Use of Knotless Barbed Sutures in Laparoscopic Inguinal Hernioplasty in Horses: 40 Cases.

Inguinal hernias (IHs) and ruptures are a relatively common condition in horses, occurring in foals (congenital) and adult (acquired) animals. A retrospective observational analysis was conducted on 40 cases that underwent laparoscopic surgery to close the VRs using barbed sutures alone or combined with other techniques. Signalment, clinical presentation, surgery, and follow-up data were obtained. In total, fifty-nine VRs were closed using barbed sutures (alone or in combination with other methods), with six cases performed prophylactically and forty-four due to acquired IH. Of the forty-four cases with IH, four were non-strangulated hernias, while thirty presented with strangulated small intestines (twenty-eight acquired and two congenital). The results obtained in this study suggest that laparoscopic hernioplasty with barbed sutures is an effective and safe surgical procedure that could be recommended as a standard practice for managing inguinal hernias in horses, particularly when sparing testicles or preserving reproductive capabilities is a priority.

Simple Tube Centrifugation Method for Platelet-Rich Plasma (PRP) Preparation in Catalonian Donkeys as a Treatment of Endometritis-Endometrosis.

The aim of this study was to standardize a simple, manual platelet-rich plasma (PRP) protocol in Catalonian donkeys using single-spin tube centrifugation as a treatment for jenny endometritis. The objective was to obtain a blood product with a moderate concentration of platelets (2 or 3 times baseline physiologic values) and a low WBC (White Blood Cells) concentration. Blood was drawn from six Catalonian donkeys using acid citrate dextrose (ACD) as an anticoagulant, and then processed by single centrifugation at 133× g for two different centrifugation times (10 and 15 min). The PRP samples were evaluated by flow cytometry, and TGF-ß1 (Transforming Growth Factor-Beta1) concentrations were determined by enzyme-linked immunosorbent assay (ELISA). The 10 min centrifugation protocol resulted in a slightly greater release of TGF-ß1 (6044.79 ng/mL), a 2.06-fold increase in platelet concentration, and a 15-fold reduction in leukocyte concentration when compared to the initial values. The 15 min centrifugation time resulted in a 2.44-fold increase in baseline platelet concentration, a reduction in WBC count by a factor of 20, and slightly lower TGF levels (5206 ng/mL). We conclude that both protocols are adequate for the obtention of PRP, and both may have an acceptable therapeutic potential for use in this species, although this needs to be further validated.

Accelerometric Changes before and after Capacitive Resistive Electric Transfer Therapy in Horses with Thoracolumbar Pain Compared to a SHAM Procedure.

Capacitive resistive electric transfer (CRET), a radiofrequency at 448 kHz, increases flexibility in quadricep muscles of human athletes. To assess whether CRET would result in clinical and biomechanical improvements in horses with thoracolumbar pain, 18 sport horses were divided into two groups: CRET (n = 9), subjected to four CRET sessions, during two consecutive weeks, and SHAM (n = 9), subjected to the same procedure with the device off. Clinical examination and accelerometry were performed before and after the four sessions. During the study, horses were in training and in active competition, and did not receive any other treatment. Mann-Whitney and a Wilcoxon matched pair tests were used to compare between the SHAM and CRET groups and before and after the intervention, respectively. CRET horses showed increased dorsoventral (p < 0.002), mediolateral and total power (p < 0.01) after the intervention, suggesting increased back flexibility. SHAM horses did not show any of these modifications after the intervention. No changes were found in the dorsoventral displacement of the gravity center in either group. Thoracolumbar pain decreased one degree after CRET (p = 0.002), and it did not change after SHAM. Epaxial muscle pain decreased two degrees after CRET (p = 0.03) and one degree after SHAM (p = 0.01). These results reflected that CRET therapy would increase back flexibility and decrease thoracolumbar and epaxial pain.

