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.
Behroozi M, Graïc JM, Gerussi T.Diffusion-weighted Imaging (DWI) is an effective and state-of-the-art neuroimaging method that non-invasively reveals the microstructure and connectivity of tissues. Recently, novel applications of the DWI technique in studying large brains through imaging enabled researchers to gain insights into the complex neural architecture in different species such as those of (e.g., horses and rhinos), (e.g., bovids, swines, and cetaceans), and (e.g., felids, canids, and pinnipeds). Classical tract-tracing methods are usually considered unsuitable for ethical and practical reasons, in large animals...
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...
Grzeskowiak R, Schumacher J, Omidi O, Bowers K, Cassone LMC, Abedi R, Hespel AM, Mulon PY, Anderson DE.To assess a three-dimensional (3-D)-printed laryngeal clamp (LC) designed to enhance the anchoring of laryngeal prostheses at the cricoid cartilage. Methods: Ex vivo biomechanical study. Methods: A total of 22 equine larynges. Methods: Two experimental groups included larynges with standard prosthetic laryngoplasty (PL; n = 10) and larynges with prosthetic laryngoplasty modified with laryngeal clamps (PLLC; n = 10). All constructs underwent 3000 cycles of tension loading and a single tension to failure. Recorded biomechanical parameters included maximum load, actuator displacement, a...
de Souza AF, Pereira CAM, Costa C, Fürst A, Kümmerle JM, De Zoppa ALV. The aim of this study was to measure the mechanical properties and failure mode of an model of proximal interphalangeal arthrodesis in horses with different methods of proximal screw fixation using a locking compression plate. Methods: Pastern joint arthrodesis with a 3-hole 4.5-mm narrow locking compression plate was performed on 20 forelimbs of equine cadavers, randomized into four groups based on the fixation method and type of proximal screw used: unicortical cortex, bicortical cortex, unicortical locking, bicortical locking. Single-cycle axial compression testing was conducted unti...
Köhne M, Diel E, Packeiser EM, Böttcher D, Tönissen A, Unruh C, Goericke-Pesch S, Ulrich R, Sieme H.In the past, most research in equine reproduction has been performed in vivo but the use of in vitro and ex vivo models has recently increased. This study aimed to evaluate the functional stability of an ex vivo hemoperfused model for equine uteri with molecular characterization of marker genes and their proteins. In addition, the study validated the respective protein expression and the aptness of the software QuPath for identifying and scoring immunohistochemically stained equine endometrium. After collection, uteri (n = 12) were flushed with preservation solution, transported to the labor...
Bierau J, Cruz AM, Koch C, Manso-Diaz G, Büttner K, Staszyk C, Röcken M.Cone-beam computed tomography (CBCT) is regarded as a convenient and suitable alternative to conventional computed tomography. However, in the horse, the quality of obtained data sets needs to be evaluated. Therefore, the aim of this study was to compare the visibility and accessibility of clinically relevant anatomical structures displayed in CBCT and conventional multidetector computed tomography (MDCT). Unassigned: Twenty-nine limbs from horses euthanized for reasons unrelated to this study were used. Native and intraarticular contrast scans of the fetlock (CBCT vs. MDCT) were performed. Th...
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...
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 ...
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...
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 ...
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...
Fugazzola M, Nissinen MT, Jäntti J, Tuppurainen J, Plomp S, Te Moller N, Mäkelä JTA, van Weeren R.Strategies for articular cartilage repair need to take into account topographical differences in tissue composition and architecture to achieve durable functional outcome. These have not yet been investigated in the equine stifle. Objective: To analyse the biochemical composition and architecture of three differently loaded areas of the equine stifle. We hypothesise that site differences correlate with the biomechanical characteristics of the cartilage. Methods: Ex vivo study. Methods: Thirty osteochondral plugs per location were harvested from the lateral trochlear ridge (LTR), the distal int...
