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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.
Ex vivo biomechanical stability of 5 cricoid-suture constructs for equine laryngoplasty. To determine the biomechanical properties of 5 suture constructs in the equine cricoid under cyclic loading and load to failure testing. Methods: Ex vivo study. Methods: Seventy-five equine cadaver larynges. Methods: Each larynx was implanted with 1 of 5 cricoid-suture constructs. The standard laryngoplasty, where a suture is passed once through the cricoid, including its caudal edge, was used in 2 constructs: 1 with 5 USP Ethibond (ES) and 1 with 2 mm Fibertape (FS). In the third construct, the 2 mm Fibertape was passed twice through the cricoid including its caudal edge (Double Loop-DL). Con...
In vitro comparison of 3 techniques of prosthesis attachment to the muscular process of the equine arytenoid cartilage. To determine the degree and ease of arytenoid abduction achieved with abaxial placement of a FASTak II suture anchor compared to 2 suture patterns with different directions of insertion-caudomedial to craniolateral and medial to lateral. Methods: Ex vivo experimental. Methods: Cadaveric larynges from 10 Standardbred racehorses. Methods: Each larynx was sequentially instrumented with all 3 arytenoid suture attachment in random order: (1) abaxial placement of a FASTak II suture, (2) caudomedial to craniolateral suture, and (3) medial to lateral suture placement. Each construct was abducted at 5N...
Minicircle Mediated Gene Delivery to Canine and Equine Mesenchymal Stem Cells. Gene-directed tissue repair offers the clinician, human or veterinary, the chance to enhance cartilage regeneration and repair at a molecular level. Non-viral plasmid vectors have key biosafety advantages over viral vector systems for regenerative therapies due to their episomal integration however, conventional non-viral vectors can suffer from low transfection efficiency. Our objective was to identify and validate in vitro a novel non-viral gene expression vector that could be utilized for ex vivo and in vivo delivery to stromal-derived mesenchymal stem cells (MSCs). Minicircle plasmid DNA v...
Exogenous lactobacilli mitigate microbial changes associated with grain fermentation (corn, oats, and wheat) by equine fecal microflora ex vivo. Cereal grains are often included in equine diets. When starch intake exceeds foregut digestion starch will reach the hindgut, impacting microbial ecology. Probiotics (e.g., lactobacilli) are reported to mitigate GI dysbioses in other species. This study was conducted to determine the effect of exogenous lactobacilli on pH and the growth of amylolytic and lactate-utilizing bacteria. Feces were collected from 3 mature geldings fed grass hay with access to pasture. Fecal microbes were harvested by differential centrifugation, washed, and re-suspended in anaerobic media containing ground corn, whe...
Corrigendum: Unfractionated and Low-Molecular-Weight Heparin and the Phosphodiesterase Inhibitors, IBMX and Cilostazol, Block Ex Vivo Equid Herpesvirus Type-1-Induced Platelet Activation. [This corrects the article on p. 99 in vol. 3, PMID: 27909693.].
Comparison of 3 suture closure techniques for pelvic flexure enterotomy in equine cadaveric large colon. To compare a 2-layer closure with suture line reversal for a pelvic flexure enterotomy to 1-layer and traditional 2-layer hand sewn closures. Methods: Ex vivo, simple randomized study. Methods: Large colon segments from adult horses (n = 18). Methods: Pelvic flexures were harvested from 18 horses and randomly assigned to 1 of 3 closure techniques (n = 6 per technique). A 10-cm enterotomy was made in each pelvic flexure and closed with the assigned technique. Closure time, luminal diameter via contrast radiographs, and bursting pressure were recorded for each specimen and compared betwe...
Multimodality scoring of chondral injuries in the equine fetlock joint ex vivo. We investigate the potential of a prototype multimodality arthroscope, combining ultrasound, optical coherence tomography (OCT) and arthroscopic indentation device, for assessing cartilage lesions, and compare the reliability of this approach with conventional arthroscopic scoring ex vivo. Areas of interest (AIs, N = 43) were selected from equine fetlock joints (N = 5). Blind-coded AIs were independently scored by two equine surgeons employing International Cartilage Repair Society (ICRS) scoring system via conventional arthroscope and multimodality arthroscope, in which high-frequency ul...
Effects of various antiplatelet drugs on ex vivo platelet activation induced by equine herpesvirus type 1. OBJECTIVE To evaluate the effects of treatment of horses with standard platelet inhibitors on ex vivo inhibition of platelet activation by equine herpesvirus type I (EHV-I). ANIMALS II healthy adult horses. PROCEDURES In a double-blinded, placebo-controlled crossover study, horses were treated orally for 5 days with theophylline (5 mg/kg, q 12 h), pentoxifylline (10 mg/kg, q 12 h), clopidogrel bisulfate (4 mg/kg, q 24 h), acetylsalicylic acid (20 mg/kg, q 24 h), or placebo. Horses received all treatments, each separated by a 3-week washout period. Platelet-rich plasma was prepared from citrate...
