Canine Bone Research Articles

Human placental hydrolysate promotes the long-term culture of hepatocyte-like cells derived from canine bone marrow.

Long-term culture of canine artificial hepatocytes has not been established. We hypothesized that human placental hydrolysate (hPH) may support the long-term culture of differentiated hepatocyte-like cells. Canine bone marrow cells were cultured using modified hepatocyte growth medium supplemented with hPH. Quantitative reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemical analysis for albumin, qualitative RT-PCR for cytochrome P450 1A1 (CYP1A1), hepatocyte growth factor (HGF), Cytokeratin 7 (CK7), CD90, CD44, and CD34, and functional analyses of CYP450 activity and low-density lipoprotein (LDL) uptake were performed. Cultured hepatocyte-like cells were able to maintain hepatocyte characteristics, including morphology, albumin synthesis, CYP450 activity, and LDL uptake for 80 days. Thus, hPH may be a potential facilitator for the long-term culture of hepatocyte-like cells. Clinicopathologically, this culture protocol of artificial hepatocytes will contribute to liver function evaluation.

3D anatomical model for teaching canine lumbosacral epidural anesthesia.

Purpose To develop a 3D anatomical model for teaching canine epidural anesthesia (3DMEA) and to assess its efficacy for teaching and learning prior to the use of live animals.Methods The creation of 3DMEA was based on 3D optical scanning and 3D printing of canine bone pieces of the fifth to the seventh lumbar vertebrae, sacrum and pelvis. A total of 20 male dogs were scheduled for castration. 20 veterinary students watched a video showing epidural anesthesia in dogs before the clinical attempt and were assigned to control or 3DMEA groups. Students in the 3DMEA group trained in the model after the video. For the clinical trial, the epidural procedure was performed by students under the veterinary supervision. When observed the absence of response to nociceptive stimuli, the epidural was considered successful. Then, all students answered a questionnaire evaluating the main difficulty founded in the technique and its degree of difficulty.Results The 3DMEA group reported a lower degree of difficulty to perform the epidural anesthesia technique when compared with the control group (p=0.0037). The 3DMEA reproduced the anatomical structures, allowing the perception of the distance of needle in relation to the iliac prominences during epidural anesthesia. Its mobility allowed simulation of the animal in standing position and sternal recumbency.Conclusion The use of 3DMEA demonstrated greater efficacy in the execution of the technique, being effective in the teaching and learning process before the epidural anesthesia in live animals.

Medical infrared thermal imaging of canine appendicular bone neoplasia

BackgroundMedical infrared thermal imaging (MITI) is a noninvasive imaging modality used in veterinary medicine as a screening tool for musculoskeletal and neurological disease processes. An infrared camera measures the surface body heat and produces a color map that represents the heat distribution. Local trauma or disease can impair the autonomic nervous system, which leads to changes in the local dermal microcirculation and subsequent alteration of surface body heat. Disruption of autonomic flow to the cutaneous vasculature at deeper levels can also result in asymmetric thermographic results. The purpose of this study was to evaluate surface temperature differences between limbs affected by bone neoplasia and their normal contralateral limbs.ResultsA statistically significant difference in average temperature was noted between regions of interest of the two groups (paired difference: 0.53 C° ± 0.14; P = 0.0005). In addition, pattern recognition analysis yielded a 75–100% success rate in lesion identification.ConclusionsSignificant alterations noted with average temperature and thermographic patterns indicate that MITI can document discernible changes associated with the presence of canine appendicular bone tumors. While MITI cannot be used as the sole diagnostic tool for bone cancer, it can be used as a screening modality and may be applicable in early detection of cancer.

Guidelines for the Execution of True Spherical Osteotomies Using a Modified Dome Blade Design.

