[show abstract][hide abstract] ABSTRACT: Aim of the study was to verify the clinical and morphological effects of intra-articular stanozolol or placebo treatment, lasting 3 and 9months, in sheep in which a femoro-tibial osteo-arthritis (OA) were surgically induced (medial bilateral meniscectomy). Twenty healthy sheep divided into four groups and two control animals group, after surgical medial bilateral meniscectomy, were weekly injected in femoral-tibial joint (FTJ) with stanozolol or placebo. Lameness evaluation was performed and synovial fluid was collected from all sheep at each treatment time. Necropsies were performed after 3 or 9month as described in experimental design. Gross pathologies were described and specimen tissues collected from femoro-tibial articular joints were processed for routine histological examination. The gross anatomy of the FTJ was well-preserved in stanozolol-treated sheep; this also applied to the histological features of articular cartilage. Joint aseptic inflammation and fibrosis were observed in placebo-treated sheep, associated with a different degree of severity of condylar and tibial plate cartilage degeneration. Stanozolol intra-articular treatment reduces osteophytes formation and subchondral bone reaction and promotes articular cartilage regeneration.
Research in Veterinary Science 01/2013; · 1.77 Impact Factor
[show abstract][hide abstract] ABSTRACT: Purpose. This study evaluated the efficacy of a regenerative approach using mesenchymal stem cells (MSCs) and CAD-CAM customized pure and porous hydroxyapatite (HA) scaffolds to replace the temporomandibular joint (TMJ) condyle. Methods. Pure HA scaffolds with a 70% total porosity volume were prototyped using CAD-CAM technology to replace the two temporomandibular condyles (left and right) of the same animal. MSCs were derived from the aspirated iliac crest bone marrow, and platelets were obtained from the venous blood of the sheep. Custom-made surgical guides were created by direct metal laser sintering and were used to export the virtual planning of the bone cut lines into the surgical environment. Sheep were sacrificed 4 months postoperatively. The HA scaffolds were explanted, histological specimens were prepared, and histomorphometric analysis was performed. Results. Analysis of the porosity reduction for apposition of newly formed bone showed a statistically significant difference in bone formation between condyles loaded with MSC and condyles without (P < 0.05). The bone ingrowth (BI) relative values of split-mouth comparison (right versus left side) showed a significant difference between condyles with and without MSCs (P < 0.05). Analysis of the test and control sides in the same animal using a split-mouth study design was performed; the condyle with MSCs showed greater bone formation. Conclusion. The split-mouth design confirmed an increment of bone regeneration into the HA scaffold of up to 797% upon application of MSCs.
BioMed research international. 01/2013; 2013:549762.
[show abstract][hide abstract] ABSTRACT: In this study, rapid CAD-CAM prototyping of pure hydroxyapatite to replace temporomandibular joint condyles was tested in sheep. Three adult animals were implanted with CAD-CAM-designed porous hydroxyapatite scaffolds as condyle substitutes. The desired scaffold shape was achieved by subtractive automated milling machining (block reduction). Custom-made surgical guides were created by direct metal laser sintering and were used to export the virtual planning of the bone cut lines into the surgical environment. Using the same technique, fixation plates were created and applied to the scaffold pre-operatively to firmly secure the condyles to the bone and to assure primary stability of the hydroxyapatite scaffolds during masticatory function. Four months post-surgery, the sheep were sacrificed. The hydroxyapatite scaffolds were explanted, and histological specimens were prepared. Different histological tissues penetrating the scaffold macropores, the sequence of bone remodeling, new apposition of bone and/or cartilage as a consequence of the different functional anatomic role, and osseointegration at the interface between the scaffold and bone were documented. This animal model was found to be appropriate for testing CAD-CAM customization and the biomechanical properties of porous, pure hydroxyapatite scaffolds used as joint prostheses.
Journal of Biomaterials Applications 04/2012; · 2.64 Impact Factor
[show abstract][hide abstract] ABSTRACT: No endoscopic examination of the nasolacrimal duct has been described before. In contrast with other imaging techniques, endoscopy provides a direct inspection of the intralumen and ductal mucosa in standing sedation.
To provide a reference against which the endoscopic and clinical features of obstructive nasolacrimal disease in the horse may be compared.
