Comparison of Imaging Methods and Population Pattern in Dogs with Spinal Diseases in Three Periods between 2005 and 2022: A Retrospective Study.

The aim of this study was the long-term comparison of the imaging methods used in dogs with neurologic diseases related to the spine and spinal cord. We also compared the occurrence of neurological diseases according to the localization, gender, age, and breed. As the availability of magnetic resonance imaging (MRI) has increased over the years, resulting in increased diagnostic and therapeutic success rates, the study was divided into three time periods (2005-2014, 2015-2018, and 2019-2022). Our results suggest changes in the population structure of the dogs studied and changes in the use of diagnostic methods that directly or indirectly influence the choice and success rate of therapy. Our results may be of interest to owners, breeders, practicing veterinarians, and insurance companies.

Inter- and Intra-Observer Variations in Radiographic Evaluation of Pelvic Limbs in Yorkshire Terriers with Cranial Cruciate Ligament Rupture and Patellar Luxation.

The main aims of the study were to describe bone alignment differences in Yorkshire Terriers (YT) with cranial cruciate ligament rupture (CCLR), patellar luxation (PL), or with a combination of both (CCLR + PL); to verify the theory of increased strain on cranial cruciate ligament (CrCL) due to PL as a predisposing factor; and to evaluate intra- and inter-observer variability of the protocols developed for measurement of femoral and tibial alignment in Yorkshire Terriers. Fifty-five hindlimbs of YT were divided into four groups: Control, CCLR, PL, and CCLR + PL. Thirty parameters were radiographically evaluated including hip joint, femoral, tibial, and intercondylar fossa (ICF) parameters. Three observers evaluated all parameters on two separate occasions with a twelve-week interval between measurements. Significant differences in conjunction with CCLR and PL theories between groups were noticed in: Norberg angle (NA), quadriceps angle (Qa), anatomic lateral distal femoral angle (aLDFA), femoral varus (FVA), mechanical cranial proximal tibial angle (mCrPTA), mechanical caudal proximal tibial angle (mCdPTA), tibial plateau angle (TPA), distal tibial axis/proximal tibial axis angle (DPA). Some interesting findings are the similarity of values between Control and CCLR vs. CCLR + PL and PL groups in Na, Qa, aLDFA; between CCLR + PL and PL in FVA and a significantly lower age of dogs in CCLR + PL compared to CCLR group. Based on our results, we can conclude that YT with different clinical findings have differing bone morphology. Moreover, these findings may support PL as a predisposing factor for CCLR in small breeds. Measurements in which excellent inter- observer agreement was achieved may be used for surgical planning or for further discussions.

Relationship of mRNA Expression of Selected Genes in Peripheral Blood and Synovial Fluid in Cranial Cruciate Ligament Deficient Stifles of Dogs.

The cranial cruciate ligament rupture (CrCLR) is characterized by chronic inflammation and osteoarthritis (OA) of the stifle joint and extracellular matrix (ECM) degeneration of the ligament itself in dogs. Generally, OA may arise from chronic low-grade systemic inflammation. We assessed the possible relationship of inflammatory markers in the peripheral blood (PB) and synovial fluid (SF) of affected stifle joints in comparison to a control. Moreover, no study has shown the possible association between PB and SF levels of inflammatory markers in CrCLR stifles of dogs in veterinary medicine yet. We also evaluated components of ECM of CrCLR and finally compared the tibial plateau angle (TPA) and the anatomical-mechanical angle (AMA) between groups. Samples from PB and SF were examined for mRNA expression of interleukins, TNF-α and INF-γ. ECM components-collagen 1A1 and 3A1 and elastin-were examined for mRNA expression from SF. The level of relative expression for IL-1ß, IL-8 and IFN-γ was significantly increased in both PB and SF in CrCLR stifles as compared with the control. Collagens were also significantly increased in CrCLR stifles. TPA was not significantly different; however, the AMA angle significantly increased in the CrCLR group. Our results suggest a possible relationship between PB and SF levels of inflammatory markers in CrCLR stifles of dogs.

Study of bilateral elbow joint osteoarthritis treatment using conditioned medium from allogeneic adipose tissue-derived MSCs in Labrador retrievers.