Small Intestine Ultrasound Findings on Horses Following Exploratory Laparotomy, Can We Predict Postoperative Reflux?

Postoperative reflux (POR) is a well-recognized complication after colic surgery in horses, particularly when presenting small intestinal pathology. Even though much has been written about the pathophysiology and management of POR, additional clinical studies are needed to better understand and anticipate this complication. The aim of the study was to provide clinical evidence of ultrasound findings in the postoperative period (three days). The study is based on transcutaneous abdominal ultrasounds of the caudoventral abdomen during the postoperative period (three days), in 58 horses, presented for an exploratory laparotomy, and compared to 20 horses that underwent general anesthesia for an elective surgical procedure. Small intestine (SI) images and videos were analyzed for loop number, loop diameter, wall thickness, motility, and echogenic type of loop contents. Ultrasound findings of horses that had a large colon pathology were similar to those of the control group. Interestingly, horses that presented an SI pathology had significantly thicker SI walls, increased loop diameter, slower motility, and hypoechoic contents, particularly in horses that had undergone small intestinal resection and anastomosis. Although the number of horses that developed POR in our study was too small for statistical analysis, they all had the aforementioned ultrasonographic changes. Abdominal ultrasound, during the postoperative period (three days), was a useful method to identify horses with abnormal small intestinal parameters. Further investigation as to whether these parameters can be used to predict POR in a larger population is warranted.

Changes on the Structural Architecture and Growth Factor Release, and Degradation in Equine Platelet-Rich Fibrin Clots Cultured Over Time.

The aims of the present study were (1) to describe the microscopic and ultrastructural appearance of equine platelet-rich fibrin (PRF) clots and (2) to determine the release and degradation of transforming growth factor beta 1 (TGF-ß1) and insulin-like growth factor type I (IGF-I) from PRF clots incubated over 14 days. Whole blood from six horses was collected into plain tubes and centrifuged at 240 g for 8 minutes. Clots were evaluated by histology and by both transmission and scanning electronic microscopy (TEM and SEM). Growth factor concentrations were measured by ELISA at 48-hour intervals over 14 days and analyzed by one-way repeated-measures ANOVA. Histology showed a clot composed by a fibrin layer and a cellular layer with platelets and leukocytes. Scanning electron microscopy showed the cells trapped by an incipient fibrin network at 1 hour. At day 8, these cells were embedded by an incipient fibrin network. At day 14, the leukocytes and platelet aggregates from the clot were imbibed in an organized web of fibrin fibrils. TEM exhibited platelets with preserved cytoplasm and alpha granules randomly scattered at day 8, and damaged platelets with interrupted cytoplasm and organelle emigration to the periphery at day 14. TGF-ß1 and IGF-I concentrations showed a progressive increase until day 14. TGF-ß1 was released from PRF clots in a gradual and controlled manner, and increasing its concentration for two weeks, which supports TEM findings indicating that platelets began disintegrating by day 14. Furthermore, IGF-I production and release from PRF clots is sustained over time.

Anatomical variations of the equine popliteal tendon.

The function of the popliteal muscle and tendon in horses remains undescribed. In humans, it is considered a stabilizer of the posterior-lateral region of the knee; its function is closely related to that of the lateral collateral ligament (LCL) and meniscus. The popliteal tendon (PopT) constitutes the main proximal attachment of the popliteus muscle to the femur, and in humans, insertional variations have been described. Knowledge of anatomical variations is needed for the correct interpretation of diagnostic images and arthroscopic findings. To elucidate further the anatomy of the equine PopT, both hind limbs of 30 horses were dissected. Similar to humans, the equine PopT has 3 variants (types I, II, and III) depending on the number of components forming the tendon. Additionally, the area of insertion varies; the location can be either cranial, underneath, or caudal to the proximal insertion of the LCL. Furthermore, the PopT has a constant attachment to the lateral meniscus. The results of the present study are useful for clinicians working with equine orthopedics, as the tendon and insertional variants could affect the interpretation of diagnostic images and arthroscopic examinations.

