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.
Gray AW, Davies ME, Jeffcott LB.Cathepsin K and cathepsin B were immunolocalised in equine osteoclasts (OC s) present in ex vivo cartilage/subchondral bone samples. Samples were obtained post mortem from the lateral trochlear ridge (LTR) of six horses and ponies aged between 303 days gestation to 8 months. Strong expression of cathepsin K was detected in OC s, particularly those located at the osteochondral junction, apparently involved in the resorption of calcified cartilage. Cathepsin K expression was also detected in hypertrophic chondrocytes and in the endothelial cells of some blood vessels penetrating the hypertrophic...
Walter LJ, Davies HM.Accuracy and limitations of a radiographic technique were established for measurement of metacarpal bone shape in horses. A radiographic index (RI) has been used to measure changes in third metacarpal (Mc3) bone shape in response to training in young racehorses. The aim of these experiments was to determine the effects of positioning of the radiographic equipment on RI measurements from lateromedial radiographs of a left ex vivo metacarpus. Repeatability of the RI measurement in left lateromedial and right mediolateral view Mc3s were evaluated. The width of the dorsal cortex (DC), palmar corte...
Parviainen AK, Barton MH, Norton NN.To evaluate effects of polymyxin B sulfate (PMB) on response of horses to endotoxin, using an ex vivo model. Methods: 8 healthy horses. Methods: In a crossover design, 3 doses of PMB (100, 1,000, and 10,000 U/kg of body weight) and physiologic saline solution (control) were evaluated. Prior to and for 24 hours after administration of PMB, blood samples were collected into heparinized tubes for use in 2 assays. For the endotoxin-induced tumor necrosis factor (TNF) assay, blood samples were incubated (37 C for 4 h) with 1 ng of Escherichia coli or Salmonella Typhimurium endotoxin/ml of blood. Pl...
van den Hoogen BM, van den Lest CH, van Weeren PR, van Golde LM, Barneveld A.In this study, the effect of different exercise regimens on proteoglycan metabolism of articular cartilage was examined in 43 newborn foals randomly divided into 3 groups: a) box-rest, b) box-rest with training and c) free pasture exercise. They were subjected to these exercise regimens from ages 1 week to 5 months and at 5 months, 24 foals (8 from each group) were sacrificed to assess short-term exercise effects. The remaining 19 foals were subjected to the same regimen of light exercise for an additional 6 months before being sacrificed to evaluate possible long-term effects. Articular carti...
van den Hoogen BM, van de Lest CH, van Weeren PR, van Golde LM, Barneveld A.In osteochondrosis (OC) the process of endochondral ossification is impaired. Proteoglycans form one of the major components of the extracellular matrix of cartilage and are able to bind calcium. For this reason, proteoglycans are thought to play an important role early in the mineralisation process and may, therefore, be important in the pathogenesis of OC. To investigate possible differences in proteoglycan metabolism, normal and osteochondrotic articular-epiphyseal cartilage was harvested from the hock and stifle joints of 43 foals age 5 and 11 months. The samples were cultured as explants ...
Van Hoogmoed LM, Snyder JR, Nieto JE, Vatistas NJ, Harmon FA.To determine efficacy of an extracorporeal circuit to maintain a segment of equine large colon for 3.5 hours and to evaluate the effect of low arterial flow on histologic and metabolic variables. Methods: Segments of large colon from 15 healthy adult horses. Methods: The pelvic flexure was surgically removed and maintained in an isolated circuit. In the control group, tissue was evaluated for 3.5 hours, whereas in the low-flow group, arterial flow was reduced to 20% of baseline for 40 minutes followed by 2 hours of reperfusion. Various metabolic and hemodynamic variables were evaluated at 30-m...
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...
Verhaar N, Hammer E, Reineking W, Hewicker-Trautwein M, Geburek F.To compare the practicability and tissue sample quality between different intestinal biopsy techniques. Methods: Experimental, randomized ex vivo study. Methods: Small intestine of nine horses. Methods: Four different biopsy techniques were evaluated in the aboral jejunum and the ileum within 1 h after euthanasia. One segment was used as control (C), and the applied techniques included an 8 mm biopsy punch (BP), transverse wedge resection (TW), longitudinal wedge resection with transverse closure (LW) and a longitudinal sample using Eppendorfer biopsy forceps (EF). Defects were closed using ...
