Ex vivo studies involve the examination of biological processes outside of a living organism, typically in a controlled laboratory environment. In equine research, ex vivo studies are used to investigate various physiological and pathological processes in tissues, cells, or organs derived from horses. These studies provide insights into cellular mechanisms, tissue responses, and the effects of various treatments or conditions without the complexities of a whole living system. Common applications in equine research include studying cartilage degradation, tendon repair, and immune responses. This page compiles peer-reviewed research studies and scholarly articles that explore the methodologies, findings, and implications of ex vivo studies in equine science.
Hammond SA, Horohov D, Montelaro RC.Naive T cells can be activated both in vivo and in vitro by specialized antigen presenting cells, dendritic cells (DC), with potent antigen-specific, immunostimulatory activity. Indeed, DC can provide an extremely powerful and important immunological tool by which to potentiate the immune response for specific recognition of foreign antigens. Until recently, the direct isolation of DC from PBMC required laborious procedures with extremely poor yields (<0.1%). Methods have been developed for the human, lower primate, and murine model systems to propagate large numbers of DC from PBMC or bone...
Lees P, May SA, Hoeijmakers M, Coert A, Rens PV.The pharmacodynamics and enantioselective pharmacokinetics of vedaprofen were studied in six ponies in a two period cross-over study, in which a mild acute inflammatory reaction was induced by carrageenan soaked sponges implanted subcutaneously in the neck. Vedaprofen, administered intravenously at a dosage of 1 mg/kg, produced significant and prolonged inhibition of ex vivo serum thromboxane B2 (TXB2) synthesis and short-lived inhibition of exudate prostaglandin E2 (PGE2) and TXB2 synthesis. Vedaprofen also partially inhibited oedematous swelling and leucocyte infiltration into exudate. Vedap...
Marr K, Marsh K, Hernandez L, Cunningham FM, Lees P.Leukotrienes, products of the 5-lipoxygenase pathway of arachidonic acid metabolism, possess properties consistent with their involvement in a range of inflammatory diseases. In this study the pharmacokinetics and pharmacodynamics of the selective 5-lipoxygenase inhibitor, fenleuton, have been examined in the horse. Orally administered fenleuton (four 5 mg kg(-1) doses, given once daily) was absorbed from the gastrointestinal tract, and penetrated readily into tissue cage exudate, the ratio of the plasma:exudate AUC0-48h being 0.90+/-0.02 (n=6). Ionophore-stimulated leukotriene (LT) B4 synthes...
Les CM, Stover SM, Taylor KT, Keyak JH, Willits NH.The objective of this study was to examine several simple ex vivo loading conditions for the equine metacarpus, and to evaluate their ability to reproduce the mid-diaphyseal bone surface strain distributions previously reported in vivo at the walk and trot. Distributed axial compressive loads, and 9 different axial compressive point loads at -7.5 kN and -15 kN were applied to metacarpal-distal carpal bone preparations from 6 Thoroughbred horses, aged 1-5 years. The resulting dorsal, medial, palmar, and lateral mid-diaphyseal bone surface axial and shear strains were compared with previously re...
Barton MH, Ferguson D, Davis PJ, Moore JN.Pentoxifylline (7.5 mg/kg) was bolused intravenously to eight healthy horses and was immediately followed by infusion (1.5 mg/kg/h) for 3 h. Clinical parameters were recorded and blood samples were collected for 24 h. Plasma was separated and concentrations of pentoxifylline, its reduced metabolite I, and 6-keto-prostaglandin F1 alpha were determined. Heparinized whole blood was also incubated ex vivo with 1 ng Escherichi coli endotoxin/mL blood for 6 h before determination of plasma tumour necrosis factor activity. The peak plasma concentrations of pentoxifylline and metabolite I occurred at ...
Moore T, Wilcke J, Chilcoat C, Eyre P, Crisman M.Equine neutrophils (PMN) play a critical role in inflammatory processes in horses. The objective of this study was to characterize equine PMN function ex vivo following stimulation with calcium ionophore A23187 (A23187) and phorbol myristate acetate (PMA). These stimulants trigger different branches of the PMN activation process that occurs in vivo. Equine PMN were isolated from the whole blood of six clinically normal geldings using a one-step discontinuous Percoll gradient technique. Neutrophil aggregation, degranulation, and superoxide anion production were evaluated in assay systems which ...
Martin RB, Gibson VA, Stover SM, Gibeling JC, Griffin LV.Fatigue or stress fractures are an important clinical problem in humans as well as racehorses. An important question in this context is, when a bone experiences fatigue damage during extreme use, how much is it weakened compared to its original state? Since there are very limited data on this question and stress fractures are common in racehorses, we sought to determine the effect of fatigue loading on the monotonic strength of equine cortical bone. Beams were machined from the dorsal, medial and lateral cortices of the third metacarpal bones of six thoroughbred racehorses. Beams from left and...