Canine elbow dysplasia is a common cause of forelimb lameness in dogs and can lead to development of osteoarthritis (OA). A potential alternative to pain management is the use of a safe cell-free based therapy through trophic and paracrine factors of mesenchymal stem cells (MSCs). The aim of study was to identify the profile of selected mediators of potential clinical relevance in synovial fluid (SF) samples of dogs with elbow OA and analyse the range of motion (ROM) before and after cell-free MSCs-based treatment. In this study, conditioned medium from allogeneic canine adipose tissue - derived MSC (CM-AD-MSC) was prepared and administered into both elbow joints with OA in six Labrador retriever dogs (n = 6) on day 0 and 14 without creating a control group with a placebo. The SF of the elbow joints was analysed for the presence of several biomolecules (IL-6, IL-10, IL-8, IL-2, IL-12, TNF-αIFN-γ, MMP-3TIMP-1) before and after intraarticular applications of CM-AD-MSC. Kinematic analysis was used to assess the clinical effect of CM-AD-MSC. Analyses of SF and ROM were performed on days 0, 14 and 42. Concentration levels of MMP-3, TIMP-1, IL-6 and TNF-α in SF showed significant differences before and after the treatment (P < .05). There was a significant improvement in ROM between day 0 and 42 (P < .001). No severe adverse events were observed during the study. Results support the potential supportive effect of CM-AD-MSC as a noninvasive therapeutic tool for pain management of OA elbow joints in dogs.

Hypothermic treatment after computer-controlled compression in minipig: A preliminary report on the effect of epidural vs. direct spinal cord cooling.

The aim of the present study was to investigate the therapeutic efficacy of local hypothermia (beginning 30 min post-injury persisting for 5 h) on tissue preservation along the rostro-caudal axis of the spinal cord (3 cm cranially and caudally from the lesion site), and the prevention of injury-induced functional loss in a newly developed computer-controlled compression model in minipig (force of impact 18N at L3 level), which mimics severe spinal cord injury (SCI). Minipigs underwent SCI with two post-injury modifications (durotomy vs. intact dura mater) followed by hypothermia through a perfusion chamber with cold (epidural t≈15°C) saline, DMEM/F12 or enriched DMEM/F12 (SCI/durotomy group) and with room temperature (t≈24°C) saline (SCI-only group). Minipigs treated with post-SCI durotomy demonstrated slower development of spontaneous neurological improvement at the early postinjury time points, although the outcome at 9 weeks of survival did not differ significantly between the two SCI groups. Hypothermia with saline (t≈15°C) applied after SCI-durotomy improved white matter integrity in the dorsal and lateral columns in almost all rostro-caudal segments, whereas treatment with medium/enriched medium affected white matter integrity only in the rostral segments. Furthermore, regeneration of neurofilaments in the spinal cord after SCI-durotomy and hypothermic treatments indicated an important role of local saline hypothermia in the functional outcome. Although saline hypothermia (24°C) in the SCI-only group exhibited a profound histological outcome (regarding the gray and white matter integrity and the number of motoneurons) and neurofilament protection in general, none of the tested treatments resulted in significant improvement of neurological status. The findings suggest that clinically-proven medical treatments for SCI combined with early 5 h-long saline hypothermia treatment without opening the dural sac could be more beneficial for tissue preservation and neurological outcome compared with hypothermia applied after durotomy.

Neuroprotective effect of local hypothermia in a computer-controlled compression model in minipig: Correlation of tissue sparing along the rostro-caudal axis with neurological outcome.

This study investigated the neuroprotective efficacy of local hypothermia in a minipig model of spinal cord injury (SCI) induced by a computer-controlled impactor device. The tissue integrity observed at the injury epicenter, and up to 3 cm cranially and caudally from the lesion site correlated with motor function. A computer-controlled device produced contusion lesions at L3 level with two different degrees of tissue sparing, depending upon pre-set impact parameters (8N- and 15N-force impact). Hypothermia with cold (4°C) saline or Dulbecco's modified Eagle's medium (DMEM)/F12 culture medium was applied 30 min after SCI (for 5 h) via a perfusion chamber (flow 2 ml/min). After saline hypothermia, the 8N-SCI group achieved faster recovery of hind limb function and the ability to walk from one to three steps at nine weeks in comparison with non-treated animals. Such improvements were not observed in saline-treated animals subjected to more severe 15N-SCI or in the group treated with DMEM/F12 medium. It was demonstrated that the tissue preservation in the cranial and caudal segments immediately adjacent to the lesion, and neurofilament protection in the lateral columns may be essential for modulation of the key spinal microcircuits leading to a functional outcome. Tissue sparing observed only in the caudal sections, even though significant, was not sufficient for functional improvement in the 15N-SCI model.

