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Topic:Ex Vivo Study
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.
The propagation of induced tendon lesions in the equine superficial digital flexor tendon: an ex vivo study. 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...
IL-4 activates equine neutrophils and induces a mixed inflammatory cytokine expression profile with enhanced neutrophil chemotactic mediator release ex vivo. Neutrophils are potent contributors to the lung pathophysiological changes occurring in allergic airway inflammation, which typically involve T helper type 2 (Th2) cytokine overexpression. We have previously reported that equine pulmonary endothelial cells are activated by the Th2 cytokine IL-4 and express chemotactic factors for neutrophils after stimulation. We have further explored the possible mechanisms linking Th2-driven inflammation and neutrophilia by studying the effects of recombinant equine IL-4, a prototypical Th2 cytokine, on peripheral blood neutrophils (PBN) isolated from normal...
Distal interphalangeal articular cartilage assessment using low-field magnetic resonance imaging. The suitability of low-field magnetic resonance (MR) imaging for assessment of articular cartilage has been questioned, based on insufficient image quality. The purposes of this study were to describe the MR anatomy of the normal distal interphalangeal (DIP) cartilage, and to evaluate the sensitivity and accuracy of low-field MR imaging for identification of cartilage erosions that were created ex vivo. Imaging sequences included sagittal and dorsal multiple-oblique T1-weighted gradient-recalled echo (GRE) and sagittal dual echo sequences. In the thickest regions, normal cartilage appeared as ...
Fat-suppressed spoiled gradient-recalled imaging of equine metacarpophalangeal articular cartilage. 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...
Correlation of signal attenuation-based quantitative magnetic resonance imaging with quantitative computed tomographic measurements of subchondral bone mineral density in metacarpophalangeal joints of horses. To evaluate the ability of signal attenuation-based quantitative magnetic resonance imaging (QMRI) to estimate subchondral bone mineral density (BMD) as assessed via quantitative computed tomography (QCT) in osteoarthritic joints of horses. Methods: 20 metacarpophalangeal joints from 10 horse cadavers. Methods: Magnetic resonance (MR) images (dorsal and transverse T1-weighted gradient recalled echo [GRE] and dorsal T2*-weighted GRE fast imaging employing steady-state acquisition [T2*-FIESTA]) and transverse single-slice computed tomographic (CT) images of the joints were acquired. Magnetic res...
Debridement of cartilage lesions before autologous chondrocyte implantation by open or transarthroscopic techniques: a comparative study using post-mortem materials. We compared the quality of debridement of chondral lesions performed by four arthroscopic (SH, shaver; CU, curette; SHCU, shaver and curette; BP, bipolar electrodes) and one open technique (OPEN, scalpel and curette) which are used prior to autologous chondrocyte implantation (ACI). The ex vivo simulation of all five techniques was carried out on six juvenile equine stifle joints. The OPEN, SH and SHCU techniques were tested on knees harvested from six adult human cadavers. The most vertical walls with the least adjacent damage to cartilage were obtained with the OPEN technique. The CU and SHC...
Magnetic resonance signal changes during time in equine limbs refrigerated at 4 degrees C. 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...
Ex vivo serotype-specific transduction of equine joint tissue by self-complementary adeno-associated viral vectors. Cell transplantation for the treatment of joint disease is an important clinical tool. Genetic modification of cells before transplantation has shown enhanced healing. Ex vivo genetic modification of joint tissue cells with various adeno-associated virus (AAV) serotypes has not been investigated. The transduction efficiencies of self-complementary AAV serotypes (1-6 and 8) were determined in joint tissue containing chondrocytes and synoviocytes isolated from equine models. When comparing scAAV serotypes for efficient transduction ex vivo, in chondrocytes versus synoviocytes, serotypes 6 and 2,...
