Platelets Rich Plasma Increases Antioxidant Defenses of Tenocytes via Nrf2 Signal Pathway.

Tendinopathies are common disabling conditions in equine and human athletes. The etiology is still unclear, although reactive oxygen species (ROS) and oxidative stress (OS) seem to play a crucial role. In addition, OS has been implicated in the failure of tendon lesion repair. Platelet-rich plasma (PRP) is rich in growth factors that promote tissue regeneration. This is a promising therapeutic approach in tendon injury. Moreover, growing evidence has been attributed to PRP antioxidant effects that can sustain tissue healing. In this study, the potential antioxidant effects of PRP in tenocytes exposed to oxidative stress were investigated. The results demonstrated that PRP reduces protein and lipid oxidative damage and protects tenocytes from OS-induced cell death. The results also showed that PRP was able to increase nuclear levels of redox-dependent transcription factor Nrf2 and to induce some antioxidant/phase II detoxifying enzymes (superoxide dismutase 2, catalase, heme oxygenase 1, NAD(P)H oxidoreductase quinone-1, glutamate cysteine ligase catalytic subunit and glutathione, S-transferase). Moreover, PRP also increased the enzymatic activity of catalase and glutathione S-transferase. In conclusion, this study suggests that PRP could activate various cellular signaling pathways, including the Nrf2 pathway, for the restoration of tenocyte homeostasis and to promote tendon regeneration and repair following tendon injuries.

Animal models of Soft Tissue Sarcoma for alternative anticancer therapy studies: characterization of the A-72 Canine Cell Line.

Canine Soft Tissue Sarcoma (STS) cell line A-72 has been largely employed for antiviral and antiproliferative studies. However, there are few information on their characteristics. Our aim was to evaluate A-72 expression level of genes and proteins involved in the innate immune response and cell cycle, their ability to respond to infective stressors and their possible use as a cellular model for anti-cancer studies in human and animal medicine. For this purpose, we evaluated the basal expression of immune-related, cell cycle and DNA repair genes on this cell line and tumoral tissues. A-72 ability to respond to a wild-type strain of Salmonella typhimurium was assessed. S. typhimurium showed ability to penetrate A-72 causing pro-inflammatory response accompanied by a decrease of cell viability. IL10 and IL18 genes were not expressed in A-72 while CXCL8, NOS2, CXCR4 and PTEN were highly expressed in all samples and TP53 was slightly expressed, as shown in human STS. Our results outline the ability of A-72 to respond to a bacterial agent by modifying the expression of important genes involved in innate immune response and provide a useful model for in vitro evaluation of new therapeutic approaches that could be translated into the human oncology.

Radiographic findings and anatomical variations of the caudal cervical area in horses with neck pain and ataxia: case-control study on 116 horses.

BACKGROUND: Abnormalities of the ventral lamina of the sixth cervical vertebra (AVL-C6) are thought to exert abnormal stress on the articular process joints (APJs) of the cervicothoracic junction. The aim of the study was to investigate the association between AVL-C6 and radiographic findings in the caudal cervical area and between clinical signs of neck pain and ataxia and radiographic findings. METHODS: Medical records of horses subjected to cervical radiography were reviewed. Horses were classified into those with neck pain (group C), those with ataxia (group A) and healthy horses (group H). Presence of AVL-C6 and increased size, dysplasia, remodelling, fragmentation and osteochondral fragment at the APJs (C5-T1) were recorded. Univariable logistic regression analysis was performed to identify the associations between explanatory and dependent variables. Variables with P<0.2 were included in the multivariable analysis. RESULTS: One hundred and sixteen horses were included (44 in group C, 29 in group A, 66 in group H); 24 of 116 horses had radiographic AVL-C6. Age, AVL-C6 and overall/C6-C7 increase in size remained in the final models. CONCLUSIONS: The presence of AVL-C6 and moderate/severe increase in size of the caudal cervical APJs increase the odds of showing neck pain and, if severe, ataxia.

3D Biomimetic Porous Titanium (Ti6Al4V ELI) Scaffolds for Large Bone Critical Defect Reconstruction: An Experimental Study in Sheep.