The purpose of this study was to explore the trigonometric principles of the spherical osteotomy, establish guidelines for its application and test the guidelines on bone models using a new blade design. We propose a new rule of osteotomies incorporating the outlined geometric principles, and applicable to the use of spherical cuts in veterinary orthopaedic surgery. The trigonometric principles for the execution of neutral, closing and opening spherical osteotomies were explored in silico. A modification of the existing commercially available dome blade was designed and manufactured such that it facilitated the performance of spherical osteotomy with a minimized blade radius. A pilot study was performed whereby the modified dome blade was used to create spherical osteotomy in canine radial bone models. The surfaces of the osteotomy models were laser-scanned using a three-dimensional (3D) scanner; the resultant scans were imported into and analysed using a commercial 3D analysis software. The accuracy of osteotomy execution was measured as the distance between the targeted centre of osteotomy and the actual centre of osteotomy as found on the 3D scans. By utilizing the geometric principles of spherical osteotomy, an accurate osteotomy position was achieved. The centre of the spherical cut performed on bone models was confirmed to be within 5% tolerance of the location as planned in silico demonstrating the accurate and relevant clinical application of geometric principles. The trigonometric guidelines for the execution of spherical osteotomy can be applied in a pre-clinical environment with accuracy. The new guidelines combined with the proposed new rule for spherical osteotomy utilizing the new blade design are translatable into clinical application, permitting the surgeon to accurately plan osteotomy application while mitigating the significant loss of bone-to-bone contact during correction of torsional deformities inherent in the principles of dome osteotomy use.

Effect of cryopreservation on therapeutic potential of canine bone marrow derived mesenchymal stem cells augmented mesh scaffold for wound healing in guinea pig

The objective of this study was to compare the therapeutic potential of canine bone marrow derived mesenchymal stem cells (BM MSCs) augmented mesh scaffold for wound healing potential in guinea pig before and after cryopreservation. Bone marrow aspirate was obtained from healthy dogs and culture was expanded in vitro. MSCs augmented mesh scaffold were cryopreserved for 30 days and then used for therapeutic purposes. Both fresh and frozen thaw MSCs augmented mesh scaffold along with fresh MSCs were used for therapeutic purposes in guinea pig. No significant (P > 0.05) difference was observed in population doubling time (PDT) among fresh and frozen thawed BM MSCs. Both fresh and frozen thawed BM MSCs expressed cell surface markers (CD73, CD90, and CD105), and did not express CD34 as was confirmed by Immunocytochemistry and Real-Time Polymerase Chain Reaction. The fresh and frozen thawed BM MSCs successfully differentiated into osteogenic, chondrogenic and adipogenic lineages. Therapeutic results revealed that the percent wound contraction on day 14 was more than 65 % for the mesh augmented with MSCs as well as freshly injected MSCs group as against 33–34 % in the control group. Healed wound quality parameters viz. surface epithelium, neovascularization, and collagen characteristics were better for the mesh augmented with MSCs as well as freshly injected MSCs group compared to the control group. No significant difference was noted among fresh and frozen thawed BM MSCs group and fresh MSCs injected group. Thus, it is concluded from this study that canine BM MSCs augmented mesh scaffold both fresh and frozen thaw can be used for quality wound healing.

In vitro proliferation and differentiation of canine bone marrow derived mesenchymal stem cells over hydroxyl functionalized CNT substrates

Nanotopography of culture substrate acts as a positive cue in cell-biomaterial based tissue regeneration. Considering the potentiality of carbon nanotubes (CNTs) this study was designed to evaluate its two functionalized form by an in vitro culture condition using canine mesenchymal stem cells as cellular model. Cells were isolated and its behaviour, proliferation and differentiation processes were elucidated onto CNT substrates. Beside the variations in cellular behaviour it was remarkably noted that even though proliferation was reduced but osteogenic and chondrogenic differentiation was enhanced over multi-walled CNTs, whereas neuronal differentiation was better supported by single walled CNTs as evidenced by our cytochemical, immunocytochemical, gene expression and flow cytometry assays. The former one was noticed more cytocompatible by our different apoptosis studies. The outcome of these experiments collectively indicated that hydroxylated functionalized CNTs could be a potential scaffold constituent for future experimentations as well as for the application in regenerative medicine.

Effect of orthopedic implants on canine long bone compression stiffness: a combined experimental and computational approach.