Use of computed tomography in standing position to identify guidelines for screw insertion in the distal phalanx of horses: an ex vivo study. To compare the precision of radiography and computed tomography (CT) preoperatively in the standing position for identification of guidelines for screw insertion in the distal phalanx, and to identify whether standing CT might improve operative time compared with preoperative radiographic planning. Methods: Experimental ex vivo study. Methods: Cadaveric equine thoracic limb pairs (n=10). Methods: Insertion of a 4.5 mm cortex screw in lag fashion into an intact distal phalanx was evaluated in 2 groups (n=10) of cadaveric equine thoracic limbs. In 1 group, the site, direction, and length of the ...
Ex vivo calibration and validation of in vivo equine bone strain measures. Data are required to confirm that strain gauges recording high bone strains in Thoroughbred racehorses provide an accurate record of bone strain. Objective: To test the accuracy and reliability of very high in vivo strain recordings made during fast exercise in Thoroughbred racehorses. Methods: Strains were recorded during exercise from rosette gauges implanted onto the mid-shaft dorsal cortex of each third metacarpal bone (MC3) in 6 yearling and 6 mature Thoroughbreds in a previous experiment. Bulk elastic modulus (E(US)) was calculated from ultrasound speed and single photon absorptiometry m...
Effects of long-term extracorporeal blood perfusion of the distal portion of isolated equine forelimbs on metabolic variables and morphology of laminar tissue. 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 ...
Isolation, establishment, and characterization of ex vivo equine melanoma cell cultures. Gray horses spontaneously develop metastatic melanomas that resemble human disease, and this is often accompanied with metastasis to other organs. Unlike in other species, the establishment of primary equine melanoma cultures that could be used to develop new therapeutic approaches has remained a major challenge. The purpose of the study was to develop a protocol for routine isolation and cultivation of primary equine melanocytes. Melanoma tissues were excised from 13 horses under local anesthesia, mainly from the perianal area. The melanoma cells were isolated from the melanoma tissue by seri...
MRI-based inverse finite element approach for the mechanical assessment of patellar articular cartilage from static compression test. The mechanical property of articular cartilage determines to a great extent the functionality of diarthrodial joints. Consequently, the early detection of mechanical and, thus, functional changes of cartilage is crucial for preventive measures to maintain the mobility and the quality of life of individuals. An alternative to conventional mechanical testing is the inverse finite element approach, enabling non-destructive testing of the tissue. We evaluated a method for the assessment of the equilibrium material properties of the patellar cartilage based on magnetic resonance imaging during pate...
The use of radial extracorporeal shockwave therapy in the treatment of urethral urolithiasis in the horse: a preliminary study. Radial extracorporeal shockwave therapy (ESWT) is widely used in equine practice for the treatment of orthopedic problems. However, its original use as a lithotripsy device in human and canine urology led us to postulate that it could be used as an alternative to the surgical treatment of urethral calculi in horses. Objective: Radial ESWT can easily and safely fragment calculi in the distal urethra of the horse. Methods: Two postmortem cases and 1 live case of obstructive urinary disease admitted at the Veterinary Teaching Hospital of Liege. Methods: A radial shockwave device was directly appl...
Isolation and culture of primary equine tracheal epithelial cells. Culture of airway epithelial cells is a useful model to investigate physiology of airway epithelia and airway disease mechanisms. In vitro models of airway epithelial cells are established for various species. However, earlier published method for isolation and culture of equine tracheal epithelial cells requires significant improvements. In this report, the development of a procedure for efficient isolation, characterization, culture, and passage of primary equine tracheal epithelial cells are described. Epithelial cells were isolated from adult equine trachea by exposing and stripping the mu...
Full-length and internally deleted forms of interleukin-7 are present in horse (Equus caballus) lymph node tissue. Horse IL-7 (HIL-7) cDNA was isolated from adult lymph node tissue by reverse transcription polymerase chain reaction (RT-PCR) using oligonucleotide primers based on horse genomic sequences (The Broad Institute). In addition, to the full-length (FL) 531bp reading frame encoding 176 amino acids, shorter open-reading frames of 477, 396 and 264bp were also amplified. Nucleotide sequence analysis of these RT-PCR products demonstrated they were homologous except the shorter species were missing internal sequences consistent with multiple RNA splicing events. Consequently, the shorter open-reading fr...