Assessment of effectiveness and safety of repeat administration of proinflammatory primed allogeneic mesenchymal stem cells in an equine model of chemically induced osteoarthritis.

BACKGROUND: This study aimed at assessing the effectiveness and safety of repeated administrations of allogeneic bone marrow-derived mesenchymal stem cells (BM-MSCs) primed with tumor necrosis factor (TNF)-α and interferon-γ in an equine model of chemically-induced osteoarthritis. Arthritis was induced in both radio-carpal (RC)-joints by amphotericin-B in 18 ponies, divided into three groups depending on the treatment injected: MSC-naïve (n = 7), MSC-primed (n = 7) and control (n = 4). The study consisted of two phases and used one RC-joint of each animal in each phase, with four months time-lapse, in order to assess two end-points. Clinical, synovial, radiological and ultrasonographic follow-up was performed. At six months, animals were euthanized and both carpi were assessed by magnetic resonance imaging (MRI), gross anatomy, histopathology, histochemistry and gene expression. RESULTS: Clinical and synovial inflammatory signs were quicker reduced in MSC-treated groups and repeated allogeneic administration did not produce adverse reactions, but MSC-primed group showed slight and transient local inflammation after second injection. Radiology and MRI did not show significant differences between treated and control groups, whereas ultrasonography suggested reduced synovial effusion in MSC-treated groups. Both MSC-treated groups showed enhanced cartilage gross appearance at two compared to six months (MSC-naïve, p < 0.05). Cartilage histopathology did not reveal differences but histochemistry suggested delayed progression of proteoglycan loss in MSC-treated groups. Synovium histopathology indicated decreased inflammation (p < 0.01) in MSC-primed and MSC-naïve at two and six months, respectively. At two months, cartilage from MSC-primed group significantly (p < 0.05) upregulated collagen type II (COL2A1) and transforming growth factor (TGF)-ß1 and downregulated cyclooxygenase-2 and interleukin (IL)-1ß. At six months, MSC-treatments significantly downregulated TNFα (p < 0.05), plus MSC-primed upregulated (p < 0.05) COL2A1, aggrecan, cartilage oligomeric protein, tissue inhibitor of metalloproteinases-2 and TGF-ß1. In synovium, both MSC-treatments decreased (p < 0.01) matrix metalloproteinase-13 expression at two months and MSC-primed also downregulated TNFα (p < 0.05) and IL-1ß (p < 0.01). CONCLUSIONS: Both MSC-treatments provided beneficial effects, mostly observed at short-term. Despite no huge differences between MSC-treatments, the findings suggested enhanced anti-inflammatory and regulatory potential of MSC-primed. While further research is needed to better understand these effects and clarify immunogenicity implications, these findings contribute to enlarge the knowledge about MSC therapeutics and how they could be influenced.

Long-term cytokine and growth factor release from equine platelet-rich fibrin clots obtained with two different centrifugation protocols.

OBJECTIVES: To compare the temporal release (over three weeks) of tumor necrosis factor alpha (TNF-α), interleukin 4 (IL-4), IL-1 receptor antagonist (IL-1ra), platelet-derived growth factor BB (PDGF-BB) and transforming growth factor beta-1 (TGF-ß1) from two platelet-rich fibrin (PRF) preparations from equine blood obtained at either 240g/8min or 416g/10min. METHODS: Whole blood from 10 horses was used to obtain PRF clots by two different centrifugation protocols. After 1h of rest, PRF clots were deposited in wells with culture medium, which was changed at 6h, 24h and then every 48h to 21days. Cytokines and GFs were measured by ELISA at 1h (serum supernatants from PRF clots) and all time points of culture medium change. A negative control (plasma) and a positive control (blood lysate) were also included. RESULTS: There were no relevant differences between the two protocols for the temporal release of proteins. However, a significant (p=0.01) effect of time was noted. All cytokines were detected after 6h of PRF clot culture until day 21. GF were detected at 1h until day 21. The concentrations for these proteins diminished gradually over time. A highly significant (p=0.01) correlation was noticed between all the proteins evaluated. CONCLUSIONS: Leukocytes enmeshed in PRF clots were able to produce cytokines, TGF-ß1 and PDGF-BB. These findings demonstrate a paramount role of leukocytes in wound healing induced or modified by PRF clots in mammals.