Osteosynthesis for canine long bones is a complex process requiring knowledge of biology, surgical techniques and (bio)mechanical principles. Subject-specific finite element analysis constitutes a promising tool to evaluate the effect of surgical intervention on the global properties of a bone-implant construct, but suffers from a lack of validation. In this study, the biomechanical behavior of 10 canine humeri was compared before and after creation of a 10 mm bone defect stabilized with an eight-hole locking compression plate (Synthes®) and two locking screws on each fragment. The response under compression of both intact and plated samples was measured experimentally and reproduced with a finite element model. The experimental stiffness ratio between plated and intact bone was equal to 0.39 ± 0.06. A subject-specific finite element analysis including density-dependent elasto-plastic material properties for canine bone and automatic generation of orthopedic implants was then conducted to recover these experimental results. The stiffness of intact and plated samples could be predicted, with no significant differences with experimental data. The simulated stiffness ratio between plated and intact canine bone was equal to 0.43 ± 0.03. This study constitutes a first step toward the building of a virtual database of pre-computed cases, aiming at helping the veterinary surgeons to make decisions regarding the most suited orthopedic solution for a given dog and a given fracture.

Real-time PCR to differentiate among Leishmania (Viannia) subgenus, Leishmania (Leishmania) infantum and Leishmania (Leishmania) amazonensis: Application on Brazilian clinical samples

Leishmaniasis is a complex disease caused by Leishmania species belonging to subgenera Leishmania and Viannia. In South America, L. (L.) infantum is considered the most important causative agent of visceral leishmaniasis, while L. (L.) amazonensis and Viannia subgenus species are responsible for the different cutaneous or mucocutaneous forms. In our previous work, we developed a diagnostic approach for Leishmania species discrimination based on two qPCRs (qPCR-ML and qPCR-ama) targeting the minicircle kDNA followed by melting analysis. This approach allowed to (i) differentiate the subgenera Leishmania and Viannia, and (ii) distinguish between L. (L.) infantum and L. (L.) amazonensis. The aim of this work was to demonstrate the applicability of the approach previously described, using human and canine clinical samples and strains from a Brazilian region, where L. (L.) infantum, L. (L.) amazonensis and Viannia subgenus species coexist. After validation on New World strains, the diagnostic approach was applied blindly to 36 canine clinical samples (peripheral blood and bone marrow) and 11 human clinical samples (peripheral blood and bone marrow). The sensitivity was 95.6% (95% confidence interval 77.3–100%) and 100% (95% confidence interval 76.9–100%) in the canine bone marrow samples and human (peripheral blood and bone marrow) samples, respectively, compared to conventional PCR assays. Concerning the Leishmania species identification, the conventional and qPCR-based methods showed kappa value of 0.876 (95% confidence interval 0.638–1.000), indicating good agreement. Therefore, this approach proved to be useful in both veterinary and human clinical context in regions co-endemic for L. (L.) infantum, L. (L.) amazonensis, and Viannia subgenus, helping to provide rapid diagnosis and to allow studies of species distribution.

Simvastatin enhances proliferation and pluripotent gene expression by canine bone marrow-derived mesenchymal stem cells (cBM-MSCs) in vitro

Establishing the intervention to enhance proliferation and differentiation potential is crucial for the clinical translation of stem cell-based therapy. In this study, the effects of simvastatin on these regards were explored. Canine bone marrow-derived mesenchymal stem cells (cBM-MSCs) were treated with 4 doses of simvastatin, 0.1, 1, 10, and 100 nM. Simvastatin in low-dose range, 0.1 and 1 nM, enhanced dose-dependent cell proliferation at day 5 and 7. Exploration of the mechanisms revealed that simvastatin in low-dose range dose-dependently upregulated sets of cell cycle regulators, Cyclin D1 and Cyclin D2; proliferation marker, Ki-67; and anti-apoptotic gene; Bcl-2. Interestingly, pluripotent markers, Rex1 and Oct4, were dramatically increased upon the low-dose treatment. Contrastingly, treatment with high-dose simvastatin suppressed the expression of those genes. Thus, the results suggested beneficial effects of simvastatin on cBM-MSCs proliferation and expansion. Further study regarding differentiation potential and underlying mechanisms will accelerate the clinical application of the molecule on veterinary stem cell-based therapy.