Himebaugh NE, Robertson JB, Weninger K, Gilger BC, Ekesten B, Oh A.The aim of this study was to assess the transmission of the ultraviolet (UV) radiation (200-400 nm) through intact enucleated globes of different species (dogs, cats, pigs, rabbits, horses, and humans) using spectrophotometry. Globes of cats (n = 6), dogs (n = 18), pigs (n = 10), rabbits (n = 6), horses (n = 10), and humans (n = 4) were analyzed. A 5-10 mm circular area of sclera and choroid from the posterior aspect of the globe was removed under a surgical microscope, leaving the retina intact in all species except the horse. Glass coverslips were added in horses and rabbits du...
Munsterman AS, VanderBroek AR, Kottwitz JJ, Watson VE.To compare a novel suture loop method for intestinal biopsies (SLB) with a two-layer, hand-sutured biopsy (HSB) technique in equine small and large intestines. Methods: Experimental, randomized, ex vivo study. Methods: Eight healthy adult horses. Methods: The duodenum, aboral jejunum, and ventral and dorsal large colon were harvested after euthanasia and divided into three sections each. The sections were randomized to SLB, HSB, or control (C, no biopsy) groups. Tissue samples were excised after placement of a suture loop formed by a 4S Roeder knot and oversewn with a Cushing pattern using bar...
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...
Mora-Pereira M, Boone L, Naskou M, Wooldridge A.To evaluate ex vivo angiogenesis of equine arterial rings in response to various growth media. Methods: Facial arteries were dissected from 11 horses post-euthanasia. Equine platelet lysate (ePL) was harvested from 6 horses. Methods: Arteries were exposed to endothelial growth media (EGM) + horse serum (HS) for first sprout (FS), vascular regression (VR), and (basement membrane matrix [Matrigel]) lysis (ML) evaluation. Additional rings supplemented with (1) EGM, (2) EGM + EDTA, (3) endothelial basal media (EBM), (4) EBM + HS, or (5) EBM + human VEGF were compared for vascular network area (VNA...
Jarry J, De Raeve Y, Dugdale A, Simon V, Vandeweerd JM.Severe ophthalmic conditions such as trauma, uveitis, corneal damage, or neoplasia can lead to eye removal surgery. Poor cosmetic appearance resulting from the sunken orbit ensues. The aim of this study was to demonstrate the feasibility of manufacturing a custom-made 3D-printed orbital implant made of biocompatible material for the enucleated horse and usable in conjunction to a corneoscleral shell. Blender, a 3D-image software, was used for prototype design. Twelve cadaver heads of adult Warmbloods were collected from the slaughterhouse. On each head, one eye was removed via a modified trans...
Pala S, Hänninen NE, Mohammadi A, Ebrahimi M, Te Moller NCR, Brommer H, René van Weeren P, Mäkelä JTA, Korhonen RK, Afara IO, Töyräs J....The aim of this study is to assess whether articular cartilage changes in an equine model of post-traumatic osteoarthritis (PTOA), induced by surgical creation of standard (blunt) grooves, and very subtle sharp grooves, could be detected with ex vivo T relaxation time mapping utilizing three-dimensional (3D) readout sequence with zero echo time. Grooves were made on the articular surfaces of the middle carpal and radiocarpal joints of nine mature Shetland ponies and osteochondral samples were harvested at 39 weeks after being euthanized under respective ethical permissions. T relaxation times ...
Pozor MA, Benson SM, Macpherson ML, Kelleman AA.Transplantation of stem cells into dysfunctional testes is currently being investigated as a therapeutic option for men and stallions with advanced testicular degeneration. This series of "proof of concept" studies aimed to identify a safe and efficient method of inducing severe testicular degeneration to create an optimal equine recipient model for intratesticular stem cell transplantation (SCT). Two ex vivo and two in vivo experiments were conducted. At first, forty testes obtained from castrations were used to identify an effective therapeutic ultrasound (TUS) device and the protocol for ...