The effect of free and carrier-bound cortisol on equine neutrophil function. Cortisol is a key anti-inflammatory hormone that increases in bacterial sepsis and circulates predominantly bound to cortisol binding globulin (CBG). Only unbound cortisol was believed to be biologically active, but recent evidence suggests that CBG-bound cortisol also regulates inflammation. The objective of this study was to evaluate the effects of free and CBG-bound cortisol on equine neutrophil function ex vivo. We hypothesized that CBG would enhance cortisol-mediated suppression of neutrophil pro-inflammatory responses. Neutrophils isolated from 8 foals and 6 adult horses were exposed to ...
Unfractionated and Low-Molecular-Weight Heparin and the Phosphodiesterase Inhibitors, IBMX and Cilostazol, Block Ex Vivo Equid Herpesvirus Type-1-Induced Platelet Activation. Equid herpes virus type-1 (EHV-1) is a major pathogen of horses, causing abortion storms and outbreaks of herpes virus myeloencephalopathy. These clinical syndromes are partly attributed to ischemic injury from thrombosis in placental and spinal vessels. The mechanism of thrombosis in affected horses is unknown. We have previously shown that EHV-1 activates platelets through virus-associated tissue factor-initiated thrombin generation. Activated platelets participate in thrombus formation by providing a surface to localize coagulation factor complexes that amplify and propagate thrombin genera...
Equine dendritic cells generated with horse serum have enhanced functionality in comparison to dendritic cells generated with fetal bovine serum. 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...
Ex vivo penetration of low-level laser light through equine skin and flexor tendons. 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,...
Development of a novel ex vivo equine corneal model. 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...
Ex Vivo Computed Tomographic Evaluation of Morphology Variations in Equine Cervical Vertebrae. Diagnostic imaging is one of the pillars in the clinical workup of horses with clinical signs of cervical spinal disease. An improved awareness of morphologic variations in equine cervical vertebrae would be helpful for interpreting findings. The aim of this anatomic study was to describe CT variations in left-right symmetry and morphology of the cervical and cervicothoracic vertebrae in a sample of horses. Postmortem CT examinations of the cervical spine for horses without congenital growth disorders were prospectively and retrospectively recruited. A total of 78 horses (27 foals, 51 mature h...
Release kinetics of tumor necrosis factor-α and interleukin-1 receptor antagonist in the equine whole blood. Horses are much predisposed and susceptible to excessive and acute inflammatory responses that cause the recruitment and stimulation of polymorphnuclear granulocytes (PMN) together with peripheral blood mononuclear cells (PBMC) and the release of cytokines. The aim of the study is to develop easy, quick, cheap and reproducible methods for measuring tumor necrosis factor alpha (TNF-α) and interleukin-1 receptor antagonist (IL-1Ra) in the equine whole blood cultures ex-vivo time- and concentration-dependently. Results: Horse whole blood diluted to 10, 20 and 50 % was stimulated with lipopolysa...
Comparisons Among Radiography, Ultrasonography, and Computed Tomography for Ex Vivo Characterization of Stifle Osteoarthritis in the Horse. 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...
Neutrophil apoptosis is delayed in an equine model of colitis: Implications for the development of systemic inflammatory response syndrome. 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...
Ex Vivo Mechanical Evaluation of a Sternal ZipFix(®) Implant for Prosthetic Laryngoplasty in Horses. To evaluate the properties of a ZipFix(®) (ZipFix) implant in equine laryngeal cartilages. Methods: Ex vivo biomechanical study. Methods: Equine arytenoid (n=36) and cricoid cartilages (n=18). Methods: Suture bites were placed in arytenoid or cricoid cartilages using a ZipFix(®) implant or a single strand of USP 5 braided polyester (TiCron™), and arytenoid and cricoid cartilages were separately subjected to single load to failure (25 N preload) or cyclic loading for 1,000 cycles, followed by single load to failure. Load, distraction, and stiffness were recorded. Results: Four arytenoid-Zip...
Genetic Evolution during the development of an attenuated EIAV vaccine. The equine infectious anemia virus (EIAV) vaccine is the only attenuated lentiviral vaccine applied on a large scale that has been shown to be effective in controlling the prevalence of EIA in China. This vaccine was developed by successive passaging of a field-isolated virulent strain in different hosts and cultivated cells. To explore the molecular basis for the phenotype alteration of this vaccine strain, we systematically analyzed its genomic evolution during vaccine development. Results: Sequence analysis revealed that the genetic distance between the wild-type strain and six representati...
Ex vivo comparison of the giant and transfixing knot in equine open and closed castration. In equine castration, application of a ligature on the spermatic cord to prevent complications such as haemorrhage and evisceration has been reported with controversial results. Characteristics of commonly used knots have not been studied. Objective: To compare the modified transfixing and giant knots and the emasculator in open and closed equine castration techniques. Methods: Ex vivo experiment. Methods: A total of 144 testicles were randomly assigned to 2 groups for open or closed castration. Both groups were divided into 3 subgroups of 18 specimens each: emasculator only, emasculator plus ...
