Two Amnion-Derived Mesenchymal Stem-Cells Injections to Osteoarthritic Elbows in Dogs-Pilot Study.

The aim of the study was to investigate the potential of cell-based regenerative therapy for elbow joints affected by osteoarthritis. Interest was focused on two intra-articular applications of amnion-derived mesenchymal stem cells (A-MSCs) to a group of different breeds of dogs with elbow osteoarthritis (13 joints). Two injections were performed 14 days apart. We evaluated synovial fluid biomarkers, such as IFN-γ, IL-6, IL-15, IL-10, MCP-1, TNF-α, and GM-CSF, by multiplex fluorescent micro-bead immunoassay in the treated group of elbows (n = 13) (day 0, day 14, and day 28) and in the control group of elbows (n = 9). Kinematic gait analysis determined the joint range of motion (ROM) before and after each A-MSCs application. Kinematic gait analysis was performed on day 0, day 14, and day 28. Kinematic gait analysis pointed out improvement in the average range of motion of elbow joints from day 0 (38.45 ± 5.74°), day 14 (41.7 ± 6.04°), and day 28 (44.78 ± 4.69°) with statistical significance (p < 0.05) in nine elbows. Correlation analyses proved statistical significance (p < 0.05) in associations between ROM (day 0, day 14, and day 28) and IFN-γ, IL-6, IL-15, MCP-1, TNF-α, and GM-CSF concentrations (day 0, day 14, and day 28). IFN-γ, IL-6, IL-15, MCP-1, GM-CSF, and TNF- α showed negative correlation with ROM at day 0, day 14, and day 28, while IL-10 demonstrated positive correlation with ROM. As a consequence of A-MSC application to the elbow joint, we detected a statistically significant (p < 0.05) decrease in concentration levels between day 0 and day 28 for IFN-γ, IL-6, and TNF-α and statistically significant increase for IL-10. Statistical significance (p < 0.05) was detected in TNF-α, IFN-γ, and GM-CSF concentrations between day 14 and the control group as well as at day 28 and the control group. IL-6 concentrations showed statistical significance (p < 0.05) between day 14 and the control group.

Ultrasound Evaluation of Extracranial Cerebral Circulation (The Common, External and Internal Carotid Artery) in Different Breeds of Dogs.

Noninvasive Doppler ultrasonography (US) examination is a standard method for the clinical evaluation of the carotid arteries. Extracranial cerebral circulation includes the common carotid artery (CCA), the external carotid artery (ECA), and the extracranial part of the internal carotid artery (ICA). The present study was conducted with the objective of assessing physiological normative values and describing the appearance of spectral waveforms of extracranial arteries in 104 healthy dogs of eight breeds divided into four groups according to weight. We also focused on searching for correlations for carotid blood velocities with the resistive index (RI), body weight and diameter, and differences between observers and the influence of sex in the calculations of Doppler parameters. In the evaluated breeds, significant differences were found in the velocity of peak systolic velocity (PSV) and end diastolic velocity (EDV). There was a strong correlation between body weight and peak systolic velocity, the RI index and CCA diameter. The intra-observer agreement for the PSV and EDV parameters in each vessel was considered excellent reliability, and overall interobserver agreement showed very good reliability. This study could improve the descriptions of physiological values and waveforms recorded in carotid arteries. Defining the physiological values of velocity and the resistive index (RI) helps in the easier identification of pathology and diagnosis of disease. Our results may be used for further studies of vascular diseases in veterinary medicine that correlate with the pathology of neurological disorders of ischemic origin, further in thromboembolism, oncologic disease and degenerative, proliferative and inflammatory diseases of the arteries that lead to stenosis.

Retrospective analysis of tracheal hypoplasia in brachycephalic and non-brachycephalic dogs: Inter- and intra-observer agreement of measurements.

This retrospective study was undertaken on the records of intraluminal diameter of the trachea in 185 dogs, in which hypoplasia of the trachea had been suspected. The relative size of the trachea was measured using the tracheal diameter (TD), thoracic inlet distance (TI), thoracic tracheal diameter (TT) and the width of the third rib (3R), expressed as ratios TD:TI and TT:3R. Thirty-five dogs were diagnosed as having tracheal hypoplasia. Bulldogs and non-bulldog brachycephalic dogs had significantly smaller measured trachea diameters compared to the predicted values calculated on the basis of their body weight. Radiographs of each dog were investigated by four observers. Inter- and intra-observer reliability (ICCinter, ICCintra) was based on the measurements taken by four observers to evaluate the reproducibility of the protocol. There was a good ICCinter (0.8) and ICCintra (0.89) agreement. Craniocaudal tangential radiographs, centred on the cranial thoracic aperture, did not show a significant difference in tracheal diameter measurements compared to the right lateral radiographs. In conclusion, our findings indicate that bulldogs and non-bulldog brachycephalic dogs have smaller tracheal diameters than non-brachycephalic dogs.

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.

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.

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.

The influence of omega-3 polyunsaturated fatty acids (omega-3 pufa) on lactobacilli adhesion to the intestinal mucosa and on immunity in gnotobiotic piglets.

The effect of application of omega-3 polyunsaturated fatty acids (omega-3 PUFA) on intestinal colonization by Lactobacillus paracasei and on cellular immunity has been investigated in gnotobiotic pigs. The administration of polyunsaturated fatty acids positively affected the adhesion of Lactobacillus paracasei to the jejunal mucosa of gnotobiotic piglets. When compared to the control group, the number of Lactobacillus paracasei adhering to the jejunal mucosa was by 12% higher in piglets of the experimental group (5.10 log 10/cm2 vs. 4.55 log 10/cm2). The respective counts of Lactobacillus paracasei adhering to the ileal and colonic mucosa of 28 day old gnotobiotic piglets reached 4.45 and 5.05 log 10/cm2 in group C and 4.44 and 4.95 log 10/cm2 in group E. Omega-3 PUFA supplementation increased the phagocytic activity of neutrophils by almost 100% on day 28 of life as well as the subpopulations of lymphocytes (CD8) in the peripheral blood of germ-free piglets on day 21 of life. Our results indicate that the action of probiotics in the gut may be modulated by dietary PUFA. The stimulatory effect of PUFA upon adhesion of lactobacilli could be used for enhancing the effectiveness of probiotics in inhibiting digestive tract pathogens.