Gandini M, Cerullo A, Gallo L, Iussich S, Minoli L, Giusto G.To describe a technique for a side-to-side jejunocecal anastomosis in horses using radiofrequency thermofusion (TF) of the intestines supported by a Cushing oversew and to compare this anastomosis to handsewn and stapled techniques. Methods: Ex vivo study. Methods: Intestinal tracts from 24 slaughtered horses. Methods: A radiofrequency device was used to perform a jejunocecal anastomosis (Group RFA). The construction time and bursting pressure of this construct were compared with those of a hand-sewn double layer (Group HS) and stapled anastomoses (Group ST) without oversew of the staple line....
Swagemakers JH, Van Daele P, Mageed M.Cervical spinal nerve insult can be a sequel of osteoarthropathy in horses due to enlargement of the articular processes (AP). Objective: To describe the percutaneous endoscopic cervical foraminotomy procedure in horses. Methods: Ex vivo experimental study and clinical case report. Methods: The technique was performed in three equine cadavers and in two clinical cases with history of forelimb lameness located in the caudal cervical region. Briefly, the horse was positioned in lateral recumbency, with the affected AP joint uppermost. Under fluoroscopic guidance, the endoscopic instruments (Ø 6...
De Gasperi D, Guo D, Guo D, Lu Y, Brounts SH.To develop and describe a minimally invasive, ultrasound-guided, percutaneous technique for the desmotomy of equine palmar/plantar annular ligaments (PALs) using a transecting thread. Methods: Ex vivo study. Methods: Twenty-one normal equine distal limb specimens. Methods: Under ultrasonographic guidance, a surgical thread was percutaneously placed around the PAL through 2 skin punctures (proximal and distal) using a 18 gauge spinal needle in equine limbs (11 forelimbs, 10 hindlimbs) with normal PALs. The ligament was transected by a back-and-forth motion of the thread until the loop emerged...
Aparicio IM, Rojo-Domínguez P, Castillejo-Rufo A, Peña FJ, Tapia JA.Despite its importance in somatic cells and during spermatogenesis, little is known about the role that autophagy may play in ejaculated spermatozoa. Our aim was to investigate whether the molecular components of autophagy, such as microtubule-associated protein 1 light chain 3 (LC3), are activated in stallion spermatozoa during the capacitation and acrosome reaction and if this activation could modulate these biological processes. To analyze the autophagy turnover, LC3I and LC3II proteins were assessed by western blotting, and the ratio between both proteins (LC3II/LC3I) was calculated. In so...
Granello ME, Weatherall KM, Lutter JD, Beard WL. The aim of this study was to compare the deflection distance, deflection angle and pre-loading sub-bandage pressure of a transverse, mid-metacarpal fracture in an equine cadaver limb when loaded in bending while stabilized with either a Robert Jones Bandage (RJB) or a single-stack bandage (SS), each containing two orthogonal splints. Methods: In an ex vivo experimental study, forelimb pairs from eight horses were collected, and one forelimb from each pair was assigned to either the SS or RJB groups. A transverse osteotomy was performed at the midpoint of the metacarpus. After bandaging, ...
Depuydt E, Chiers K, Van Hecke L, Saunders J, Martens A, Pille F, Spaas JH.Injuries to equine tendons and ligaments are career-compromising, causing reduced performance and premature retirement. Promising treatment alternatives have been investigated in the field of mesenchymal stem cells (MSCs). In this study, the tissue adherence and protein expression of tenogenic primed mesenchymal stem cells (tpMSCs) after administration to ex vivo tendon and ligament explants is investigated. First, collagen type I (COL I) and smooth muscle actin (SMA) expression was assessed in cytospins prepared from native MSCs and tpMSCs. Second, equine superficial digital flexor tendon and...
Ząbek T, Witarski W, Szmatoła T, Sawicki S, Mrozowicz J, Samiec M.Epigenetic mechanisms of gene regulation are important for the proper differentiation of cells used for therapeutic and regenerative purposes. The primary goal of the present study was to investigate the impacts of 5-aza-2' deoxycytidine (5-AZA-dc)- and/or trichostatin A (TSA)-mediated approaches applied to epigenomically modulate the ex vivo expanded equine chondrocytes maintained in monolayer culture on the status of chondrogenic cytodifferentiation at the transcriptome level. The results of next-generation sequencing of 3' mRNA-seq libraries on stimulated and unstimulated chondrocytes of th...