Endoscopic examination of the nasolacrimal duct was performed in 10 French Thoroughbred bay mares with a 3 mm shaft diameter flexible fibrescope. The duct was divided into 3 zones (1, 2 and 3) from the nostril to the lacrimal sac.
Endoscopic examination of the nasolacrimal duct appeared to be quite easy to perform in Zones 1 and 2, while the ease or difficulty of examining Zone 3 depended on the weight and size of the horse for the position of the endoscope in the lacrimal canal.
Endoscopic inspection has provided the possibility of a novel diagnostic approach to conditions of the nasolacrimal duct in horses.
Endoscopic examination of the nasolacrimal duct could help to diagnose and treat nasolacrimal diseases by performing sample withdrawal and duct irrigation.
[show abstract][hide abstract] ABSTRACT: Gastrointestinal stromal tumours (GISTs) represent a distinctive group of primary mesenchymal tumours of the gastrointestinal tract identified immunohistochemically by expression of CD117. A 10-year-old neutered female domestic shorthair cat with a history of recurrent vomiting was examined. The presence of a gastric mass was recognized and a laparotomy was performed. Cytological examination was consistent with a low-grade malignant mesenchymal tumour and histopathological investigation suggested myogenic differentiation of tumour cells. The diagnosis of GIST was confirmed by immunohistochemical expression of CD117. Sequence analysis of the KIT gene identified a deletion in exon 11. The same mutation is found often in human GISTs.
Journal of comparative pathology 02/2011; 145(2-3):152-7. · 1.73 Impact Factor
[show abstract][hide abstract] ABSTRACT: Minimum criteria for the characterisation of human mesenchymal stem cells (MSC) are a) adhesion to the plastic when maintained under culture conditions; b) expression of CD105, CD73, and CD90, and no expression for CD45, CD34, and CD14; and c) differentiation into osteoblasts, adipocytes, and chondroblasts in vitro. One major difficulty in characterising equine MSC is the absence of specific monoclonal antibodies and evidence that certain markers from other species do not cross-react with the equine species. The aim of this work was to isolate, cultivate, differentiate, and conduct cellular characterisation of MSC derived from equine adipose tissue (AT) and bone marrow (BM). Adipose tissue collection was performed at the base of the horses' tails, and BM was aspirated from the iliac crest. Mononuclear cell fraction was isolated and cultured as previously described by (Colleoni et al. 2009 Vet. Res. Commun. 33, 811-821). Chondrogenic, osteogenic, and adipogenic differentiation were performed in monolayer culture, and evidence for differentiation was made by morphological and cytological evaluations. For molecular characterisation, cells were treated with trypsin, washed with PBS, and fixed with Reagent 1 (Intraprep Kit, Beckman Coulter, Miami, FL, USA), following the manufacturer's instructions. Samples, after washings, were incubated for 20min at room temperature with CD105, CD90, CD44, CD45, CD34, CD14, and CD73 mAbs, directly conjugated to fluorescein isothiocyanate, PE, or APC (Beckman Coulter). Appropriate conjugate isotype controls were applied (Beckman Coulter). After staining, cells were washed twice with PBS, and fluorescence intensity was evaluated with a FC500 two-laser equipped cytometer (Beckman Coulter). Results were further analysed with the CXP dedicated program. Samples volumes were 68±23.6mL for BM and 5.6±1.1g for AT; in both AT and BM, the isolation rate was 100% (AT: 4/4; BM: 5/5). Undifferentiated cells were passaged up to 8 times for AT and 5 times for BM; population-doubling times (DT) were calculated, and data were analysed by ANOVA (Statistica for Windows, Stat Soft Inc., Tulsa, OK, USA). No significant differences (P>0.05) were found between DT of all passages. The DT was greater (P<0.05) for BM (3.2±1.5) than for AT (1.3±0.7). By passage 8, AT MSC underwent 37.3±4.6 cell-doublings (CD); by passage 5, BM MSC underwent 26.2±5.03 CD. Positive von Kossa and Alizarin Red staining confirmed osteogenesis. Alcian blue staining illustrated chondrogenesis, and positive Oil Red O staining suggested adipogenesis. The AT and BM MSC were positive for CD90, CD44, and CD105; all cell lines were negative for haematopoietic markers such as CD34, CD14, and CD45. Although marker CD73 expresses reaction in other studies involving MSC in different species, it did not cross-react with equine AT and BM MSC. Results obtained revealed the immunophenotypic characterisation of the surface of isolated and cultivated MSC, classifying these cells as a promising type of progenitor cells that can be applied in equine cellular therapy.