Comparison of Radiographic Measurements of the Femur in Yorkshire Terriers with and without Medial Patellar Luxation.

OBJECTIVE: This article aimed to compare measurements of anatomical angles of the femurs, based on radiography, in Yorkshire Terriers without or with various grades of medial patellar luxation (MPL) based on radiography. METHODS: The skeletally mature Yorkshire Terrier dogs without MPL and with various grades of MPL were included in this prospective study. Cases with other orthopaedic disorders were excluded. For inclusion of the dog, it was required that standardized digital radiographs of both femurs in craniocaudal and axial directions were available. Measurements of the anatomical lateral proximal femoral angle, anatomical lateral distal femoral angle, femoral varus angle, anteversion angle and femoral inclination angle were performed. RESULTS: Forty-two Yorkshire Terriers (84 hindlimbs) were included in the study. They were divided into five groups according to grade of MPL as healthy (n = 12), grade I (n = 9), grade II (n = 44), grade III (n = 10) and grade IV (n = 9) dogs. The anatomical lateral proximal femoral angle and anteversion angle were significantly lower in dogs with grade IV MPL, while anatomical lateral distal femoral angle and femoral varus angle were significantly higher. The femoral inclination angle values did not differ significantly. CLINICAL SIGNIFICANCE: Yorkshire Terriers affected with grade IV MPL had severe femoral deformities or femoral varus and external rotation of the distal femur. Reference range can be used as an aid in diagnosis, determining indications and surgical planning for corrective osteotomy.

A Preliminary Study Comparing Microfracture and Local Adherent Transplantation of Autologous Adipose-Derived Stem Cells Followed by Intraarticular Injection of Platelet-Rich Plasma for the Treatment of Chondral Defects in Rabbits.

Objective This study aimed to compare microfracture and application of adipose-derived stem cells (ADSCs) by local adherent technique enhanced by platelet-rich plasma (PRP) to provide a new approach for the repair of cartilage defect. Design Full-thickness cylindrical defects were created in the medial femoral condyle in 9 New Zealand White rabbits (5 months old, 4.65 ± 0.20 kg). Two groups of rabbits ( n = 3) were either treated with ADSCs (Group 1) or the microfracture technique (Group 2) following intraarticular injection of PRP 3 times in weekly intervals. Rabbits in control group ( n = 3) remained untreated. The outcome was assessed macroscopically, histologically, and immunohistochemically. Results At the end of week 12, Group 1 showed better defect filling compared with Group 2. Specimens treated with the combination of ADSCs and PRP exhibited significant differences from the other groups in all criteria of International Cartilage Repair Society macroscopic scoring system. Conclusions Intraarticular injection of autologous PRP in combination with transplantation of autologous ADSCs by local adherent technique enhances the quality of cartilage defect repair with better results in comparison with microfracture surgery in a rabbit model.

The use of native fluorescence analysis of synovial fluid in the diagnosis of medial compartment disease in medium- and large-breed dogs.

We assumed that proteins are most likely responsible for synovial fluid fluorescence and that changes detected in fluorescence intensity are most likely the result of changes in the concentration of fluorescent proteins. Synchronous fluorescent matrices from synovial fluid samples were measured in the excitation wavelength range of 200-350 nm using a luminescence spectrophotometer. The synchronous matrix of synovial fluid consists of 2 dominant fluorescent centers (F1 and F2) in the ultraviolet region. The fluorescence intensities of both centers were significantly higher in pathological samples, with p = 0.001 (a 59% increase of the median value) for the F1 center and p = 0.002 (a 52% increase of the median value) for the F2 center. Receiver operating characteristic analysis confirmed that synovial fluid autofluorescence is a significant predictor of medial compartment disease in dogs, with the area under the curve at 0.776 (F1) and 0.778 (F2). We did not detect any differences in the autofluorescence of synovial fluid between male and female, or any breed-based changes. No position changes of fluorescent centers were recorded in the synovial fluid in diseased dogs compared with healthy dogs. The synovial fluid metabolic fingerprint of canine patients with medial compartment disease differed from that of healthy dogs. Our study demonstrated the feasibility of synovial fluid fingerprinting to identify disease-specific profiles of synovial fluid metabolites.

Histological Assessment of a Combined Low-Level Laser/Light-Emitting Diode Therapy (685 nm/470 nm) for Sutured Skin Incisions in a Porcine Model: A Short Report.