Seitz-Cherner E, Bauck AG, Denagamage T, Freeman DE.To compare single-layer anastomoses (modified continuous Lembert [mod-CL] and modified-interrupted Lembert [mod-IL]) and a 2-layer simple continuous anastomosis (2 L; seromuscular and mucosal) for jejunojejunal anastomoses in equine cadavers and to compare ex vivo to in vivo time to complete the anastomosis and stoma size with a mod-IL pattern. Methods: Measurements in jejunum from cadaver and anesthetized horses. Methods: Ten live horses and 18 equine cadavers. Methods: Time to complete anastomosis, bursting pressures (BP), leakage sites, and anastomotic index (size ratio of anastomotic lume...
Hall NP, Heaton KT, Farnsworth KD, Ragle CA.This report evaluates the use of 4.5- and 5.5-mm cannulated drill bits for articular cartilage removal from the proximal interphalangeal joints of equine cadaver limbs. Limbs from 8 equine cadavers, all with normal proximal interphalangeal joints. Proximal interphalangeal joints of 32 limbs from 8 equine cadavers were drilled using either 4.5- or 5.5-mm cannulated drill bits. Pastern joints were then disarticulated, and intra-articular drilling was evaluated by visual inspection. Post-drilling evaluation revealed complete intra-articular drilling occurred in all 32 joints. Canulated 4.5- and 5...
Miagkoff L, Girard CA, St-Jean G, Richard H, Beauchamp G, Laverty S.Osteoarthritis (OA) is a major cause of equine lameness. Cannabinoid (CB) receptors are now considered to be promising therapeutic targets in human rheumatology for pain and inflammation, however, little is known about the equine endocannabinoid system. Objective: The primary goal was to assess the presence and expression pattern of CB1 and CB2 in the synovium of healthy joints. A secondary goal was to explore the relationship between the CB expression, degree of synovitis and OA pathology. Methods: Ex vivo experimental study. Methods: Metacarpophalangeal joints (n = 25) from a tissue bank w...
Ząbek T, Witarski W, Semik-Gurgul E, Szmatoła T, Kowalska K, Samiec M.Ex vivo expansion of chondrocytes in monolayer (ML) culture for therapeutic purposes is burdened with difficulties related to the loss of cartilaginous phenotype. Epigenetic mechanisms responsible for regulation of gene expression are believed to underlie chondrocyte dedifferentiation. We have inspected the relevance of DNA methylation alterations for passage-related differential expression of NFATC1 gene involved in hard connective tissue turnover and development, NADSYN1 influencing redox metabolism, and JAK3 - an important driver of inflammation. We have assessed relative amount of transcri...
Sleutjens J, Voorhout G, Van Der Kolk JH, Wijnberg ID, Back W.In dressage, the head and neck position has become an issue of concern as certain extreme positions may imply a welfare risk for the horse. In man, extension and flexion of the cervical spine cause a decrease and increase in intervertebral foramina dimensions, respectively. However, in horses, the influence of flexion and extension on foramina dimensions and its possible interference with peripheral nerve functioning remains unknown. Objective: To determine the effect of ex vivo flexion and extension on intervertebral foramina dimensions in the equine cervical spine. Methods: Computed tomograp...
Veraa S, Bergmann W, Wijnberg ID, Back W, Vernooij H, Nielen M, van den Belt AM.Morphology of the equine cervical intervertebral disc is different from that in humans and small companion animals and published imaging data are scarcely available. The objectives of this exploratory, methods comparison study were (a) to describe MRI features of macroscopically nondegenerated and degenerated intervertebral discs (b) to test associations between spinal location and macroscopic degeneration or MRI-detected annular protrusion and between MRI-detected annular protrusion and macroscopic degeneration, and (c) to define MRI sequences for characterizing equine cervical intervertebral...