The main goal in the treatment of large bone defects is to guarantee a rapid loading of the affected limb. In this paper, the authors proposed a new reconstructive technique that proved to be suitable to reach this purpose through the use of a custom-made biomimetic porous titanium scaffold. An in vivo study was undertaken where a complete critical defect was experimentally created in the diaphysis of the right tibia of twelve sheep and replaced with a five-centimeter porous scaffold of electron beam melting (EBM)-sintered titanium alloy (EBM group n = 6) or a porous hydroxyapatite scaffold (CONTROL group, n = 6). After surgery, the sheep were allowed to move freely in the barns. The outcome was monitored for up to 12 months by periodical X-ray and clinical examination. All animals in the CONTROL group were euthanized for humane reasons within the first month after surgery due to the onset of plate bending due to mechanical overload. Nine months after surgery, X-ray imaging showed the complete integration of the titanium implant in the tibia diaphysis and remodeling of the periosteal callus, with a well-defined cortical bone. At 12 months, sheep were euthanized, and the tibia were harvested and subjected to histological analysis. This showed bone tissue formations with bone trabeculae bridging titanium trabeculae, evidencing an optimal tissue-metal interaction. Our results show that EBM-sintered titanium devices, if used to repair critical bone defects in a large animal model, can guarantee immediate body weight-bearing, a rapid functional recovery, and a good osseointegration. The porous hydroxyapatite scaffolds proved to be not suitable in this model of large bone defect due to their known poor mechanical properties.

Proteome Alterations in Equine Osteochondrotic Chondrocytes.

Osteochondrosis is a failure of the endochondral ossification that affects developing joints in humans and several animal species. It is a localized idiopathic joint disorder characterized by focal chondronecrosis and growing cartilage retention, which can lead to the formation of fissures, subchondral bone cysts, or intra-articular fragments. Osteochondrosis is a complex multifactorial disease associated with extracellular matrix alterations and failure in chondrocyte differentiation, mainly due to genetic, biochemical, and nutritional factors, as well as traumas. This study describes the main proteomic alterations occurring in chondrocytes isolated from osteochondrotic cartilage fragments. A comparative analysis performed on equine osteochondrotic and healthy chondrocytes showed 26 protein species as differentially represented. In particular, quantitative changes in the extracellular matrix, cytoskeletal and chaperone proteins, and in cell adhesion and signaling molecules were observed in osteochondrotic cells, compared to healthy controls. Functional group analysis annotated most of these proteins in "growth plate and cartilage development", while others were included in "glycolysis and gluconeogenesis", "positive regulation of protein import", "cell-cell adhesion mediator activity", and "mitochondrion nucleoid". These results may help to clarify some chondrocyte functional alterations that may play a significant role in determining the onset and progression of equine osteochondrosis and, being related, of human juvenile osteochondrosis.

Protective effects of platelet-rich plasma against lidocaine cytotoxicity on canine articular chondrocytes.

BACKGROUND: Lidocaine (LD) is one of the most commonly used local anesthetics for performing arthroscopic surgery and managing of osteoarthritic pain in both human and veterinary medicine. However, over the last years, several studies have focused on the chondrotoxic effects of LD. In order to ensure that intra-articular lidocaine is safe to use, treatments aimed at mitigating chondrocyte death have recently been investigated. The aim of this study is to evaluate the possible protective effects of platelet-rich plasma (PRP) against LD cytotoxicity on canine articular chondrocytes. RESULTS: Articular canine chondrocytes, were exposed to 1% or 1.8% LD alone or in co-presence with 10% PRP for 30 min. In order to evaluate the effects of PRP pre-treatments, experiments were carried out on cells cultured in serum-free medium-or in medium supplemented with 10% PRP or 10% fetal bovine serum. Cell viability was evaluated by methyl thiazolyl tetrazolium assay and cell apoptosis was analyzed by flow cytometry using annexin V-fluorescein isothiocyanate/propidium iodide. The results showed that LD significantly reduced canine chondrocytes viability, probably due to apoptosis induction. Pre-treatment or the co-presence of PRP in the media restored the number of viable chondrocytes. The PRP also seemed to protect the cells from LD-induced apoptosis. CONCLUSIONS: Pre-treatments and/or the simultaneous administration of PRP reduced LD-induced cytotoxicity in canine chondrocytes. Further in vivo studies are required to determine whether PRP can be used as a save protective treatment for dogs receiving intra-articular LD injections.

Adult-onset cyclic esotropia: a case report.

PURPOSE: This case report presents the diagnosis and management of an adult patient with cyclic esotropia, a rare and poorly understood form of strabismus, that is characterized by alternating periods of manifest strabismus and orthotropia. Few cases of adult-onset cyclic strabismus have been reported. The etiology, pathogenesis, and treatment for this condition are reviewed. CASE REPORT: A 37-year-old man with high unilateral myopia and anisometropic amblyopia in his left eye developed cyclic esotropia 2 months after a scleral buckle procedure for a retinal detachment. A 48-hour cycle that consisted of a 24-hour period of orthotropia followed by a 24-hour period of constant left esotropia was present. Magnetic resonance imaging, electromyography, electroencephalography, and ocular and orbital echography were unremarkable, as was a neurological assessment that included fatigue and edrophonium testing. The patient was successfully managed with muscle surgery. CONCLUSIONS: The etiology and pathogenesis of adult-onset cyclic esotropia remain unknown. Cyclic esotropia should be considered in the differential diagnosis when an individual presents with a history of intermittent strabismus that is present some days and not others.