Gerber V, Gehr P, Straub R, Frenz M, King M, Im Hof V.The aim of this ex-vivo study on excised tracheas of healthy horses was to characterise the microscopic heterogeneity of mucus quality by a visual grading system based on transparency and to determine whether differences in mucus quality, assessed by a visual grading system, influence tracheal mucus velocity (TMV). Small pieces of each trachea were mounted into a humidified chamber under a microscope. Mucus quality was visually subdivided into four grades (MG) and ciliary beat frequency and TMV were determined. Mucus on excised horse tracheal epithelium does not form a homogenous layer. We obs...
Poitras P, Guay P, Vaillancourt D, Zidane N, Bigras-Poulin M.The main objective of this study was to evaluate two freezing protocols and the effect of agar embedding on survival of day 6.5 equine embryos. A total of 133 embryos were used, in one group (n = 51), embryos were first embedded in agar before the freezing protocol was started. A freezing protocol to -30 degrees C or -33 degrees C was used before plunging embryos into liquid nitrogen (LN2). The embryos were thawed in water at 37 degrees C, evaluated and placed in culture. After 24 h culture, the embryos were evaluated for their morphology and development. No differences were observed between e...
Baxter GM, Moore JN.Different dosages of aspirin were administered (by nasogastric tube) to 3 groups of 5 healthy adult horses to determine the minimal effective dosage needed to decrease serum thromboxane B2 (TxB2) concentrations and to determine the duration of this decrease. When compared with their base-line serum TxB2 concentrations, horses in group 1 (given 5 mg/kg) had a 71% decrease in TxB2 concentrations at 24 hours after aspirin was given and a 86% decrease at 48 hours; serum TxB2 concentrations were back to base-line values by 120 hours. Horses in group 2 (given 10 mg/kg) had a 60% decrease in TxB2 con...
Finco DR, Groves C.Free-flow and stop-flow procedures conducted on 2 female and 2 testosterone-treated castrated male ponies indicated that [14C]inulin and exogenous creatinine clearance values were the same. These results indicated that creatinine was neither reabsorbed nor secreted by the renal tubules and that exogenous creatinine clearance was an accurate method for determining glomerular filtration rate. As in other species which have been studied, endogenous creatinine clearance probably underestimated glomerular filtration rate because of the presence of noncreatinine chromogens in plasma.
Clemmons RM, Lee MR, Bliss EL, Asbury AC, Cook D, Brown V.Superoxide dismutase (SOD), a free radical scavenger with anti-inflammatory activity, was administered IM to horses. Ex vivo platelet aggregation in response to arachidonic acid was monitored to determine whether exogenous SOD altered equine platelet prostaglandin metabolism. Preparations of platelet-rich plasma obtained before SOD administration were incubated with different concentrations of SOD and were aggregated with arachidonic acid. Superoxide dismutase did not exert a demonstrable effect, either ex vivo or in vitro. Aspirin abolished arachidonic acid-induced platelet aggregation in vit...
Eicker SW, Ainsworth DM.A procedure was developed for the collection, preparation, storage, and administration of equine plasma. The technique involved exsanguination of anesthetized donor horses via carotid artery catheterization with a large-bore cannula. Blood was collected into plastic bags, allowed to settle by gravity, then transferred into storage bags and frozen. These were quickly thawed when needed.
Soma LR, Korber K, Anderson T, Hopkins J.The effects of furosemide (0.55 mg/kg IV) on the plasma and urinary fentanyl (PFE UFE) concentrations were studied during steady-state conditions. The PFE during the steady-state period was 0.31 +/- 0.027 ng/ml, with no significant changes occurring, even though the rate of excretion of fentanyl (EX) increased during the 1st hour from 112.0 +/- 21.6 to 534.5 +/- 82.9 ng/minute. The EX returned to control levels within 3 hours, as did the UFE. The injection of furosemide increased glomerular filtration rate from 1.97 +/- 0.21 to 3.81 +/- 0.75 ml/kg/min. The fractional reabsorption decreased fro...
Hinrichs K.Intracytoplasmic sperm injection (ICSI) has recently become efficient enough to be considered for clinical use. With ICSI, one spermatozoa is injected into a mature oocyte. Harvesting of an oocyte ex vivo, followed by ICSI and transfer of the fertilized oocyte to the oviduct, may be applicable when semen quality is insufficient for standard insemination. Sperm injection, followed by in vitro embryo culture to the blastocyst stage, may be used in cases where multiple oocytes are to be fertilized (e.g. when oocytes are collected post-mortem). Nuclear transfer (cloning) of horses is possible but ...