Van Cleemput J, Poelaert KCK, Laval K, Van den Broeck W, Nauwynck HJ.The horse's respiratory tract daily encounters a plethora of respirable hazards including air pollutants, mycotoxins and airborne pathogens. To date, the precise effect of air pollution and mycotoxins on respiratory epithelial integrity and subsequent pathogen invasion in the horse has not been studied. Here, diesel exhaust particles (DEP) and three major mycotoxins (deoxynivalenol [DON], aflatoxin B1 [AFB1] and fumonisin B1 [FB1]) were applied to the apical surfaces of both ex vivo respiratory mucosal explants and in vitro primary equine respiratory epithelial cells (EREC) cultivated at the a...
Marlo TL, Giuliano EA, Sharma A, Mohan RR.To develop an ex vivo equine corneal organ culture model. Specifically, to assess the equine cornea's extracellular matrix and cellularity after 7 days using two different culture techniques: either (i) immersion system or (ii) air/liquid interface system, to determine the best ex vivo equine corneal model. Methods: Fourteen healthy equine corneas of various breeds. Methods: Equine corneas with 2 mm of perilimbal sclera were freshly harvested from 7 horses undergoing humane euthanasia. One corneal-scleral ring (CSR) from each horse was randomly placed in the (i) immersion condition organ cultu...
Miagkoff L, Girard CA, St-Jean G, Richard H, Beauchamp G, Laverty S.Osteoarthritis (OA) is a major cause of equine lameness. Cannabinoid (CB) receptors are now considered to be promising therapeutic targets in human rheumatology for pain and inflammation, however, little is known about the equine endocannabinoid system. Objective: The primary goal was to assess the presence and expression pattern of CB1 and CB2 in the synovium of healthy joints. A secondary goal was to explore the relationship between the CB expression, degree of synovitis and OA pathology. Methods: Ex vivo experimental study. Methods: Metacarpophalangeal joints (n = 25) from a tissue bank w...
De Lasalle J, Alexander K, Olive J, Laverty S.A better understanding of imaging characteristics of equine stifle osteoarthritis (OA) may allow earlier detection and improve prognosis. Objectives of this ex vivo, prospective, methods comparison study were to (1) describe the location and severity of naturally acquired OA lesions in the equine stifle using ultrasound (US), radiography (XR), computed tomography (CT), and macroscopic evaluation (ME); (2) compare the diagnostic performance of each imaging modality with ME; and (3) describe subchondral bone mineral density (BMD) in equine stifle joints with OA using CT. Radiographic, CT, and US...
Anderson SL, Singh B.Horses that develop colitis invariably exhibit signs of a systemic inflammatory response syndrome (SIRS). A significant contributor to the development of SIRS in human subjects is delayed neutrophil apoptosis, but this has not been specifically studied in horses. Objective: To determine the occurrence of ex vivo neutrophil apoptosis and its contribution to the development of SIRS in an equine colitis model. Methods: Experiment using a colitis model. Methods: Neutrophils were isolated before and after the induction of colitis using an oligofructose overdose model, placed into culture for 12 h o...
Laverty S, Okouneff S, Ionescu M, Reiner A, Pidoux I, Webber C, Rossier Y, Billinghurst RC, Poole AR.Articular osteochondrosis (OCD) occurs in both man and animals. The etiology remains to be determined. Studies of OCD lesions in animals may provide clues as to its pathogenesis. The aim of our study was to determine whether there was evidence for increased degradation namely proteoglycan (PG) release and type II collagen cleavage in articular cartilage harvested from OCD lesions. We examined ex vivo explants at post-mortem from equine OCD lesions and macroscopically normal site and age matched cartilage. These were cultured over a 10 day period in serum-free medium. Type II collagen cleavage ...
