palate elongation. SAMPLE Soft palate specimens from 24 dogs with ESPs (obtained during surgical intervention) and from 14 healthy Beagles (control group). PROCEDURES All the soft palate specimens underwent histologic examination to assess myofiber atrophy, hypertrophy, hyalinization, and regeneration. The degrees of atrophy and hypertrophy were quantified on the basis of the coefficient of variation and the number of myofibers with hyalinization and regeneration. The specimens also underwent immunohistochemical analysis with anti-neurofilament or anti-dystrophin antibody to confirm the distribution of peripheral nerve branches innervating the palatine myofibers and myofiber dystrophin expression, respectively. RESULTS Myofiber atrophy, hypertrophy, hyalinization, and regeneration were identified in almost all the ESP specimens. Degrees of atrophy and hypertrophy were significantly greater in the ESP specimens, compared with the control specimens. There were fewer palatine peripheral nerve branches in the ESP specimens than in the control specimens. Almost all the myofibers in the ESP and control specimens were dystrophin positive. CONCLUSIONS AND CLINICAL RELEVANCE These results suggested that palatine myopathy in dogs may be caused, at least in part, by denervation of the palatine muscles and not by Duchenne-or Becker-type muscular dystrophy. These soft palate changes may contribute to upper airway collapse and the progression of brachycephalic airway obstructive syndrome.

Objective-To identify characteristics of chondrodystrophoid and nonchondrodystrophoid small-breed dogs with cervical intervertebral disk herniation (IVDH). Design-Retrospective case series. Animals-187 small-breed (&lt;= 15 kg [33 lb]) dogs that underwent surgery because of cervical IVDH. Procedures-Medical records were reviewed for information on breed, sex, age, weight, location of affected intervertebral disks, duration and severity of neurologic signs, and recovery time. Results-55 of the 187 (29.4%) dogs were Beagles. The most frequently affected intervertebral disk was C2-3 (81/253 [32.0%]), and this was the more frequently affected intervertebral disk in dogs of several chondrodystrophoid breeds, including Beagles (29/66 [43.9%]), Dachshunds (13/37 [35.1%]), Shih Tzus (16/41 [39.0%]), and Pekingese (3/10 [30.0%]). However, caudal disks (C5-6 or C6-7) were more frequently affected in Yorkshire Terriers (13/24 [54.2%]) and Chihuahuas (9/13 [69%]). Shih Tzus and Yorkshire Terriers were significantly older at the time of surgery (mean +/- SD age, 9.6 +/- 2.3 years and 9.5 +/- 2.5 years, respectively) than were Pomeranians (6.2 +/- 2.3 years), and Yorkshire Terriers had a significantly higher number of affected disks (2.0 +/- 0.9) than did Dachshunds (1.1 +/- 0.3). Mean recovery time was significantly longer in Yorkshire Terriers (36.7 +/- 13.0 days) than in Beagles (16.5 +/- 17.1 days), Shih Tzus (17.8 +/- 14.5 days), or Chihuahuas (12.2 +/- 7.2 days). Conclusions and Clinical Relevance-Results suggested that there may be breed-specific differences in the characteristics of cervical IVDH in small-breed dogs.

Currently, the most commonly used bioresorbable scaffold is made of beta-tricalcium phosphate (β-TCP); it is hoped that scaffolds made of a mixture of hydroxyapatite (HA) and poly-D/L-lactide (PDLLA) will be able to act as novel bioresorbable scaffolds. The aim of this study was to evaluate the utility of a HA/PDLLA scaffold compared to β-TCP, at a loading site. Dogs underwent surgery to replace a section of tibial bone with a bioresorbable scaffold. After the follow-up period, the scaffold was subjected to histological analysis. The HA/PDLLA scaffold showed similar bone formation and superior cell and tissue infiltration compared to the β-TCP scaffold, as seen after Villanueva Goldner staining. Moreover, silver staining and immunohistochemistry for Von Willebrand factor and cathepsin K demonstrated better cell infiltration in the HA/PDLLA scaffold. The fibrous tissue and cells that had infiltrated into the HA/PDLLA scaffold tested positive for collagen type I and RUNX2, respectively, indicating that the tissue and cells that had infiltrated into the HA/PDLLA scaffold had the potential to differentiate into bone. The HA/PDLLA scaffold is therefore likely to find clinical applica