Reproduction Fertility and Development 01/2011; 23(1):250-251. · 2.58 Impact Factor
[show abstract][hide abstract] ABSTRACT: Centronuclear myopathy (CNM) is an autosomal recessive hereditary disease affecting Labrador Retriever dogs. The disease is characterized by muscle lesions, typically encompassing reduction in the number and atrophy of type II fibers, and is caused by a short interspersed repeat element insertion in exon 2 of the protein tyrosine phosphatase-like member A. The actual allele frequency is unknown; a study was undertaken to ascertain it using a convenience-sample population composed of 217 Labrador Retrievers. In addition to 3 subjects already diagnosed with CNM, used as positive controls for polymerase chain reaction, only 2 unrelated dogs were heterozygous wild-type/mutation (wild-type/mut). Thus, the frequency of the CNM allele observed in the present study was 1.8% and 0.47% when including and excluding the 3 mut/mut homozygous cases, respectively. Based on the Hardy-Weinberg exact test (P = 1.00), the genotype frequency without the CNM-affected dogs was in agreement with the Hardy-Weinberg equilibrium. Assuming the Hardy-Weinberg equilibrium law, the expected frequency of the homozygous mutated genotype was calculated to be approximately 0.00005, which corresponds to 1 case of CNM out of 20,000 dogs. In conclusion, the present study indicates that the CNM allele is present but rare in a convenience sample of Labrador Retrievers in Italy.
Journal of veterinary diagnostic investigation: official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 01/2011; 23(1):124-6. · 1.18 Impact Factor
[show abstract][hide abstract] ABSTRACT: The distribution and chemical phenotypes of sympathetic and dorsal root ganglion (DRG) neurons innervating the equine ileocecal junction (ICJ) were studied by combining retrograde tracing and immunohistochemistry. Immunoreactivity (IR) for tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DBH), neuronal nitric oxide synthase (nNOS), calcitonin gene-related peptide (CGRP), substance P (SP), and neuropeptide Y (NPY) was investigated. Sympathetic neurons projecting to the ICJ were distributed within the celiac (CG), cranial mesenteric (CranMG), and caudal mesenteric (CaudMG) ganglia, as well as in the last ganglia of the thoracic sympathetic chain and in the splanchnic ganglia. In the CG and CranMG 91 +/- 8% and 93 +/- 12% of the neurons innervating the ICJ expressed TH- and DBH-IR, respectively. In the CaudMG 90 +/- 15% and 94 +/- 5% of ICJ innervating neurons were TH- and DBH-IR, respectively. Sympathetic (TH-IR) fibers innervated the myenteric and submucosal ganglia, ileal blood vessels, and the muscle layers. They were more concentrated at the ICJ level and were also seen encircling myenteric plexus (MP) and submucosal plexus (SMP) descending neurons that were retrogradely labeled from the ICJ. Among the few retrogradely labeled DRG neurons, nNOS-, CGRP-, and SP-IR nerve cells were observed. Dense networks of CGRP-, nNOS-, and SP-IR varicosities were seen around retrogradely labeled prevertebral ganglia neurons. The CGRP-IR fibers are probably the endings of neurons projecting from the intestine to the prevertebral ganglia. These findings indicate that this crucial region of the intestinal tract is strongly influenced by the sympathetic system and that sensory information of visceral origin influences the sympathetic control of the ICJ.
The Journal of Comparative Neurology 10/2010; 518(19):4046-66. · 3.66 Impact Factor
[show abstract][hide abstract] ABSTRACT: Nasal diseases are very common in dogs and rhinoscopy is often required for a definitive diagnosis. Rhinoscopy, while superficial in nature, can guide the clinician to the final diagnosis. In this study, rhinoscopy was performed on 54 dogs with symptoms of chronic nasopharyngeal disease. The endoscopic diagnosis of neoplasia or chronic nasal inflammation was validated with histological examination of pathological samples, in order to evaluate the degree of concordance between endoscopic findings and histological diagnosis. The agreement between endoscopy and histology was tested by application of Cohen's kappa coefficient. We conclude that correlation between endoscopic results and histological diagnosis, expressed by a Cohen's kappa coefficient of 0.73, is only possible with a constant cooperation between the clinician and the pathologist.