Dubois MS, Morello S, Rayment K, Markel MD, Vanderby R, Kalscheur VL, Hao Z, McCabe RP, Marquis P, Muir P.Articular stress fracture arising from the distal end of the third metacarpal bone (MC3) is a common serious injury in Thoroughbred racehorses. Currently, there is no method for predicting fracture risk clinically. We describe an ex-vivo biomechanical model in which we measured subchondral crack micromotion under compressive loading that modeled high speed running. Using this model, we determined the relationship between subchondral crack dimensions measured using computed tomography (CT) and crack micromotion. Thoracic limbs from 40 Thoroughbred racehorses that had sustained a catastrophic in...
Goulet C, Olive J, Rossier Y, Beauchamp G.Early radiographic diagnosis of acute laminitis is important for treatment planning and prognosis in horses. While four histopathologic layers make up the hoof wall, only two layers are distinguished on digital radiographs of horses' hooves. Objectives of this descriptive study were to determine which macroscopic layers correspond to the two radiographically visible layers, and to describe radiographic layer measurements in a group of clinically sound Warmblood horses. Twelve ex vivo equine hooves were examined at three different levels of the dorsal hoof wall. The four macroscopic layers were...
Bolen G, Haye D, Dondelinger R, Busoni V.When ex vivo magnetic resonance (MR) imaging studies are undertaken, specimen conservation should be taken into account when interpreting MR imaging results. The purpose of this study was to assess MR changes during time in the anatomic structures of the equine digit on eight cadaver limbs stored at 4 degrees C. The digits were imaged within 12 h after death and then after 1, 2, 7, and 14 days of refrigeration. After the last examination, four feet were warmed at room temperature for 24 h and reimaged. Sequences used were turbo spin echo (TSE) T1, TSE T2, short tau inversion recovery (STIR), a...
Bosch G, Lameris MC, van den Belt AJ, Barneveld A, van Weeren PR.Clinical tendon lesions usually enlarge during the first days to weeks after sustaining the injury due to enzymatic and biomechanical influences. Limiting this enlargement would positively influence prognosis related to lesion size. Objective: To investigate the effect of cyclic loading on the propagation of enzymatically and physically induced tendon lesions and to assess the effect of immobilisation thereon in an ex vivo model. Methods: Equine cadaver limbs with either physically or collagenase-induced SDFT lesions were placed in a pneumatic loading device. Groups consisted of unloaded limbs...
Duesterdieck-Zellmer KF, Larson MK, Plant TK, Sundholm-Tepper A, Payton ME.OBJECTIVE To measure penetration efficiencies of low-level laser light energy through equine skin and to determine the fraction of laser energy absorbed by equine digital flexor tendons (superficial [SDFT] and deep [DDFT]). SAMPLE Samples of skin, SDFTs, and DDFTs from 1 metacarpal area of each of 19 equine cadavers. PROCEDURES A therapeutic laser with wavelength capabilities of 800 and 970 nm was used. The percentage of energy penetration for each wavelength was determined through skin before and after clipping and then shaving of hair, through shaved skin over SDFTs, and through shaved skin,...
Reisinger N, Schaumberger S, Nagl V, Hessenberger S, Schatzmayr G.Laminitis is one of the most common diseases in horses. It is not only painful for the animal, but also has a significant financial impact on the equine industry. This multifactorial disease affects the connective tissue of the hoof. However, the pathogenesis of laminitis is still not fully understood. Endotoxins, also known as lipopolysaccharides (LPS), and bacterial exotoxins seem to play an important role during the development of laminitis. The aim of our study was to investigate the effect of increasing LPS concentrations (0, 2.5, 5, 10, and 100 μg/mL) on cell viability of isolated epide...
Ledbetter EC, Irby NL, Kim SG.To describe in vivo corneal confocal microscopy of horses with fungal keratitis and correlate findings with clinical, histopathological, and microbiological evaluations of clinical cases and an ex vivo experimental equine fungal keratitis model. Methods: A total of 12 horses with naturally-acquired fungal keratitis and ex vivo equine corneas experimentally infected with clinical fungal isolates. Methods: Horses with naturally-acquired fungal keratitis were examined with a modified Heidelberg Retina Tomograph II and Rostock Cornea Module. Confocal microscopy images of clinical isolates of Asper...