Osteoarthritis (OA) is a chronic degenerative joint disorder commonly associated with metabolic syndrome. As ageing and obesity has a great impact on the initiation/severity of OA, herein we sought to investigate the involvement of Sirt6 in the crosstalk between ageing and metabolic syndrome/OA. Sirt6 haploinsufficiency in mice promoted the expression of inflammatory cytokines in the IPFP. Enhanced inflammation of the IPFP in the aged Sirt6 +/- HFD group was paralleled with accelerated OA change, including osteophyte growth and chondrocyte hypertrophy. Conversely, mesenchyme-specific Sirt6-deficient mice revealed both attenuated chondrocyte hypertrophy and proteoglycan synthesis, although chondrocyte senescence was enhanced as shown in the aged WT mice. Thus Sirt6 has key roles in the relationship among ageing, metabolic syndrome, and OA. (C) 2015 The Authors. Published by Elsevier Inc.

Degenerative cranial cruciate ligament (CCL) rupture is characterized histologically by degenerating extracellular matrix (ECM) and chondroid metaplasia. Here, we describe the progression of chondroid metaplasia and the changes in the expression of ECM components in canine CCL rupture (CCLR). CCLs from 26 stifle joints with CCLR (CCLR group) and normal CCLs from 12 young beagles (control group) were examined histologically and immunohistochemically for expression of type I (COLD, type II (COLII), type III collagen (COLIII) and Sry-type HMG box 9 (SOX9). Cell density and morphology of CCLs were quantified using hematoxylin eosin staining. The percentage of round cells was higher in the CCLR group than in controls. COLI-positive areas were seen extensively in the connecting fibers, but weakly represented in the cytoplasm of normal CCLs. In the CCLR group, there were fewer COLT-positive areas, but many COLT-positive cells. The percentages of COLII-, COLIII- and SOX9-positive cells were higher in the CCLR group than in controls. The number of spindle cells with perinuclear halo was high in the CCLR group, and most of these cells were SOX9-positive. Deposition of COLT, the main ECM component of ligaments, decreased with increased COLIII expression in degenerated CCL tissue, which shows that the deposition of the ECM is changed in CCLR. On the contrary, expression of SOX9 increased, which may contribute to the synthesis of cartilage matrix. The expression of COLII and SOX9 in ligamentocytes showed that these cells tend to differentiate into chondrocytes.

Objectives: In dogs with deep analgesia caused by acute spinal cord injury from thoracolumbar disk herniation, autologous bone marrow mononuclear cell transplant may improve recovery. The purpose of the present study was to evaluate autologous bone marrow mononuclear cell transplant in a dog that had paraplegia and deep analgesia caused by chronic spinal cord injury. Materials and Methods: Autologous bone marrow mononuclear cell transplant was performed in a dog having paraplegia and analgesia for 3 years that was caused by a chronic spinal cord injury secondary to Hansen type I thoracolumbar disk herniation. Functional recovery was evaluated with electrophysiologic studies and the Texas Spinal Cord Injury Scale. Results: Somatosensory evoked potentials were absent before transplant but were detected after transplant. Functional improvement was noted (Texas Spinal Cord Injury Scale: before transplant, 0; after transplant, 6). No adverse events were observed. Conclusions: Autologous bone marrow mononuclear cell transplant into the subarachnoid space may be a safe and beneficial treatment for chronic spinal cord injury in dogs.