Evaluation of a laparoscopic technique for collection of serial full-thickness small intestinal biopsy specimens in standing sedated horses. To assess a technique for laparoscopic collection of serial full-thickness small intestinal biopsy specimens in horses. Methods: 13 healthy adult horses. Methods: In the ex vivo portion of the study, sections of duodenum and jejunum obtained from 6 horses immediately after euthanasia were divided into 3 segments. Each segment was randomly assigned to the control group, the double-layer hand-sewn closure group, or the endoscopic linear stapler (ELS) group. Bursting strength and bursting wall tension were measured and compared among groups; luminal diameter reduction at the biopsy site was compa...
Fracture configurations of the equine radius and tibia after a simulated kick. The objective of this postmortem study was to determine the fracture configurations of the equine radius and tibia after a simulated kick. Fracture configurations of 35 radii and 36 tibiae from 19 adult horses were evaluated after a simulated kick in an experimental ex-vivo study. The bones were dissected, the proximal and distal ends were embedded in resin, fixed horizontally and preloaded in compression, and a steel impactor, designed to simulate a shod equine hoof, was dropped from a height of three to six metres onto the diaphysis. The experiments were filmed with a high-speed camera (30,0...
Assessment of exercise-induced alterations in neutrophil function in horses. 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....
A surgical approach to the lateral compartment of the equine guttural pouch in the standing horse: modification of the forgotten “Garm technique”. The objective of this study was to evaluate the feasibility, efficacy and complications following lavage and drainage of the lateral compartment (LC) of the equine guttural pouch (GP) using a modified Garm's technique (MGT). In an ex vivo study (study 1), six cadaver heads were examined to assess the anatomical limits of the surgical approach and whether vital structures might be damaged. This was followed by an in vivo study (study 2) in which a lavage/drainage tube was placed for 3 days into each LC of four standing horses using the MGT. In both studies, the procedure offered direct access i...
Reactivity of equine airways–a study on precision-cut lung slices. A study was performed to evaluate the use of precision-cut lung slices (PCLS) for studies on the contraction of equine airways. Lungs of 10 horses were taken to prepare PCLS of approximately 250 microm from equine lung tissue using a special microtome. The lung slices were cultured and the enclosed small airways were monitored using a microscope with coupled digital camera, which was used to determine the airway luminal area and diameter from digital images. As indicated by the beating of the ciliated epithelium and reactivity of airways on methacholine challenge, the tissue slices were found ...
Ex vivo comparison of a novel tapered-sleeve and traditional full-limb transfixation pin cast for distal radial fracture stabilization in the horse. Distal radial fractures in adult horses are examples of long-bone fractures that are not always amenable to internal fixation. These fractures are often open, contaminated, severely comminuted, and located adjacent to the antebrachiocarpal joint. There have been few studies to improve upon the methods of stabilization of this type of fracture. External coaptation incorporating transfixation pins is one method that has been used to stabilize distal radial fractures in horses (1-3). The purpose of this preliminary study was to compare the load to failure in simulated weight-bearing of a novel ta...
In situ and ex vivo evaluation of an arthroscopic indentation instrument to estimate the health status of articular cartilage in the equine metacarpophalangeal joint. To evaluate an arthroscopic indentation instrument (Artscan 200) for assessment of the health status of equine articular cartilage. Methods: In vitro experiment using equine isolated proximal phalanx (P1) specimens. Methods: P1 specimens from 39 horses (aged 1.5-22 years). Methods: Reproducibility was tested by determination of the coefficient of variation (CV). Dynamic modulus and cartilage degeneration index (CDI) values were measured at 2 predefined sites (site 1, joint margin; site 2, joint center) to assess the accuracy and to evaluate the relation with surface integrity. Results: CV was ...