Investigation of effective intrusion and extrusion force for maxillary canine using finite element analysis

Orthodontic tooth movement is mainly regulated by the biomechanical responses of loaded periodontal ligament (PDL). We investigated the effective intervals of orthodontic force in pure maxillary canine intrusion and extrusion referring to PDL hydrostatic stress and logarithmic strain. Finite element analysis (FEA) models, including a maxillary canine, PDL and alveolar bone, were constructed based on computed tomography (CT) images of a patient. The material properties of alveolar bone were non-uniformly defined using HU values of CT images; PDL was assumed to be a hyperelastic–viscoelastic material. The compressive stress and tensile stress ranging from 0.47 to 12.8 kPa and 18.8 to 51.2 kPa, respectively, were identified as effective for tooth movement; a strain 0.24% was identified as the lower limit of effective strain. The stress/strain distributions within PDL were acquired in canine intrusion and extrusion using FEA; root apex was the main force-bearing area in intrusion–extrusion movements and was more prone to resorption. Owing to the distinction of PDL biomechanical responses to compression and tension, the effective interval of orthodontic force was substantially lower in canine intrusion (80–90 g) than in canine extrusion (230–260 g). A larger magnitude of force remained applicable in canine extrusion. This study revised and complemented orthodontic biomechanical behaviours of tooth movement with intrusive–extrusive force and could further help optimize orthodontic treatment.

Canine Bone Marrow-derived Mesenchymal Stem Cells: Genomics, Proteomics and Functional Analyses of Paracrine Factors

Adult stem cells have become prominent candidates for treating various diseases in veterinary practice. The main goal of our study was therefore to provide a comprehensive study of canine bone marrow-derived mesenchymal stem cells (BMMSC) and conditioned media, isolated from healthy adult dogs of different breeds. Under well-defined standardized isolation protocols, the multipotent differentiation and specific surface markers of BMMSC were supplemented with their gene expression, proteomic profile, and their biological function. The presented data confirm that canine BMMSC express important genes for differentiation toward osteo-, chondro-, and tendo-genic directions, but also genes associated with angiogenic, neurotrophic, and immunomodulatory properties. Furthermore, using proteome profiling, we identify for the first time the dynamic release of various bioactive molecules, such as transcription and translation factors and osteogenic, growth, angiogenic, and neurotrophic factors from canine BMMSC conditioned medium. Importantly, the relevant genes were linked to their proteins as detected in the conditioned medium and further associated with angiogenic activity in chorioallantoic membrane (CAM) assay. In this way, we show that the canine BMMSC release a variety of bioactive molecules, revealing a strong paracrine component that may possess therapeutic potential in various pathologies. However, extensive experimental or preclinical trials testing canine sources need to be performed in order to better understand their paracrine action, which may lead to novel therapeutic strategies in veterinary medicine.

Platelet-rich plasma affects the proliferation of canine bone marrow-derived mesenchymal stromal cells in vitro

BackgroundReported efficacy of platelet-rich plasma (PRP) in regenerative medicine is contradictory. We validated the effects of PRP on proliferation of canine bone marrow-derived multipotent mesenchymal stromal cells (K9BMMSCs) in vitro. PRP was extracted from blood of six dogs with osteoarthritis. K9BMMSCs were established from bone marrow and characterized for CD90 and CD19 expression by immunocytochemistry. Effects of PRP concentrations on viability of matching autologous K9BMMSCs were validated using MTS assay.ResultsPositive CD90 and negative CD19 expression confirmed MSC origin. PRP at 40% volume/volume concentration increased, while PRP at 80 and 100% v/v concentrations suppressed viability of tested K9BMMSCs.ConclusionPRP concentration plays an important role in K9BMMSCs viability, which could affect tissue repairs in vivo.

Analysis of Mechanical Properties and Mechanical Anisotropy in Canine Bone Tissues of Various Ages.

The effect of age on mechanical behavior and microstructure anisotropy of bone is often ignored by researchers engaged in the study of biomechanics. The objective of our study was to determine the variations in mechanical properties of canine femoral cortical bone with age and the mechanical anisotropy between the longitudinal and transverse directions. Twelve beagles divided into three age groups (6, 12, and 36 months) were sacrificed and all femurs were extracted. The longitudinal and transverse samples of cortical bone were harvested from three regions of diaphysis (proximal, central, and distal). A nanoindentation technique was used for simultaneously measuring force and displacement of a diamond tip pressed 2000nm into the hydrated bone tissue. An elastic modulus was calculated from the unloading curve with an assumed Poisson ratio of 0.3, while hardness was defined as the maximal force divided by the corresponding contact area. The mechanical properties of cortical bone were determined from 852 indents on two orthogonal cross-sectional surfaces. Mean elastic modulus ranged from 7.56±0.32 GPa up to 21.56±2.35 GPa, while mean hardness ranged from 0.28±0.057 GPa up to 0.84±0.072 GPa. Mechanical properties of canine femoral cortical bone tended to increase with age, but the magnitudes of these increase for each region might be different. The longitudinal mechanical properties were significantly higher than that of transverse direction (P<0.01). A significant anisotropy was found in the mechanical properties while there was no significant correlation between the two orthogonal directions in each age group (r2<0.3). Beyond that, the longitudinal mechanical properties of the distal region in each age group were lower than the proximal and central regions. Hence, mechanical properties in nanostructure of bone tissue must differ mainly among age, sample direction, anatomical sites, and individuals. These results may help a number of researchers develop more accurate constitutive micromechanics models of bone tissue in future studies.