Objective: To create a canine model of excessive tibial plateau angle (eTPA) and assess the chondroid metaplasia and extracellular matrix alteration in the cranial cruciate ligament. Methods: Seven mature female Beagles were included. Cylindrical osteotomy was performed bilaterally in the proximal tibia. The TPA was increased to approximately 40 degrees in the left tibia (eTPA stifle) and left unchanged in the right tibia (control stifle). Exercise stress was started at three months postoperatively, and at 12 months postoperatively the dogs were euthanatized and the cranial cruciate ligaments were collected. The specimens were subjected to haematoxylin and eosin staining to assess the ligamentocyte morphology and immunostaining to assess the type I (COLI), type II (COLII), and type III (COLIII) collagen, and the sry-type HMG box 9 (SOX9) staining. Results: Macroscopic cranial cruciate ligament injury was absent in six dogs but present in the eTPA stifle of one dog, which was excluded from the analysis. The ligamentocyte density decreased and the percentage of round ligamentocytes increased in the eTPA stifles. The COLII, COLIII, and SOX9 staining increased significantly and COLI deposition decreased in the eTPA stifles compared to the control stifle. Clinical significance: The extracellular matrix changed, COLI deposition decreased, and COLIII and SOX9 staining increased in the cranial cruciate ligament of the eTPA stifles. SOX9 may contribute to COLII synthesis in the extracellular matrix of the cranial cruciate ligament in eTPA stifles, and eTPA may promote chondroid metaplasia and extracellular matrix alteration.

Intervertebral disc degeneration (IVDD) greatly affects the quality of life. The nucleus pulposus (NP) of chondrodystrophic dog breeds (CDBs) is similar to the human NP because the cells disappear with age and are replaced by fibrochondrocyte-like cells. Because IVDD develops as early as within the first year of life, we used canines as a model to investigate the in vitro mechanisms underlying IVDD. The mechanism underlying age-related IVDD, however, is poorly understood. Several research groups have suggested that Wnt/β-catenin signaling plays an important role in IVDD. However, the role of Wnt/β-catenin signals in IVD cells is not yet well understood. Here, we demonstrate that Wnt/β-catenin signaling could enhance Runx2 expression in IVDD and lead to IVD calcification. Nucleus pulposus (NP) tissue was obtained from Beagle dogs after evaluation of the degeneration based on magnetic resonance imaging (MRI). Histological analysis showed that lack of Safranin-O staining, calcified area, and matrix metalloproteinase (MMP) 13-positive cells increased with progression of the degeneration. Furthermore, the levels of β-catenin- and Runx2-positive cells also increased. Real-time revers

Currently, the most commonly used bioresorbable scaffold is made of beta-tricalcium phosphate (beta-TCP); it is hoped that scaffolds made of a mixture of hydroxyapatite (HA) and poly-D/L-lactide (PDLLA) will be able to act as novel bioresorbable scaffolds. The aim of this study was to evaluate the utility of a HA/PDLLA scaffold compared to beta-TCP, at a loading site. Dogs underwent surgery to replace a section of tibial bone with a bioresorbable scaffold. After the follow-up period, the scaffold was subjected to histological analysis. The HA/PDLLA scaffold showed similar bone formation and superior cell and tissue infiltration compared to the beta-TCP scaffold, as seen after Villanueva Goldner staining. Moreover, silver staining and immunohistochemistry for Von Willebrand factor and cathepsin K demonstrated better cell infiltration in the HA/PDLLA scaffold. The fibrous tissue and cells that had infiltrated into the HA/PDLLA scaffold tested positive for collagen type I and RUNX2, respectively, indicating that the tissue and cells that had infiltrated into the HA/PDLLA scaffold had the potential to differentiate into bone. The HA/PDLLA scaffold is therefore likely to find clinical application as a new bioresorbable scaffold.

Conjunctival epithelial and goblet cell P2Y2 nucleotide receptors regulate ion transport and secretory function. Diquafosol is a P2Y2 purinergic receptor agonist that stimulates secretion of aqueous tear components from conjunctival epithelial cells and secretion of mucin from conjunctival goblet cells. In humans suffering from keratoconjunctivitis sicca (dry eye), topical administration of diquafosol improves corneal epithelial integrity and stabilises the tear film. The aim of the present study was to investigate P2Y2 receptor expression and to determine the effect of topical administration of diquafosol on mucin and aqueous tear production in dogs. Canine conjunctival P2Y2 receptor expression was evaluated by Western blotting and immunohistochemical analysis. The effect of diquafosol on mucin secretion was evaluated by examining mucin-5 subtype AC (MUC5AC) concentration in tears. The effect of diquafosol on aqueous secretions was evaluated by performing the Schirmer tear test (STT) and phenol red thread test. Expression of the P2Y2 receptor was confirmed in canine bulbar and palpebral conjunctivae and receptors were identified at the conjunctival epithelial and goblet cell surface. Tear MUC5AC concentration significantly increased after administration of 3% diquafosol ophthalmic solution, although neither STT nor phenol red thread test values showed any significant change after diquafosol instillation. Topical ocular administration of 3% diquafosol might improve corneal epithelial disorders in dogs through stabilisation of the tear film, by virtue of an increase in MUC5AC secretion.