Third metacarpal bone mineral density assessment in the standing horse by dual X-ray absorptiometry: suitability, precision and accuracy. Bone mineral density (BMD) is correlated to mechanical properties of bone. In the horse, dual energy X-ray absorptiometry (DXA) has yet only been performed ex-vivo, but a new portable DXA device would be ideal for in-vivo BMD measurement. We explored field suitability, precision and accuracy of this device for in-vivo third metacarpal density assessment. Precision was analysed by calculating measurement variation under repeated measurement tests with (reproducibility) and without (repeatability) limb repositioning. Repeatability and reproducibility were tested ex-vivo, at the same time that in...
Ex vivo strength comparison of bioabsorbable tendon plates and bioabsorbable suture in a 3-loop pulley pattern for repair of transected flexor tendons from horse cadavers. To test the failure strength and energy of 2 bioabsorbable implants applied to transected deep digital flexor tendons (DDFT) from adult horses. Methods: Ex vivo biomechanical experiment. Methods: Twelve pairs of deep digital flexor tendons harvested from the forelimbs of fresh equine cadavers. Methods: Poly-L-lactic acid tendon plates were custom manufactured for application to the cylindrical surface of an adult equine deep digital flexor tendon. Twelve pairs of DDFTs were transected 2 cm distal to the insertion of the distal check ligament of the deep digital flexor tendon. One tendon of eac...
Magnetic resonance imaging findings in the equine deep digital flexor tendon and distal sesamoid bone in advanced navicular disease–an ex vivo study. We describe the abnormal magnetic resonance (MR) imaging findings in the deep digital flexor tendon (DDFT) and distal sesamoid bone in horses with radiographic changes compatible with navicular syndrome. Thirteen postmortem specimens were examined using a 1.5-T magnetic field, with spin echo (SE) T1-weighted, turbo SE (TSE) proton density-weighted (with and without fat saturation), and fat saturation TSE T2-weighted sequences. The limbs were then dissected to compare the MR findings with the gross assessment and histologic examination of the DDFT and distal sesamoid bones. Tendonous abnormalit...
Update on equine ICSI and cloning. 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 ...
In vivo pretreatment with PGG-glucan fails to alter cytokine mRNA expression of equine peripheral blood mononuclear cells exposed to endotoxin ex vivo. Twelve healthy horses were assigned to treatment or control groups. Treated horses received PGG-Glucan[ED-1] (1 mg/kg, IV) 24 hours prior to peripheral blood mononuclear cell (PBMC) isolation. PBMCs were isolated and incubated in the presence of lipopolysachharide (LPS). At 0, 6, 12, 24 and 48 hours messenger RNA (mRNA) was extracted. Reverse transcription polymerase chain reaction (PCR) was performed and cytokine mRNA expression for tumor necrosis factor alpha (TNFalpha), interleukin 1beta (IL-1beta), interleukin 10 (IL-10) and interferon gamma (IFN-gamma) determined using real time PCR. A si...
Optimization of assay conditions for leukotriene B4 synthesis by neutrophils or platelets isolated from peripheral blood of monogastric animals. 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...
Ex vivo investigation of the use of hydrothermal energy to induce chondrocyte necrosis in articular cartilage of the metacarpophalangeal and metatarsophalangeal joints of horses. To evaluate the use of hydrothermal ablation of articular cartilage for arthrodesis in horses through investigation of the effects of joint lavage with physiologic saline (0.9% NaCI) solution (80 degrees C) for various treatment times on chondrocyte viability in the articular cartilage of the metacarpophalangeal and metatarsophalangeal joints of cadaveric horse limbs. Sample Population-7 pairs of metacarpophalangeal and 8 pairs of metatarsophalangeal joints from 8 Thoroughbreds. Methods: The horses were euthanatized for reasons unrelated to musculoskeletal disease. On a random basis, 1 joint o...