[show abstract][hide abstract] ABSTRACT: In recent years, gastric ulceration has been recognized as a common, possibly performance-limiting disease, of adult horses. The aim of this study was to compare endoscopic features, histological diagnosis, and mRNA levels of various cytokines (TNF-alpha, IL-1beta, and IL-13) from horse gastric biopsies. Eleven horses suffering from equine gastric ulcer syndrome and seven horses with normal histological gastric features were assessed. No correlation between endoscopic features and histology (i.e., the gold standard) was observed. Based on histological diagnosis, a significant (p < 0.05) increase in cytokine mRNA levels (specifically, TNF-alpha and IL-13) was observed in horses affected by equine ulcerative gastric syndrome.
[show abstract][hide abstract] ABSTRACT: This case report documents uroperitoneum due to bladder and pelvic urethral rupture in a 9 h old Standardbred colt born after severe dystocia. Three surgical procedures were required to manage rupture of the urinary bladder, urethral rupture, and ventral abdominal hernia with evisceration of a loop of jejunum. After 40 days of hospitalisation the colt was discharged without any further complication.
[show abstract][hide abstract] ABSTRACT: Ocular ultrasonography in horses represents a valuable imaging diagnostic tool for the diagnosis of intraocular and periocular diseases, particularly when cornea or lens opacities preclude ophthalmoscopy of deeper structures. The authors studied normal and pathological aspects of the eye. Nineteen horses with opacities of the dioptric structures underwent an ultrasound examination. This technique allowed the diagnosis of a posterior synechia (1), cataracts (8), vitreous opacities (6), retinal detachment (3), and a foreign body (1). Ultrasonography provided helpful information about the structure and vascular pattern when the conventional ophthalmic evaluation was unable to achieve a correct diagnosis.
Journal of Equine Veterinary Science - J EQUINE VET SCI. 01/2010; 30(3):150-154.
[show abstract][hide abstract] ABSTRACT: In horses, morpho-functional studies related to the enteric nervous system (ENS) controlling the sphincters are lacking.
To investigate immunohistochemically the morphology, distribution, density, phenotypes and projections of neurons controlling the ileocaecal junction (ICJ).
Two young horses were anaesthetised and underwent midline laparotomy. The neuronal retrograde fluorescent tracer Fast Blue (FB) was injected into the wall of the ICJ. A post surgical survival time of 30 days was used. Following euthanasia, the ileum and a small portion of caecum were removed. Cryosections were used to investigate the immunoreactivity (IR) of the neurons innervating the ICJ for choline acetyltransferase (ChAT), neuronal nitric oxide synthase (nNOS), substance P (SP), calcitonin gene-related peptide (CGRP) and neurofilament NF200kDa (NF).
Ileal FB-labelled neurons innervating the ICJ were located in the myenteric plexus (MP) and submucosal plexus (SMP) up to 48 cm and 28 cm, respectively, from the point of the FB injections. Descending MP and SMP neurons were nitrergic (54 +/- 11% and 68 +/- 4%, respectively), cholinergic (60 +/- 19% and 82 +/- 11%, respectively), NF-IR (54 +/- 9% and 78 +/- 21%, respectively), and SP-IR (about 20% in both the plexuses). CGRP-IR was expressed only by SMP descending neurons (45 +/- 21%). In both the plexuses descending neurons coexpressing nNOS- and ChAT-IR were also observed (25 +/- 11% and 61 +/- 27%, respectively).
The presence of ileal long projecting neurons innervating the ICJ suggests that they are critical for its modulation. Consequently, in bowel diseases in which the resection of the terminal jejunum and proximal ileum are required, it is preferable, whenever possible, to conserve the major portion of the ileum.
The knowledge of the phenotype of ENS neurons of the ileum might be helpful for developing pharmaceutical treatment of the ICJ motility disorders.