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...
Jones KE.The thoracolumbar region is clinically important in horses; however, the link between joint mobility and bony joint morphology has not been tested quantitatively. Objective: To establish which aspects of vertebral morphology correlate with ex vivo range of motion in the thoracolumbar region of Equus caballus, and demonstrate methodologies for linking vertebral form and function. Methods: Morphometric study of osteological specimens. Methods: A digital model was created of a disarticulated thoracolumbar region to examine bone-to-bone interactions during in silico bending. Linear measurements a...
Patan B, Budras KD, Licka TF.To establish an ex vivo model of blood perfusion in the distal portion of isolated equine forelimbs that closely represents the in vivo situation in the laminar tissue of the hoof. Methods: 18 forelimbs collected from 9 healthy adult horses following slaughter at a licensed abattoir. Methods: The distal portion of isolated equine forelimbs from 9 horses were perfused under physiologic conditions over a period of 6, 8, and 10 hours with autologous blood. To determine cell viability in perfused tissues, indicators for metabolism (lactate generation and glucose and oxygen consumption) as well as ...
Ziegler A, Everett H, Hamza E, Garbani M, Gerber V, Marti E, Steinbach F.Dendritic cells are professional antigen-presenting cells that play an essential role in the initiation and modulation of T cell responses. They have been studied widely for their potential clinical applications, but for clinical use to be successful, alternatives to xenogeneic substances like fetal bovine serum (FBS) in cell culture need to be found. Protocols for the generation of dendritic cells ex vivo from monocytes are well established for several species, including horses. Currently, the gold standard protocol for generating dendritic cells from monocytes across various species relies u...
Barton MH, Paske E, Norton N, King D, Giguère S, Budsberg S.Two firocoxib preparations for oral use are approved for use in animals in many countries: a chewable canine tablet and an equine paste. In order to reduce costs, many veterinarians use the canine product in horses even though this is an off-label use of the preparation. Objective: To determine the relative efficacy of 2 commercially available firocoxib products to inhibit prostaglandin E₂ (PGE2) synthesis after oral dosing in horses. Methods: A crossover design using 8 adult horses (n = 4 for each preparation during each treatment period). Body weight range 532-614 kg. Methods: Horses recei...
Knych HK, Arthur RM, McKemie DS, Seminoff K, Hamamoto-Hardman B, Kass PH.Phenylbutazone (PBZ) is a potent mon-steroidal anti-inflammatory drug used commonly in performance horses. The objectives of the current study were to describe blood and urine concentrations and the pharmacokinetics of PBZ and its metabolites following intravenous (IV) and oral administration and to describe the duration of pharmacodynamic effect. To that end, 17 horses received an IV administration and 18 horses an oral administration of 2 g of PBZ. Blood and urine samples were collected prior to and for up to 96 hours post drug administration. Whole blood samples were collected at various t...
Bolen GE, Haye D, Dondelinger RF, Massart L, Busoni V.To assess the impact of cycles of freezing and thawing on magnetic resonance (MR) images (obtained by use of a 3-T magnet) of equine feet examined ex vivo. Methods: 9 forelimbs from 9 horse cadavers. Methods: 9 forefeet underwent MR imaging first at ambient temperature within 12 hours after the horses' death and then after each freezing-thawing cycle. Three digits underwent freezing and thawing (at 4°C for 36 hours) 2 times, 3 digits underwent freezing and thawing (at 4°C for 36 hours) once and rescanning after 24 hours at ambient temperature, and 3 digits underwent freezing and thawing at a...