Latorre CAS, de Souza AF, Ramos PAR, Nóbrega FS, van de Graaf GMM, Pereira CAM, Ferrigno CRA, Hendrickson DA, De Zoppa ALDV.The objective of this study was to compare the biomechanical properties in a single cycle axial loading test and the types of failures in two constructs (a 3-hole 4.5-mm dynamic compression plate (DCP) and 7-hole 5.5-mm Y locking compression plate (Y-LCP)) in equine proximal interphalangeal joint (PIJ) arthrodesis. One limb in each pair was randomly assigned to PIJ arthrodesis using a 3-hole 4.5-mm DCP combined with two transarticular 5.5-mm cortical screws, whereas the contralateral limb was submitted to PIJ arthrodesis using a 7-hole Y-shaped 5.0-mm LCP in conjunction with one transarticular...
Sidhu ABS, Rosanowski SM, Davis AM, Griffith JF, Robinson P.To quantitate the iatrogenic injury associated with deep plantar metatarsal fasciotomy performed with Metzenbaum scissors compared with a Y-shaped fasciotome. Methods: Experimental ex vivo surgical study. Methods: Cadaveric hind limbs (n = 20) from 10 sound thoroughbred racehorses. Methods: A plantar metatarsal fasciotomy was performed, extending from the proximal extent of the deep metatarsal fascia, distally. Hind limbs were randomly assigned to 2 groups, undergoing fasciotomy with straight Metzenbaum scissors (n = 10) or a Y-shaped fasciotome (n = 10). Magnetic resonance imaging...
Tulloch PJ, Johnston JD, Barber SM, Gellert CL, Lang HM, Panizzi L.To determine degrees of flexion after arthrodesis of the antebrachiocarpal (ABC) joint, middle carpal (MC), and carpometacarpal (CMC) joints combined (MC/CMC), and carpometacarpal (CMC) joint alone. Methods: Ex vivo study. Methods: Forelimbs (n = 9) from 2- to 10-year-old Quarter Horses (5), Thoroughbred (2), and American Paint Horse (2). Methods: Using 2 locking compression plates, 3 partial carpal arthrodesis techniques were performed. Cables and deadweights were connected to limbs and each angle of flexion determined 3 times using a protractor and then averaged. Control measurements were ob...
Parra N, Jaume M, Boscán K, Hernández A, Mijares A, González M, Alvarado Y, Restrepo J.Trypanosoma equiperdum is the causative agent of dourine, a venereal disease in horses and donkeys. This parasite has a widely distribution, is found in Africa, Asia, Southern and Eastern Europe, Russia, Mexico and Venezuela. The T. equiperdum is morphologically indistinguishable to other Trypanozoon species, however differs from other mammalian trypanosomes due to the fact that it is primarily a tissue parasite, generating cutaneous plaques, swelling of genitalia and neurological signs. The aim of this study was to evaluate the trypanocidal effectiveness of a set of derivatives of thiosemicar...
Seeto WJ, Tian Y, Winter RL, Caldwell FJ, Wooldridge AA, Lipke EA.A common challenge in cell therapy is the inability to routinely maintain survival and localization of injected therapeutic cells. Delivering cells by direct injection increases the flexibility of clinical applications, but may cause low cell viability and retention rates due to the high shear forces in the needle and mechanical wash out. In this study, we encapsulated endothelial colony forming cells (ECFCs) in poly(ethylene glycol)-fibrinogen (PF) hydrogel microspheres using a custom-built microfluidic device; this system supports rapid encapsulation of high cell concentrations (10 million c...
Esposito M, Astolfo A, Cipiccia S, Jones CM, Savvidis S, Ferrara JD, Endrizzi M, Dudhia J, Olivo A.Microscopic imaging of cartilage is a key tool for the study and development of treatments for osteoarthritis. When cellular and sub-cellular resolution is required, histology remains the gold standard approach, albeit limited by the lack of volumetric information as well as by processing artifacts. Cartilage imaging with the sub-cellular resolution has only been demonstrated in the synchrotron environment. Objective: To provide a proof-of-concept demonstration of the capability of a laboratory-based x-ray phase-contrast microscope to resolve sub-cellular features in a cartilage sample. Meth...
Estrada RJ, Alvarado GJ, Vargas A, Vargas J, Vargas D, Chacón R, Razquin P, Vindas R.To determine the proximal diffusion distance of radiopaque contrast medium and mepivacaine/methylene blue solution and incidence of inadvertent intrasynovial and intravascular injections of modified sesamoid nerve block (MASB) when compared with traditional plantar nerve analgesia techniques of the equine distal hind limb. Methods: Ex vivo model: 18 hind limbs; and in vivo model: 5 horses in a crossover study. Methods: In the ex vivo model, a mepivacaine/methylene blue solution was used to compare the diffusion distance between MASB, basisesamoid block (BSB), and traditional low plantar block ...