Three-dimensional finite element analysis to evaluate biomechanical effects of different alveolar decortication approaches on rate of canine retraction

The aim of this study was to compare different corticotomy approaches and determine their biomechanical effects on rate of canine displacement when compared to conventional orthodontics. Three-dimensional Finite Element Models with conventional non-corticotomy approach (model 1) and three corticotomy approaches ensuing buccal and palatal vertical cuts (model 2), interseptal bone reduction (model 3), buccal vertical cuts (model 4) were fabricated. Displacement of the canine and von Mises stresses in the canine and trabecular bone were calculated and compared under a distal retraction force of 1.5N. The maximum displacement of canine with minimum anchorage loss was seen in model 3 followed by model 2, model 4 and model 1. The maximum equivalent (von Mises) stress was concentrated mainly on the distal side of canine in model 3 and had a uniform distribution of stresses on entire root surface. Corticotomy approaches effectively accelerated maxillary canine retraction, exhibiting twice the rate of canine movement with minimum anchorage loss when compared to non-corticotomy approach. Corticotomy with interseptal bone reduction was most effective in terms of canine displacement and stress distribution.

6-Bromoindirubin-3'-oxime promotes osteogenic differentiation of canine BMSCs through inhibition of GSK3β activity and activation of the Wnt/β-catenin signaling pathway.

This study aimed to investigate how 6-bromoindirubin-3'-oxime (BIO) increases the osteogenic differentiation of canine bone mesenchymal stem cells (BMSCs) and the role of the Wnt/β-catenin signaling pathway in this process. We mimicked the effect of Wnt by adding BIO to the culture medium of BMSCs and examined whether canonical Wnt signaling positively affects the differentiation of these cells into osteoblasts. Canine BMSCs were cultured with 0.5 and 1.0 μM BIO under osteogenic conditions and then differentiation markers were investigated. It was found that BIO significantly increased the activity of alkaline phosphatase (ALP), the number of ALP-positive cells, the mineralization level and calcium deposits. Moreover, cells cultured with 0.5 and 1.0 μM BIO exhibited detectable β-catenin expression in their nuclei, and showed upregulated β-catenin and glycogen synthase kinase 3 beta(GSK3β) phosphorylation compared to untreated cells. In addition, BIO enhanced the mRNA expression of osteoblast differentiation markers such as ALP, runt-related transcription factor 2, collagen I, osteocalcin, and osteonectin. In conclusion, BIO upregulated GSK3β phosphorylation and inhibited its activity, thereby activating the Wnt/β-catenin signaling pathway and promoting the osteogenic differentiation of canine BMSCs. The effect of 1.0 μM BIO on BMSCs differentiation was stronger than that of 0.5 μM BIO.

Transplantation of Bone Marrow-Derived Mesenchymal Stem Cells, Platelet-Rich Plasma, and Fibrin Glue for Periodontal Regeneration.

Appropriate regeneration of periodontal tissues is the primary purpose of periodontal disease treatment. The present study assessed the impact of three key regenerative elements-platelet-rich plasma (PRP), canine bone marrow mesenchymal stem cells (cBM-MSCs), and fibrin glue-on periodontal regeneration. In each of the study's five dogs, Class II furcation defects were established on the buccal surface of five teeth. One tooth (five total) from each dog was placed into one of five groups: (1) PRP + fibrin glue, (2) PRP + fibrin glue + cBM-MSCs, (3) fibrin glue, (4) cBM-MSCs + fibrin glue, and (5) control (no treatment). Histologic and histometric evaluations were performed to assess the formation of new bone, cementum, and the periodontal ligament. Different types of new bone and cementum, the maximum thickness of new cementum and the periodontal ligament (PDL), the vitality of bone, and the presence of inflammation or foreign-body reactions were also elucidated. The histologic and histometric evaluations revealed substantial differences in all groups between the observed maximum thickness of newly formed cementum and PDL. The percentage of bone and cement formation drastically increased with the combined presence of stem cells, fibrin glue, and PRP. The results showed that the inherent regenerative capacity of periodontal tissues can be sufficient if their latent self-repair mechanisms are stimulated.