Inflammation affects the viability and plasticity of equine mesenchymal stem cells: possible implications in intra-articular treatments.

Mesenchymal stem cells (MSCs) are gaining relevance for treating equine joint injuries because of their ability to limit inflammation and stimulate regeneration. Because inflammation activates MSC immunoregulatory function, proinflammatory priming could improve MSC efficacy. However, inflammatory molecules present in synovial fluid or added to the culture medium might have deleterious effects on MSCs. Therefore, this study was conducted to investigate the effects of inflammatory synovial fluid and proinflammatory cytokines priming on viability and plasticity of equine MSCs. Equine bone marrow derived MSCs (eBM-MSCs) from three animals were cultured for 72 h in media supplemented with: 20% inflammatory synovial fluid (SF); 50 ng/mL IFN-γ and TNF-α (CK50); and 20 ng/mL IFN-γ and TNF-α (CK20). Proliferation assay and expression of proliferation and apoptosis-related genes showed that SF exposed-eBM-MSCs maintained their viability, whereas the viability of CK primed-eBM-MSCs was significantly impaired. Tri-lineage differentiation assay revealed that exposure to inflammatory synovial fluid did not alter eBM-MSCs differentiation potential; however, eBM-MSCs primed with cytokines did not display osteogenic, adipogenic or chondrogenic phenotype. The inflammatory synovial environment is well tolerated by eBM-MSCs, whereas cytokine priming negatively affects the viability and differentiation abilities of eBM-MSCs, which might limit their in vivo efficacy.

Priming Equine Bone Marrow-Derived Mesenchymal Stem Cells with Proinflammatory Cytokines: Implications in Immunomodulation-Immunogenicity Balance, Cell Viability, and Differentiation Potential.

Mesenchymal stem cells (MSCs) have a great potential for treating equine musculoskeletal injuries. Although their mechanisms of action are not completely known, their immunomodulatory properties appear to be key in their functions. The expression of immunoregulatory molecules by MSCs is regulated by proinflammatory cytokines; so inflammatory priming of MSCs might improve their therapeutic potential. However, inflammatory environment could also increase MSC immunogenicity and decrease MSC viability and differentiation capacity. The aim of this study was to assess the effect of cytokine priming on equine bone marrow-derived MSC (eBM-MSC) immunoregulation, immunogenicity, viability, and differentiation potential, to enhance MSC immunoregulatory properties, without impairing their immune-evasive status, viability, and plasticity. Equine BM-MSCs (n = 4) were exposed to 5 ng/mL of TNFα and IFNγ for 12 h (CK5-priming). Subsequently, expression of genes coding for immunomodulatory, immunogenic, and apoptosis-related molecules was analyzed by real-time quantitative polymerase chain reaction. Chromatin integrity and proliferation assays were assessed to evaluate cell viability. Trilineage differentiation was evaluated by specific staining and gene expression. Cells were reseeded in a basal medium for additional 7 days post-CK5 to elucidate if priming-induced changes were maintained along the time. CK5-priming led to an upregulation of immunoregulatory genes IDO, iNOS, IL-6, COX-2, and VCAM-1. MHC-II and CD40 were also upregulated, but no change in other costimulatory molecules was observed. These changes were not maintained 7 days after CK5-priming. Viability and differentiation potential were maintained after CK5-priming. These findings suggest that CK5-priming of eBM-MSCs could improve their in vivo effectiveness without affecting other eBM-MSC properties.