Objectives To compare data for French Bulldogs and Dachshunds that had hemilaminectomy for thoracolumbar intervertebral disc extrusion (T-L IVDE) by 1 surgeon and to evaluate the association between IVDE and congenital vertebral anomalies. Design Retrospective case series. Animals French Bulldogs (n = 47) and 671 Dachshunds. Methods Age, gender, vertebral anomaly, kyphosis/kyphoscoliosis, IVDE site, non-recovery and progressive hemorrhagic myelomalacia development from grade 5 (paraplegia without deep nociception) were compared between the 2 breeds. Results French Bulldogs were significantly younger (P = .00001), more likely to be male (P = .023), and more likely to have a congenital vertebral anomaly and kyphosis/kyphoscoliosis (P &lt; .00001) than Dachshunds. The frequencies of French Bulldogs with IVDE within typical sites (T11-L3) were significantly lower (P = .0005) and within caudal sites (L3-L7) significantly higher (P = .0001) compared with Dachshunds. None of the French Bulldogs had IVDE within the kyphotic/kyphoscoliotic segment. The frequency of lumbar IVDE (L1-L5) in French Bulldogs with kyphosis/kyphoscoliosis was significantly higher (P = .003) compared with French Bulldogs without kyphosis/kyphoscoliosis. In grade 5 dogs, the risk of developing progressive hemorrhagic myelomalacia in French Bulldogs was significantly higher (P = .03) than in Dachshunds. Conclusion The distribution of IVDE site in French Bulldogs within the thoracolumbar and lumbar spine was different from Dachshunds. IVDE sites were not located at the sites of vertebral anomaly. French Bulldogs appeared to have T-L IVDE at younger ages, with higher male predisposition and higher risk of developing progressive hemorrhagic myelomalacia from grade 5 compared with Dachshunds.

Hydroxyapatite (HA)/poly-l-lactide(PLLA) composite biomaterials are available for orthopedic applications, but bioresorption and cell-mediated inflammation in bone cortex are unknown. We conducted an 84-month follow-up study with Beagle dogs that were subjected to implants with either PLLA (left femur) or HA/PLLA (right femur). Histological and radiographic analysis showed that HA/PLLA screws induced significant increases in HA content from 36 months onward and complete burr hole closure at 60 months, whereas PLLA screws did not. Moreover, PLLA screws induced more severe fibrous tissue and histiocyte infiltration. HA/PLLA screws promote earlier burr hole replacement and have superior biocompatibility compared to PLLA screws.

近年、動物医療においてもリハビリテーションの重要性が認識され、治療行為のひとつとして注目を集めつつある。本研究では前十字靭帯断裂に馬尾症候群を併発した一事例を対象に術後リハビリテーションを行い、その有用性について検討した。術後早期からアイシングや他動的関節可動域運動(PROM運動)、マッサージ療法、温熱療法、水治療法を実施したところ、対照群と比べて膝関節の関節可動域の減少が小さく、筋肉量の減少は見られなかった。また、フォースプレートで解析した歩様回復までの期間は、対照群の3ヶ月に対し2ヶ月と短かった。これらの結果より、TPLO実施後早期からのリハビリテーションの効果は高く、術後回復の促進に有用であることが示唆された。日本の獣医療においてリハビリテーションがより広く普及し、麻痺や痛みを抱える患者動物とそのオーナーの苦しみを、少しでも減らす手助けとなることを期待する。