Effect of various testing conditions on results for a handheld reference point indentation instrument in horses. To compare results obtained with a handheld reference point indentation instrument for bone material strength index (BMSi) measurements in the equine third metacarpal bone for various testing conditions. Methods: 24 third metacarpal bones. Methods: Third metacarpal bones from both forelimbs of 12 horses were obtained. The dorsal surface of each bone was divided into 6 testing regions. In vivo and ex vivo measurements of BMSi were obtained through the skin and on exposed bone, respectively, to determine effects of each testing condition. Difference plots were used to assess agreement between BM...
Concentration Dependent Influence of Lipopolysaccharides on Separation of Hoof Explants and Supernatant Lactic Acid Concentration in an Ex Vivo/In Vitro Laminitis Model. 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...
Endobronchial Ultrasound Reliably Quantifies Airway Smooth Muscle Remodeling in an Equine Asthma Model. Endobronchial ultrasonography (EBUS) revealed differences in the thickness of the layer representing subepithelial tissues (L2) between human asthmatics and controls, but whether this measurement correlates with airway smooth muscle (ASM) remodeling in asthma is unknown. In this study, we sought to determine the ability of EBUS to predict histological ASM remodeling in normal and equine asthmatic airways. We studied 109 isolated bronchi from the lungs of 13 horses. They underwent EBUS examination using a 30 MHz radial probe before being processed for histology. ASM remodeling parameters were e...
Transthecal Arthroscopy of the Palmar Distal Interphalangeal Joint in the Horse: A Cadaver Study. To describe a transthecal approach to the palmar pouch of the distal interphalangeal joint (DIPJ) in horses and compare it with the conventional blind arthroscopic technique. Methods: Ex vivo study. Methods: Cadaver forelimbs (n = 26 pairs) from mature horses. Methods: One limb from each pair was randomly assigned to each arthroscopic approach (conventional or transthecal). The conventional arthroscopic approach was performed as previously described and the transthecal approach was performed through sharp dissection of the distal reflection of the digital flexor tendon sheath (DFTS). The propo...
Association of Factor V Secretion with Protein Kinase B Signaling in Platelets from Horses with Atypical Equine Thrombasthenia. Two congenital bleeding diatheses have been identified in Thoroughbred horses: Glanzmann thrombasthenia (GT) and a second, novel diathesis associated with abnormal platelet function in response to collagen and thrombin stimulation. Objective: Platelet dysfunction in horses with this second thrombasthenia results from a secretory defect. Methods: Two affected and 6 clinically normal horses. Methods: Ex vivo study. Washed platelets were examined for (1) expression of the αIIb-β3 integrin; (2) fibrinogen binding capacity in response to ADP and thrombin; (3) secretion of dense and α-granules; (...
RADIOGRAPHIC AND ANATOMIC CHARACTERISTICS OF DORSAL HOOF WALL LAYERS IN NONLAMINITIC HORSES. 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...
Penetration depth of corneal cross-linking with riboflavin and UV-A (CXL) in horses and rabbits. CXL penetration depth is an important variable influencing clinical treatment effect and safety. The purposes of this study were to determine the penetration depth of CXL in rabbit and equine corneas in epithelium-on and epithelium-off procedures and to assess an ex vivo fluorescent biomarker staining assay for objective assessment of CXL penetration depth. Methods: CXL treatment was performed according to a standardized protocol on 21 and 17 rabbit eyes and on 12 and 10 equine eyes with and without debridement, respectively. Control corneas were treated similarly, but not exposed to CXL. Hem...
Preliminary data on the effect of osseous anatomy on ex vivo joint mobility in the equine thoracolumbar region. 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...
Evaluation of a Novel Screw Position in a Type III Distal Phalanx Fracture Model: An Ex Vivo Study. Mechanical evaluation of a novel screw position used for repair in a type III distal phalanx fracture model and assessment of solar canal penetration (SCP). Methods: Experimental study. Methods: Disarticulated equine hooves (n = 24) and 24 isolated distal phalanges. Methods: Hooves/distal phalanges cut in a sagittal plane were repaired with 1 of 2 different cortical screw placements in lag fashion. In group 1 (conventional screw placement), the screw was inserted halfway between the proximal border of the solar canal (SC) and the subchondral bone surface on a line parallel to the dorsal co...
Ex-Vivo Evaluation of a Modified Teno Fix(®) Device Repair Pattern Versus a 3-Loop Pulley for Repair of Equine Flexor Tendons. To compare the load to ultimate failure, load to a 2 mm gap, mode of failure, and gap at failure (mm) of the Teno Fix(®) (TF) to the 3-loop pulley (3LP) for repair of equine tendon lacerations. The use of 4TF devices versus 5TF devices on load to ultimate failure and load to a 2 mm gap was also compared. Methods: Ex vivo biomechanical study; unbalanced incomplete block (horse) design. Methods: Cadaveric equine forelimb superficial digital flexor tendon (n = 20 paired tendons). Methods: Ten tendon pairs were selected randomly and repaired with a 3LP and 4TF pattern, and 10 tendon pairs...