Field-enhanced bulk conductivity and resistive-switching in Ca-doped BiFeO3 ceramics.

The bulk conductivity at room temperature of Ca-doped BiFeO3 ceramics is p-type and increases reversibly by up to 3 orders of magnitude under the influence of a small dc bias voltage in the range ∼3 to 20 V mm(-1). The effect occurs in both grain and grain boundary regions, is isotropic and does not involve creation of filamentary conduction pathways. It is proposed that, by means of capacitive charging and internal ionisation processes under the action of a dc bias, hole creation leads to a more conductive excited state. This gradually returns to the ground state when the dc bias is removed and the holes recombine with electrons trapped at the sample surface. The holes are believed to be created on oxygen, as O(-) ions.

Does intraoperative low arterial partial pressure of oxygen increase the risk of surgical site infection following emergency exploratory laparotomy in horses?

Decreased tissue oxygenation is a critical factor in the development of wound infection as neutrophil mediated oxidative killing is an essential mechanism against surgical pathogens. The objective of this prospective case series was to assess the impact of intraoperative arterial partial pressure of oxygen (PaO2) on surgical site infection (SSI) in horses undergoing emergency exploratory laparotomy for acute gastrointestinal disease. The anaesthetic and antibiotic protocol was standardised. Demographic data, surgical potential risk factors and PaO2, obtained 1h after induction of anaesthesia were recorded. Surgical wounds were assessed daily for infection during hospitalisation and follow up information was obtained after discharge. A total of 84 adult horses were included. SSI developed in 34 (40.4%) horses. Multivariate logistic regression showed that PaO2, anaesthetic time and subcutaneous suture material were predictors of SSI (AUC=0.76, sensitivity=71%, specificity=65%). The use of polyglycolic acid sutures increased the risk and horses with a PaO2 value < 80 mm Hg [10.6 kPa] and anaesthetic time >2h had the highest risk of developing SSI (OR=9.01; 95% CI 2.28-35.64). The results of this study confirm the hypothesis that low intraoperative PaO2 contributes to the development of SSI following colic surgery.

Effects of the breed, sex and age on cellular content and growth factor release from equine pure-platelet rich plasma and pure-platelet rich gel.

BACKGROUND: There is no information on the effects of the breed, gender and age on the cellular content and growth factor (GF) release from equine pure-platelet rich plasma (P-PRP) and pure-platelet rich gel (P-PRG). The objectives of this study were: 1) to compare the cellular composition of P-PRP with whole blood and platelet poor plasma (PPP); 2) to compare the concentration of transforming GF beta 1 (TGF-ß1) and platelet derived GF isoform BB (PDGF-BB) between P-PRP treated with non-ionic detergent (P-PRP+NID), P-PRG (activated with calcium gluconate -CG-), PPP+NID, PPP gel (PPG), and plasma and; 3) to evaluate and to correlate the effect of the breed, gender and age on the cellular and GF concentration for each blood component. Forty adult horses, 20 Argentinean Creole Horses (ACH) and, 20 Colombian Creole Horses (CCH) were included. Data were analyzed by parametric (i.e.: t-test, one way ANOVA) and non parametric (Kruskal-Wallis test, Wilcoxon test) tests. Correlation analysis was also performed by using the Spearman and Pearson tests. A p ≤ 0.05 was set as significant for all tests. All the blood components were compared for platelet (PLT), leukocyte (WBC), TGF-ß1 and PDGF-BB concentrations. The effect of the breed, gender and age on these variables was analyzed. A P ≤ 0.05 was accepted as significant for all the tests. RESULTS: PLT counts were 1.8 and 0.6 times higher in P-PRP than in whole blood and PPP, respectively; WBC counts were 0.5 and 0.1 times lower in P-PRP, in comparison with whole blood and PPP, respectively. TGF-ß1 and PDGF-BB concentrations were 2.3 and 262 times higher, respectively, in P-PRG than in plasma, and 0.59 and 0.48 times higher, respectively, in P-PRG than in PPG. P-PRG derived from CCH females or young horses presented significantly (P < 0.001) higher PDGF-BB concentrations than P-PRG derived from ACH males or older horses. CONCLUSIONS: Our results indicated that P-PRP obtained by a manual method was affected by intrinsic factors such as the breed, gender and age. Equine practitioners should be aware that cellular and GF release from P-PRP/P-PRG could change according with the intrinsic variables associated with a patient in particular.