Korsós SA, Kibleur P, Josipovic I, Boone M, Vlaminck L.In orthograde endodontic treatments, different methods are available to debride the pulp canals of endodontically compromised equine cheek teeth, but their efficacy is unknown. Objective: To explore and compare the efficacy and anatomical changes caused by manual versus reciprocating filing techniques in equine cheek teeth, to explore the presence of instrumentation mishaps described in human dentistry and to explore anatomical complexities of the pulp cavity that often remain uninstrumented using microcomputed tomography (μCT). Methods: Ex-vivo randomised experiments. Methods: Twenty-two ext...
Nugent Z, Jensen A, Owen N, Peffers AJ, Moothanchery M, Peffers MJ.Equine odontoclastic tooth resorption and hypercementosis (EOTRH) is a painful disorder primarily affecting the incisor teeth of horses over 15 years of age. Clinical signs of the disease include prehension problems, halitosis and in severe cases weight loss. The disease predominately affects the reserve crown and presents as a loss of dental tissue and excessive build-up of cementum. Objective: To determine the radiographic scores of horses with EOTRH and age-matched controls and to increase understanding of EOTRH using microCT to compare teeth from horses with EOTRH and age-matched control...
CT arthrography (CTA) for evaluating cartilage defects would be beneficial in clinical practice. Since the contrast medium (CM) volume is often driven by operator experience, the aim was to evaluate the minimum volume of iodinated CM for CTA sufficient to identify iatrogenic cartilage defects in horse fetlock. The study was conducted on ex-vivo 32 distal limbs from adult horses collected at slaughter. Iatrogenic cartilage defects were arthroscopically made on fetlock, at 5 predetermined locations. Subsequently, CT scans were performed, according to the following protocol: precontrast scan, fol...
Gao T, Liu X, Qiu D, Li Y, Qiu Z, Qi J, Li S, Guo X, Zhang Y, Wang Z, Gao X, Ma Y, Ma T.Cefquinome is used to treat septicemia caused by () and respiratory infections caused by subsp. in foals. However, studies reporting the use of cefquinome to target as pathogens of sepsis are lacking. Therefore, this study aimed to determine the optimal dosage regimen for cefquinome against using a PK/PD model. After the administration of 1 mg/kg cefquinome (intramuscularly or intravenously), blood samples were collected at different time points to determine the serum concentration of cefquinome via HPLC. The pharmacokinetic parameters were evaluated via NCA (WinNonlin 5.2.1 software). Th...
Kodama S, Maeda T, Mita H, Tamura N, Chiba K, Matsukawa M.Despite the prevalence of lower limb disease in racehorses, outdoor bone evaluation equipment is lacking. Consequently, a secure and cost-effective field screening system for bones is required to identify early-stage leg illness. Elastic stiffness cannot be measured using x-ray techniques and is closely associated with bone quality, which is recognized as a significant determinant of fracture risk. Quantitative ultrasound has garnered attention as a viable option, offering insights into elastic stiffness. Understanding the characteristic behaviors of ultrasonic wave properties (velocity, atten...
Kadic LIM, Liu CC, Leise BS, McCauley CT, Riggs LM.The aim of the study was to determine if a novel arthrodesis technique for the equine metacarpophalangeal (MCP) joint using a human non-contact bridging (NCB), polyaxial locking compression plate system has biomechanical and potential clinical advantages versus a traditional 5.5 mm stainless steel locking compression plate (LCP) under biomechanical testing. Methods: An ex vivo biomechanical equine cadaver study. Methods: A total of 11 MCP joints collected from adult Thoroughbred horses euthanized unrelated to musculoskeletal disease. Methods: Each pair of MCP joints were randomly implanted w...
Baldwin CM, Gillen A.To evaluate three different jejunal transection angles for end-to-end jejunojejunostomies, comparing construction time, lumen size, and suture number. Methods: Ex vivo cadaver study. Methods: Eight euthanized horses each had three mid-jejunal specimens harvested. Methods: Jejunal segments were assigned to one of three groups (A30, A45, and A60) and transected at 30, 45, or 60°, respectively. Following transection, a standardized handsewn single-layer interrupted modified Lembert anastomosis was performed, and construction time and number of sutures placed were recorded. Anastomotic index (AI)...