OBJECTIVE: The aim of our study was to evaluate, from a histological point of view, the effect of photobiomodulation (PBM) with combined low-level laser therapy (LLLT)/light- emitting diode (LED) on porcine skin wound healing. BACKGROUND DATA: Most LLLT/LED wound healing studies have been performed on various types of rat models, with their inherent limitations. Minipigs are evolutionary and physiologically closer to humans than rats. MATERIALS AND METHODS: With the animals under general anesthesia, one full-thickness skin incision was performed on the back of each minipig (n = 10) and immediately closed using simple interrupted percutaneous sutures. The minipigs were randomly allocated into two groups: a PBM-treated group (LLLT λ = 685 nm, LED λ = 470 nm, both light sources producing power densities at 0.008 W/cm2; each light source delivering total daily doses of 3.36 J/cm2) and a sham-irradiated control group. Half of the animals in each group were killed on postoperative day 3, and the other half were killed on the postoperative day 7, and samples were removed for histological examination. RESULTS: Combined red and blue PBM accelerated the process of re-epithelization and formation of cross-linked collagen fibers compared with sham irradiated control wounds. CONCLUSIONS: Our results demonstrate that the current dose of combined red and blue PBM improves the healing of sutured skin incisions in minipigs.

No Sexual Dimorphism Detected in Digit Ratios of the Fire Salamander (Salamandra salamandra).

It has been proposed that digit ratio may be used as a biomarker of early developmental effects. Specifically, the second-to-fourth digit ratio (2D:4D) has been linked to the effects of sex hormones and their receptor genes, but other digit ratios have also been investigated. Across taxa, patterns of sexual dimorphism in digit ratios are ambiguous and a scarcity of studies in basal tetrapods makes it difficult to understand how ratios have evolved. Here, we focus on examining sex differences in digit ratios (2D:3D, 2D:4D, and 3D:4D) in a common amphibian, the fire salamander (Salamandra salamandra). We used graphic software to measure soft tissue digit length and digit bone length from X-rays. We found a nonsignificant tendency in males to have a lower 2D:3D than females; however, no sexual differences were detected in the other ratios. We discuss our results in the context of other studies of digit ratios, and how sex determination systems, as well as other factors, might impact patterns of sexual dimorphism, particularly in reptiles and in amphibians. Our findings suggest that caution is needed when using digit ratios as a potential indicator of prenatal hormonal effects in amphibians and highlight the need for more comparative studies to elucidate the evolutionary and genetic mechanisms implicated in sexually dimorphic patterns across taxonomic groups.

A preliminary trial of the sedation induced by intranasal administration of midazolam alone or in combination with dexmedetomidine and reversal by atipamezole for a short-term immobilization in pigeons.

OBJECTIVE: To assess the sedative and immobilization effect of intranasal administration (INS) of midazolam (MID) without or with INS dexmedetomidine (DXM), and some physiological changes induced by the drugs. The ability of INS atipamezole to reverse the DXM component was also assessed. STUDY DESIGN: Prospective 'blinded' experimental study. ANIMALS: In total, 15 pigeons. METHODS: Pigeons were sedated by INS MID alone at a dose of 5 mg kg(-1) (group MID, n = 6) or in combination with INS DXM at a dose 80 µg kg(-1) (group MID-DXM, n = 6). Measurements were made of heart rate (HR), respiratory rate (fR ) and cloacal temperature (CT). The degree of sedation was assessed at 15 minutes prior to, immediately after, and at intervals until 100 minutes after drug administrations. Following MID-DXM, INS atipamezole (250 µg kg(-1) ) was administered and the same indices measured 5 and 10 minutes later. RESULTS: MID had no effect on HR and fR , and although CT decreased, it remained within physiological range. MID-DXM caused significant falls in HR, fR and CT that persisted until the end of sedation. Atipamezole antagonized sedation and cardiorespiratory side effects of MID-DXM within 10 minutes of application. In addition, for MID compared to MID-DXM, the lowest sedation scores [10 (7-14) and 10.5 (5-14) versus 2 (1-4) and 2 (1-5)] were achieved in the 10th and 20th minute versus the 20th and 30th minute of the sedation, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: MID alone, given INS had minimal side effects on vital functions but caused inadequate immobilization of pigeons for restraint in dorsal recumbency. MID-DXM caused an effective degree of immobilization from 20 to 30 minutes after administration, at which time birds tolerated postural changes without resistance. Atipamezole antagonized both side effects and sedation, but complete recovery had not occurred within 10 minutes after its application.

Isolation, cultivation and characterisation of pigeon osteoblasts seeded on xenogeneic demineralised cancellous bone scaffold for bone grafting.