Jha S, Hall JA, Cherian G, Henry LR, Schlipf JW.Neutrophils are involved in inflammation through leukotriene (LT) production. The predominant proinflammatory leukotriene released from neutrophils is LTB4, which serves as a biological marker of inflammation. The purpose of this study was to optimize the conditions ex vivo for LTB4 production by neutrophils from horses and dogs, and platelets from chickens. Optimal production of LTB4 was characterized by incubation time (2.5, 5, 10, 15 or 20 min), temperature (25 or 37 degrees C), and calcium ionophore A23187 concentration (0.1, 1, 10 or 20 microM). Incubation longer than 2.5 min did not incr...
Viljanto M, Kaabia Z, Taylor P, Hincks P, Muir T, Habershon-Butcher J, Bailly-Chouriberry L, Scarth J.Boldenone is an anabolic-androgenic steroid (AAS) that is prohibited in equine sports. However, in certain situations, it is endogenous, potentially formed by the microbes in urine. An approach to the differentiation based on the detection of the biomarkers Δ1-progesterone, 20(S)-hydroxy-Δ1-progesterone and 20(S)-hydroxyprogesterone was assessed, and their concentrations were monitored in the urine of untreated female horses (n = 291) alongside boldenone, boldienone, testosterone and androstenedione. Using an ultra-sensitive analytical method, boldenone (256 ± 236 pg/mL, n = 2...
Keller SA, Valet S, Martens A, Weisse B, Fürst AE, Kümmerle JM. The aim of this study was to evaluate cyclic fatigue behaviour of a new pin with a thread run-out design in comparison with three other types of pins commonly used for equine transfixation pin casting. Methods: Twenty-four pairs of equine cadaveric third metacarpal bones (MC3) equipped with one transfixation pin placed horizontally in the distal metaphysis were tested using a simplified model, mimicking the biomechanical situation of equine transfixation pin casting. A 6.3/8.0-mm Imex Duraface pin with thread run-out design (ITROP) was compared with a 6.1-mm smooth Steinmann pin (SSP), a...
Peek SF, Borah S, Semrad S, McGuirk S, Slack JA, Patton E, Coombs D, Lien L, Darien BJ.Accurate determination of plasma endotoxin concentration is critical for ex vivo and in vitro cellular and molecular studies of endotoxemia in horses. However, reports are conflicting with respect to anticoagulant, handling, and sample preparation. Objective: The purpose of this study was to determine the effect of blood sample fraction and handling time on measurement of endotoxin concentration in horses. Methods: Whole blood, anticoagulated with 3.8% (0.12 M) sodium citrate (9:1), was collected from 5 healthy horses. Whole blood (WB), platelet-rich plasma (PRP), and platelet-poor plasma (PPP...
Mills EP, Liu CC, Emelogu U, Carter RT, Camacho-Luna P, Lewin AC.The purpose of this study was to validate the use of the Reichert Tono-Vera® Vet tonometer rabbit setting in normal ex vivo rabbit eyes and to compare the rabbit setting to the dog, cat, and horse settings of this tonometer. Methods: Six freshly enucleated normal rabbit eyes were cannulated and connected to a fluid reservoir and physiologic monitor. Triplicate measurements were obtained with the four available settings: dog, cat, horse, and rabbit at various intraocular pressures (IOP) ranging from 5 to 80 mmHg. Bland-Altman analysis was utilized to determine bias and 95% limits of agreemen...
Lalanne C, Bonilla AG.To evaluate the feasibility and limitations associated with a minimally invasive ultrasound-assisted cutting thread technique for tenotomy of the deep digital flexor tendon (DDFT) in horses. Methods: Ex vivo study. Methods: Twenty cadaveric forelimbs. Methods: Forelimbs were placed on a jig to mimic a standing semiflexed position and the midmetacarpal region was prepared to perform tenotomy of the DDFT using a percutaneous technique with a cutting thread. For that purpose, the thread was placed percutaneously around the DDFT (first dorsally and then palmarly) with the aid of a curved 20 gauge ...
De Gasperi D, El Azzi MS, Martins JPN, Brounts SH.To describe the feasibility of a novel thread-transecting technique for the tenotomy of the equine deep digital flexor tendon (DDFT). Methods: 39 equine distal limb specimens. Methods: Under ultrasonographic guidance, a surgical thread was percutaneously placed around the DDFT through 2 needle punctures (lateral and medial) using a Tuohy needle in equine limbs (22 forelimbs, 17 hindlimbs). The DDFT was transected by a back-and-forth motion of the thread until the loop emerged from the entry puncture site. Each specimen was dissected and assessed for completeness of transection and iatrogenic d...