Ysebaert MP, Johnson J, Marie U, Campos A, Verchrerin A, Ducharme NG, Rossignol F, Luedke LK.To compare the biomechanical stability of a novel arytenoid implant (tie-bolt, TB) with a suture anchor (SA) and standard laryngoplasty using suture alone (control) in equine larynges. Methods: Randomized ex vivo controlled experimental study. Methods: Thirty equine cadaver larynges. Methods: Larynges were randomly assigned to TB, SA, or control groups (n = 10 each). The force of abduction was measured for each construct, then constructs underwent cyclic loading (5-50 N for 3000 cycles) to assess elongation, followed by load-to-failure testing to assess ultimate load, displacement, stiffn...
Bosch K, Zsoldos RR, Hartig A, Licka T.The influence of soft tissue structures, including ligaments spanning one or more intervertebral junctions and the nuchal ligament, on motion of the equine cervical joints remains unclear. The present study addressed this using four post-mortem horse specimens extending from head to withers with all ligaments intact. Three-dimensional kinematics was obtained from markers on the head and bone-anchored markers on each cervical and the first thoracic vertebra during rotation, lateral bending, flexion and extension of the whole head, and neck segment. Yaw, pitch, and roll angles in 8 cervical join...
Sullivan J, Blea J, McKemie DS, Kass PH, Knych HK.The pharmacokinetics and pharmacodynamics of betamethasone following intra-articular administration to horses have been described; however, studies characterizing intramuscular administration are lacking. Twenty-four horses received an intramuscular dose of 12 mg betamethasone sodium phosphate/betamethasone acetate. Blood and urine were collected at post administration for up to 408 h. Concentrations of betamethasone were determined using LC-MS/MS and pharmacokinetic parameters determined using a Population PK three-compartment model. The duration of pharmacodynamic effects was assessed by...
Márquez-Moya A, Carreras-Vico N, Sala-Ayala L, Martínez-Boví R, Cuervo-Arango J.Transvaginal ultrasound-guided follicular aspiration or ovum pick-up (OPU) has become the standard technique for oocyte collection in mares for intracytoplasmic sperm injection (ICSI). Although repeated follicular flushes and wall scraping are commonly used to improve oocyte recovery rate (ORR), the relative contribution of each remains unclear. This study aimed to compare the effects of multiple flushes versus controlled scraping time on ORR in mares. A controlled trial was conducted in two phases: (1) an ex vivo phase using slaughterhouse ovaries (n = 32), and (2) an in vivo phase in c...
Aimonetti E, Genton M, Lischer CJ, Rossignol F.To determine the benefit of using a three-dimensional (3D)-printed titanium aiming device for placing two 5.5-mm transarticular screws during proximal interphalangeal joint (PIPJ) arthrodesis in equine species. Methods: Ex vivo study. Methods: A total of 40 cadaveric forelimbs from adult horses. Methods: A senior surgeon and a first-year resident each performed 20 constructs: 10 using the device and 10 using a freehand technique. Procedure duration and number of intraoperative radiographs were recorded. Final radiographs were evaluated by an ECVS-certified surgeon unaware of treatment group, b...
Lepage H, de Chaisemartin C, Spadaro Rosselo A, Leroy H, Lepage O.To describe and report clinical outcomes after transcutaneous guttural pouch (GP) catheterization (TGPC) in standing horses. Methods: Ex vivo study and case series. Methods: One cadaver head, records of 10 normal horses and 14 horses treated with TGPC. Methods: Relevant anatomical landmarks were determined through dissection of one cadaveric specimen and 10 normal radiographic studies. Records of horses diagnosed with empyema or mycosis and treated with standing TGPC were reviewed for complications that occurred during or after the operation. Results: Ex vivo and radiological studies identifie...