Avian osteoblasts have been isolated particularly from chicken embryo, but data about other functional tissue sources of adult avian osteoblast precursors are missing. The method of preparation of pigeon osteoblasts is described in this study. We demonstrate that pigeon cancellous bone derived osteoblasts have particular proliferative capacity in vitro in comparison to mammalian species and developed endogenous ALP. Calcium deposits formation in vitro was confirmed by alizarin red staining. Only a few studies have attempted to investigate bone grafting and treatment of bone loss in birds. Lack of autologous bone grafts in birds has prompted investigation into the use of avian xenografts for bone augmentation. Here we present a method of xenografting of ostrich demineralised cancellous bone scaffold seeded with allogeneic adult pigeon osteoblasts. Ostrich demineralised cancellous bone scaffold supported proliferation of pigeon osteoblasts during two weeks of co - cultivation in vitro. Scanning electron microscopy demonstrated homogeneous adult pigeon osteoblasts attachment and distribution on the surface of xenogeneic ostrich demineralised cancellous bone. Our preliminary in vitro results indicate that demineralised cancellous bone from ostrich tibia could provide an effective biological support for growth and proliferation of allogeneic osteoblasts derived from cancellous bone of pigeons.

Oral administration of inosine promotes recovery after experimental spinal cord injury in rat.

Inosine, a purine nucleoside, is one of the novel substances, which can preserve the neuronal and glial viability and stimulate intact neurons to extend axons. We, herein, evaluated the effect of oral inosine treatment on spinal cord injury (SCI) recovery by means of locomotor and bladder function, quantification of neurons and spinal cord tissue sparing. Rats after compression SCI were divided into groups-SCI-Aqua and SCI-Inosine (daily application of aqua for injection or inosine)-locomotion of hind limbs (BBB score) and urinary bladder function were evaluated from day 1 to 28 after SCI. The neuronal profile was determined by immunohistochemistry with NeuN antibodies and tissue sparing by Luxol fast blue staining method. SCI affected the functional movement of hind limbs in both groups with gradual improvement (increased BBB score) during survival. However, we found a significant difference in BBB score and recovery of bladder function between SCI-Aqua and SCI-Inosine groups during the second week of survival following SCI. In addition, the number of NeuN positive cells and percentage of tissue sparing was also significantly higher in SCI-Inosine group when compared with the SCI-Aqua group. Daily oral administration of inosine after SCI throughout the survival was beneficial for locomotion and micturition, neuronal survival and tissue sparing. This indicates that inosine may represent one of the co-stimulatory factors for treatment strategies to promote neuronal plasticity after SCI.

Experimental anterior lens capsule transplantation for chronic corneal ulcers-Bowman's layer replacement?

PURPOSE: This study was designed to measure the value of allografted anterior lens capsule in the reepithelialization of recurrent corneal ulcers. METHODS: Mechanical ulcers of uniform size were created with a 6-mm corneal trephine in both eyes of four Chinchilla male rabbits at one-third corneal depth. Following initial epithelial regrowth, an identical injury was created in the same area of each cornea a second time. In four eyes (treated group), an anterior lens capsule from a healthy donor rabbit was sutured into the ulcer bed followed by antibiotic/steroid drops three times daily for 1 week. The remaining four control eyes were allowed to heal without surgical intervention using the same antibiotic/steroid drops only. Slit-lamp examination and histopathology findings were recorded over a 6-month follow-up period. RESULTS: The four treated eyes reepithelialized after the second injury at a faster rate than the control group eyes and had a lower percentage of corneal opacification at all follow-up exam dates. Histopathology revealed normal epithelium overlying the transplanted anterior lens capsule and no infiltration of inflammatory cells in the subepithelial stroma. CONCLUSIONS: In experimental conditions, allotransplantation of anterior lens capsule speeds up reepithelialization after repeated mechanical trauma to the cornea. To our knowledge, this is the first experimental transplantation of an anterior lens capsule to create an artificial Bowman's layer in recurrent corneal ulcerations to aid reepithelialization and minimize corneal scarring.

Displacement of maxillary premolar teeth in a filly.

Abnormalities of dental development such as hypoplasia of cementum, oligodontia, polydontia, enamel hypoplasia, brachygnathia, and prognathia are quite common in the horse. Abnormalities of eruption are less common and often associated with trauma. This case report describes the diagnosis and therapy for pre-eruption displacement of the maxillary left third and fourth premolar teeth resulting from previous facial trauma in a 1.5-year-old filly.