Analytic characterization of flow cytometric assays for detection of immunoglobulin G on canine erythroid cells, including detection of dog erythrocyte antigen 1 on erythroid precursors.

OBJECTIVE To develop and characterize flow cytometric assays for detecting IgG bound to canine erythrocytes and bone marrow erythroid precursors. SAMPLE Blood samples from 20 healthy and 61 sick dogs with (n = 33) or without (28) immune-mediated hemolytic anemia (IMHA) and bone marrow samples from 14 healthy dogs. PROCEDURES A flow cytometric assay for measurement of IgG on RBCs was developed, and appropriate positive control cells were generated. Analytic and diagnostic performance were characterized. The RBC IgG assay was then combined with density-gradient fractionation of aspirated bone marrow cells and a 2-color process to yield an assay for detecting IgG on nucleated RBCs (nRBCs). Cell sorting and cytologic examination confirmed target cell populations, and anti-dog erythrocyte antigen 1 (DEA1) blood-typing serum was used to generate IgG-positive nRBCs. RESULTS Within- and between-run coefficients of variation for the RBC IgG assay were 0.1% to 13.9%, and > 90% of spiked IgG-positive RBCs were detected. Diagnostic sensitivity and specificity of the assay for detection of IMHA were 88% and 93%, respectively. Cytologic findings for sorted bone marrow fractions rich in early-, mid-, and late-stage nRBCs from 3 healthy dogs indicated 89% to 98% nRBC purity. After IgG coating with anti-DEA1 blood-typing serum, IgG was detected on nRBCs from DEA1-positive, but not DEA1-negative, healthy dogs. CONCLUSIONS AND CLINICAL RELEVANCE The developed RBC IgG assay had favorable analytic and diagnostic performance for detection of IMHA in dogs and was successfully adapted to detect IgG on canine nRBCs of various maturation stages. The findings supported the presence of DEA1 on canine nRBCs.

Research of dental implant inserted in 3‐dimensional hydroxyapatite scaffolds with stripe patterns on pore surface for repairing canine segmental mandibular bone defects

Research of dental implant inserted in 3‐dimensional hydroxyapatite scaffolds with stripe patterns on pore surface for repairing canine segmental mandibular bone defects

Biocompatibility of Titanium Implants with Porous Surface Fabricated by Micromachining

Titanium implants with porous surface arranged orderly were fabricated by micromachining through photolithography and chemical etching. The titanium discs were implanted into the canine mandible alveolar bone for 3 and 21 days, for animal experiments. Neogenetic bone was observed onto porous surface after 7 days‘ implantation. This result allows us to expect application of titanium with porous surface as biomaterials.

An allogenic therapeutic strategy for canine spinal cord injury using mesenchymal stem cells.

This study was conducted to characterize canine bone marrow-derived mesenchymal stem cells (BMSCs); in vivo tracking in mice, and therapeutic evaluation in canine clinical paraplegia cases. Canine BMSCs were isolated, cultured, and characterized in vitro as per International Society for Cellular Therapy criteria, and successfully differentiated to chondrogenic, osteogenic, and adipogenic lineages. To demonstrate the homing property, the pGL4.51 vector that contained luciferase reporter gene was used to transfect BMSCs. Successfully transfected cells were injected around the skin wound in mice and in vivo imaging was done at 6, 12 and 24 hr post MSCs delivery. In vivo imaging revealed that transfected BMSCs migrated and concentrated predominantly toward the center of the wound. BMSCs were further evaluated for allogenic therapeutic potential in 44 clinical cases of spinal cord injuries (SCI) and compared with conventional therapy (control). Therapeutic potential as evaluated by different body reflexes and recovery score depicted significantly better results in stem cell-treated group compared to control group. In conclusion, allogenic canine BMSCs can serve as potent therapeutic candidate in cell-based therapies, especially for diseases like SCI, where the conventional medication is not so promising.