Sullivan J, Blea J, Morales CJ, McKemie DS, Kass PH, Knych HK.Corticosteroids, such as isoflupredone, are effective anti-inflammatory medications and as such are commonly used to treat inflammation associated with training and injuries in performance horses. While the pharmacokinetics and pharmacodynamics of isoflupredone acetate (IPA) following intra-articular administration to horses has been well described, studies characterizing intramuscular (IM) administration are lacking. The objective of the current study was to describe the pharmacokinetics and anti-inflammatory effects of IPA following IM administration to horses. Twelve horses received a singl...
Mitlyng N, Hobbs KJ, Cooper BL, Sheats MK.Lipopolysaccharide (LPS) is frequently used in equine research to model clinical endotoxemia; however, there is no standardized protocol for inducing cytokine production in equine whole blood. To address this knowledge gap, the goal of this study was to compare the cytokine responses elicited by three different LPS stimulation protocols. Whole blood was collected from six healthy horses (aged 5-30 years; mixed breeds and genders) from the North Carolina State University teaching herd (IACUC #23-412). Sixty milliliters of heparinized blood were aseptically drawn and divided into 15 mL aliquots....
Beggan CP, Panizzi L, Oliver LJ.To measure microsphere recovery following needle-through-and-through lavage (NTAT) of the metacarpo-/metatarsophalangeal joint (fetlock) and digital flexor tendon sheath (DFTS) compared to endoscopic lavage (EL). Methods: Ex vivo experimental study. Methods: Adult equine cadavers immediately following euthanasia (n = 10). Methods: Colored 15 μm microspheres (2 million) were injected into fetlock joints and DFTS. Synovial structures were assigned to NTAT or EL groups. Each lavage was performed using 5 L of 0.9% NaCl, sequentially collecting egress fluid for microsphere quantification. Re...
Hoerdemann M, Sahoo DK, Allbaugh RA, Kubai MA.To assess if an inexpensive, commercially available ultraviolet C (UV-C) light device with a peak emission of 275 nm can inhibit equine keratomycosis-associated pathogens located at different corneal depths in an ex vivo model. Methods: A controlled, randomized experimental design. Aspergillus fumigatus and Fusarium solani isolates were inoculated in fresh bovine corneoscleral transplants, superficially or at 450 μm of stromal depth for the ulcerative keratomycosis or stromal abscess model, respectively. After a minimum of 18 h of incubation, treatment groups received 15 s of UV-C li...
Profaska M, Zarzycka M, Dubniewicz K, Witkowski M, Wieczorek J, Gil D, Wafula S, Lanh DK, Kotula-Balak M.Mare endometrosis remains a poorly understood pathological process. Objective: Mare endometrial tissue with endometrosis was used to determine the effects of relaxin (RLX). Healthy tissues that were left untreated (H), or treated with vehicle (Hveh), and tissues with endometrosis that were left untreated (E), treated with vehicle (Eveh), and treated with RLX (10, 25, 50 nM; ER) were used for an ex vivo system for 72 h. Methods: Tissue histological examination, and immunoenzymatic measurement of the concentrations of transforming growth factor β (TGF-β), interleukins (IL-6 and IL-8), and pr...
Vishwanath K, McClure SR, Bonassar LJ.Intra-articular injections of hyaluronic acid (HA) are the cornerstone of osteoarthritis (OA) treatments. However, the mechanism of action and efficacy of HA viscosupplementation are debated. As such, there has been recent interest in developing synthetic viscosupplements. Recently, a synthetic 4 wt% polyacrylamide (pAAm) hydrogel was shown to effectively lubricate and bind to the surface of cartilage in vitro. However, its ability to localize to cartilage and alter the tribological properties of the tissue in a live articulating large animal joint is not known. The goal of this study was to...
Buckman PN, Brinker BA, Kapeller LE, Komáromy AM.To determine the accuracy of the EYEMATE-SC suprachoroidal tracer for telemetric tonometry in canine and equine globes. Methods: The EYEMATE-SC sensor (7.8 mm × 3.8 mm × 1 mm) was implanted in the suprachoroidal space of four freshly enucleated normal canine and two normal equine eyes. The anterior chambers were cannulated and connected to a reservoir of Plasma-Lyte A and a manometer. Starting at a manometric IOP of 5 mmHg, the pressure was progressively increased to 80 mmHg by raising the reservoir. At each setpoint (5, 10, 15, 20, 25, 30, 35, 40, 50, 60, 70, and 80 mmHg), t...