Nelson BB, Mäkelä JTA, Lawson TB, Patwa AN, Barrett MF, McIlwraith CW, Hurtig MB, Snyder BD, Moorman VJ, Grinstaff MW, Goodrich LR, Kawcak CE.Cationic agent contrast-enhanced computed tomography (cationic CECT) characterizes articular cartilage ex vivo, however, its capacity to detect post-traumatic injury is unknown. The study objectives were to correlate cationic CECT attenuation with biochemical, mechanical and histological properties of cartilage and morphologic computed tomography (CT) measures of bone, and to determine the ability of cationic CECT to distinguish subtly damaged from normal cartilage in an in vivo equine model. Mechanical impact injury was initiated in equine femoropatellar joints in vivo to establish subtle ...
Bowser JE, Elder SH, Pasquali M, Grady JG, Rashmir-Raven AM, Wills R, Swiderski CE.Hereditary equine regional dermal asthenia (HERDA) is an autosomal recessive disorder of Quarter Horses characterised by skin fragility. Horses with HERDA have a missense mutation in peptidyl-prolyl cis-trans isomerase B (PPIB), which encodes cyclophilin B and alters folding and post translational modifications of fibrillar collagen. Objective: The study aimed to test the hypothesis that tendons, ligaments and great vessels, which, like skin, are rich in fibrillar collagen, will also have abnormal biomechanical properties in horses with HERDA. Methods: Ex vivo biomechanical study comparing...
Olive J, D'Anjou MA, Girard C, Laverty S, Theoret C.The purpose was to evaluate the capacity of 1.5 T magnetic resonance (MR) imaging to assess articular cartilage in racehorses with naturally occurring metacarpophalangeal joint osteoarthritis. A sagittal, three-dimensional spoiled gradient-recalled echo (SPGR) with fat saturation (FS) sequence was acquired ex vivo on 20 joints. Following joint dissection, specific areas on the third metacarpal condyle were designated for subsequent sampling for histologic cartilage thickness measurement and modified Mankin scoring. Cartilage thickness was measured and cartilage signal intensity was also graded...
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...
Donovan DC, Jackson CA, Colahan PT, Norton NN, Clapper JL, Moore JN, Hurley DJ.To evaluate the effects of a standardized exercise test to exhaustion in horses on leukocyte function ex vivo. Methods: 6 Thoroughbred geldings. Methods: Blood samples were obtained from each horse before exercise; at exhaustion (termed failure); and at 2, 6, 24, 48, and 72 hours after exercise to evaluate hematologic changes, rate of leukocyte apoptosis, and leukocyte production of reactive oxygen species (ROS) ex vivo. To assess leukocyte function, leukocyte ROS production in response to stimulation with lipopolysaccharide, peptidoglycan, zymosan, and phorbol myristate acetate was evaluated....
Da Costa Gómez TM, Radtke CL, Kalscheur VL, Swain CA, Scollay MC, Edwards RB, Santschi EM, Markel MD, Muir P.To determine whether bone microcracks are altered after application of focused and radial extracorporeal shock wave therapy (ESWT) to the equine distal limb. Methods: An ex vivo experimental model. Methods: A contralateral limb specimen was obtained from 11 Thoroughbred racehorses with a unilateral catastrophic injury. Distal limb specimens were also obtained from 5 non-racing horses. Methods: Three separate skin-covered bone segments were obtained from the mid-diaphysis of the metacarpus (MC3) or metatarsus (MT3). Focused (9,000 shockwaves, 0.15 mJ/mm2, 4 Hz) and radial (9,000 shockwaves, 0.1...
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...
Fugazzola M, Nissinen MT, Jäntti J, Tuppurainen J, Plomp S, Te Moller N, Mäkelä JTA, van Weeren R.Strategies for articular cartilage repair need to take into account topographical differences in tissue composition and architecture to achieve durable functional outcome. These have not yet been investigated in the equine stifle. Objective: To analyse the biochemical composition and architecture of three differently loaded areas of the equine stifle. We hypothesise that site differences correlate with the biomechanical characteristics of the cartilage. Methods: Ex vivo study. Methods: Thirty osteochondral plugs per location were harvested from the lateral trochlear ridge (LTR), the distal int...