[show abstract][hide abstract] ABSTRACT: Mesenchymal stem cells (MSC) are increasingly used as therapeutical aid for the orthopaedic injuries in the horse. MSC populate different tissues but the most commonly used for clinical purposes are isolated from bone marrow or adipose tissue. The first objective of this study was to investigate if the donor animal, the tissue of origin and the technique of isolation could influence the number of MSC available for transplantation after a short-term expansion. The second aim was to devise a culture system capable of increasing MSC lifespan and we tested the effect of basic fibroblast growth factor (bFGF). Results indicate that MSC can be efficiently isolated from both sources and supplementation of bFGF enhances proliferation rate maintaining differentiation potential. In addition, this study shows that collection, expansion and storage of frozen MSC can be performed for later therapeutic use.
Veterinary Research Communications 06/2009; 33(8):811-21. · 1.08 Impact Factor
[show abstract][hide abstract] ABSTRACT: Thymolipomas are unusual benign neoplasms composed of mature adipose and thymic tissue that represent less than 10% of all thymic tumours in man. A thymofibrolipoma is a histological variant of thymolipoma in which there is an additional abundance of fibrous connective tissue. Thymolipoma has been reported previously in one dog and one cat, but thymofibrolipoma has never been described. This report documents the clinical and pathological features of two cases of canine thymofibrolipoma.
Journal of comparative pathology 06/2009; 141(1):74-7. · 1.73 Impact Factor
[show abstract][hide abstract] ABSTRACT: This paper describes the morphology and distribution of the enteric nervous system (ENS) cells and fibres immunoreactive for choline acetyltransferase (ChAT), neuronal nitric oxide synthase (nNOS), substance P (SP), calcitonin gene-related peptide (CGRP), NF200kDa (NF200), and S100 protein. The percentages of subclasses of enteric neurons in the total neuronal population were investigated by the use of anti-PGP 9.5 or anti-NSE antibodies. ChAT-IR myenteric plexus (MP) and submucosal plexus (SMP) neurons were 66+/-7% and 74+/-15%, respectively, whereas those cells expressing nNOS-IR were 38+/-7% and 5+/-1%, respectively. MP and SMP neurons expressing both phenotypes were also present. SP-IR was expressed by 14+/-13% of MP and 66+/-8% of SMP neurons whereas CGRP-IR was observed only in the SMP (43+/-6%). NF200-IR was expressed by 61+/-15% and 91+/-10% of the MP and SMP neurons, respectively. The majority of the CGRP-IR SMP neurons expressed also SP-IR. Almost all SP-IR neurons in both the plexuses were cholinergic. The present study quantifies the main neuronal subpopulations of the ENS of the horse ileum; these data might be utilized to understand the neuronal modifications which occur in several gastrointestinal tract disorders.
Research in Veterinary Science 05/2009; 87(2):177-85. · 1.77 Impact Factor
[show abstract][hide abstract] ABSTRACT: Rupture of the lateral lobe of the proximal tendon of the biceps brachii muscle was diagnosed in an Arabian horse. To the authors' knowledge, this is the only report of this condition in horses. Although clinical signs were helpful in the identification of the location of disease, ultrasonographic examination was a more definitive and non- invasive means of diagnosing the pathological condition. Bursoscopic examination of the intertubercular bursa was also useful in obtaining confirmation of the diagnosis, and for debridement and lavage of the bursa.
Veterinary and Comparative Orthopaedics and Traumatology 02/2009; 22(3):253-5. · 1.01 Impact Factor
[show abstract][hide abstract] ABSTRACT: To report sartorius muscle contracture in a dog.
A 5-year-old, male, German Shepherd dog.
After a 2-month history of right hindlimb lameness was treated by tenectomy of the caudal sartorius muscle for sartorius muscle contracture.
One month after surgery there was complete remission of clinical signs; however, after 12 months, there was partial reunion of sartorius caudal belly and onset of gracilis muscle contracture.
Contracture of the sartorius muscle in dogs is seemingly rare and similar to gracilis muscle contracture with which it may be confused. Differentiation requires knowledge of muscular anatomy, careful palpation, and surgical inspection.
Sartorius muscle contracture can be treated by tenectomy or removal of contracted muscle tissue, however, long-term outcome suggests the need to develop approaches that prevent recurrence caused by scarring of transected tissues.
Veterinary Surgery 03/2008; 37(2):149-52. · 1.24 Impact Factor