Synthesis, structural characterization, and electrical properties of new oxygen-deficient tetragonal tungsten bronzes Ba2NdTi(2+x)Nb(3-x)O(15-x/2).

Oxygen-deficient tetragonal tungsten bronzes ceramics with general formula Ba(2)NdTi(2+x)Nb(3-x)O(15-x/2) (0 ≤ x ≤ 1) have been prepared by low temperature solvothermal synthesis with final firing of ceramics at 1100-1300 °C in air. Rietveld refinement of X-ray powder diffraction (XRD) and neutron powder diffraction (ND) data at room temperature of Ba(2)NdTi(3)Nb(2)O(14.5) shows that Ba and Nd are ordered on the 15-coordinate and 12-coordinate sites, respectively, Ti and Nb are disordered nonrandomly over the two octahedral sites, and oxygen vacancies locate preferentially in the coordination sphere of Nd and Ti/Nb(2) atoms. Variable frequency impedance measurements show that samples are poor electronic conductors with activation energies ∼0.8-1.7 eV, conductivities ∼1 × 10(-5) S cm(-1) at ∼725 °C and with some evidence of oxide ion conduction at high x values. Composition dependence of the dielectric properties shows a transition from classic ferroelectric behavior with Ba(2)NdTi(2)Nb(3)O(15) to a relaxor-like behavior with Ba(2)NdTi(3)Nb(2)O(14.5). At intermediate compositions, both a first-order phase transition and relaxor-like behavior are observed.

Determinación de marcadores inflamatorios y anabólicos articulares en caballos jóvenes estabulados con osteocondrosis tarsocrural / Determination of inflammatory and anabolic joint markers in stabled young horses with tarsocrural ostechondrosis

Objetivo. Describir los valores de las concentraciones en líquido sinovial (LS) de prostaglandina E2 (PGE2), óxido nítrico (NO), ácido hialurónico (HA) y proteína total (PT) en caballos jóvenes estabulados con osteocondrosis (OC) tarsocrural y correlacionar los valores de estos marcadores con el recuento total de células del LS, el grado de efusión sinovial (ES) y con el grado de cojera (GC). Materiales y métodos. Las concentraciones en LS de PGE2 fueron determinadas mediante ELISA, las de NO fueron valoradas por la reacción de Greiss, las de HA fueron medidas con radioinmunoanálisis RIA y las de PT mediante refractometría. Resultados. Once caballos entre 8 y 36 meses fueron incluidos. La mediana y el rango (R) de los grados de ES y GC fueron de 1.5 (R: 0.5-2) y 0 (R: 0-2), respectivamente. La mediana de la concentración de PGE2 y NO, respectivamente fue 130.6 pg/mL (R: 41-231.7 pg/mL) y 2.92 mM (R: 1.3-6.3 mM).. La concentración de HA presentó una mediana de 312.4 mg/mL (R: 70-543.5 mg/mL). La concentración promedio de PT fue de 1.2±-0.36 g/dL. Se observó correlación negativa estadísticamente significativa entre las concentraciones sinoviales de HA y el grado de efusión sinovial (ñ=-0.7; p= 0.048) y el grado de cojera (ñ=-0.78; p=0.014). Conclusiones. La concentración de HA en LS de caballos con OC podría estar negativamente correlacionada con la gravedad de los signos clínicos.