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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 Autophagy Marker LC3 Is Processed during the Sperm Capacitation and the Acrosome Reaction and Translocates to the Acrosome Where It Colocalizes with the Acrosomal Membranes in Horse Spermatozoa. 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...
Comparison of Two Bandage Splint Constructs in an Ex Vivo Equine Metacarpal Fracture Model. 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, ...
Assessing the functional properties of tenogenic primed mesenchymal stem cells in ex vivo equine tendon and ligament explants: A preliminary study. 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...
Trichostatin A-Mediated Epigenetic Modulation Predominantly Triggers Transcriptomic Alterations in the Ex Vivo Expanded Equine Chondrocytes. 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...
Ex vivo and in vivo evaluation of a modified interrupted Lembert pattern for small intestinal anastomoses in horses. 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...
Ex-vivo articular cartilage removal from equine proximal interphalangeal joints using cannulated drill bits. 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...
Cannabinoid receptors are expressed in equine synovium and upregulated with synovitis. 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...
Differential expression and methylation patterns of NFATC1, NADSYN1 and JAK3 gene in equine chondrocytes expanded in monolayer culture. 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...
The effect of repeated freezing and thawing on the suture pull-out strength in equine arytenoid and cricoid cartilages. To assess the effect of repeated freezing and thawing on the suture pull-out strength in arytenoid and cricoid cartilages subjected to the laryngoplasty (LP) procedure. Methods: Ex vivo experimental study. Methods: Ten grossly normal equine cadaveric larynges. Methods: Bilateral LP constructs were created using a standard LP technique. One hemilarynx was randomly allocated to the single freeze and thaw group and the other allocated to the repeated freeze and thaw (3 complete cycles) group. The suture ends of each LP construct were attached to a load frame and subjected to monotonic loading unt...
Mechanisms of bicarbonate secretion in the equine colon ex vivo. To examine bicarbonate (HCO3-) secretion ex vivo in the equine large colon to determine any differences between the right dorsal colon (RDC) and right ventral colon (RVC). The effect of phenylbutazone (PBZ) on HCO3- secretion was examined in the RDC. Methods: 14 healthy horses. Methods: In anesthetized horses (n = 10), segments of mucosa from RDC and RVC were harvested to measure HCO3- secretion ex vivo with the pH Stat method. The effect of PBZ on HCO3- secretion in the RDC was studied in 4 additional horses. Results: Three distinct mechanisms of HCO3- secretion previously described in a muri...
Pharmacokinetics of grapiprant and effects on TNF-alpha concentrations following oral administration to horses. Grapiprant is a prostaglandin E2 receptor antagonist that has been found to be an effective anti-inflammatory in dogs and that is devoid of some of the adverse effects associated with traditional NSAIDs that elicit their effects through inhibition of PGE2 production. Previously published reports have described the pharmacokinetics of this drug in horses when administered at 2 mg/kg; however, pharmacodynamic effects in this species have yet to be described. The objective of the current study was to describe the pharmacokinetics and pharmacodynamics of grapiprant at a higher dose. Eight horses...
Influence of unilateral and bilateral vocal cordectomy on airflow across cadaveric equine larynges at different Rakestraw grades of arytenoid abduction. To assess the effect of vocal cordectomy on airflow across equine larynges at different Rakestraw grades of arytenoid abduction using a unidirectional airflow model. Methods: Ex vivo, repeated measures. Methods: Twenty cadaveric equine larynges. Methods: The right arytenoid cartilage was maximally abducted in all larynges. Each larynx was assigned a Rakestraw grade A or B, and the left arytenoid was abducted accordingly. Each larynx was tested under 3 conditions: intact, left vocal cordectomy (LVC), and bilateral vocal cordectomy (BVC). Translaryngeal pressure and airflow were measured, and di...
Effect of single and double hemitenotomy on equine deep digital flexor tendon length and strength in experimental load challenges. To evaluate a double hemitenotomy (DHT) technique as an alternative to complete deep digital flexor (DDFT) tenotomy. Methods: Experimental ex vivo study. Methods: Isolated DDFTs (n = 30) and cadaveric forelimbs (n = 16). Methods: In part 1, 15 isolated DDFT pairs were used. Two hemitenotomies were created in 1 DDFT while the other served as reference. Monotonic tensile load was applied. Tendon lengthening, load reduction, and load at failure were recorded. In part 2, 16 cadaveric forelimb pairs were subjected to DHT followed by complete tenotomy (CT) under monotonic compressive load. Diffe...
Morphological analysis of third metacarpus cartilage and subchondral bone in Thoroughbred racehorses: An ex vivo study. Racehorses are exposed to repetitive overload during training and competition, causing joint hyperextension, tissue fatigue, and ultimately skeletal failure. Some degree of bone changes, such as sclerosis, are expected in equine athletes, as adaptation to the biomechanical rigors of training and racing. Understanding the imaging characteristics of the equine joint surface and subchondral bone would allow earlier detection of injuries or adaptation, improving prognosis and training programs. This study sought to describe the joint surface structural patterns and the periarticular structures of ...
Dual-contrast micro-CT enables cartilage lesion detection and tissue condition evaluation ex vivo. Post-traumatic osteoarthritis is a frequent joint disease in the horse. Currently, equine medicine lacks effective methods to diagnose the severity of chondral defects after an injury. Objective: To investigate the capability of dual-contrast-enhanced computed tomography (dual-CECT) for detection of chondral lesions and evaluation of the severity of articular cartilage degeneration in the equine carpus ex vivo. Methods: Pre-clinical experimental study. Methods: In nine Shetland ponies, blunt and sharp grooves were randomly created (in vivo) in the cartilage of radiocarpal and middle carpal joi...
Influence of screw head diameter on ex vivo fixation of equine lateral condylar fractures with 5.5 mm cortical screws. To determine the influence of screw head diameter on equine condylar fracture fixation with 5.5 mm cortical screws. Methods: Ex vivo, biomechanical study, blinded, matched-pair design. Methods: Fifteen pairs of equine third metacarpal (MC3) bones. Methods: Lateral condylar fractures were simulated by parasagittal osteotomies and repaired pairwise by 2 × 5.5 mm cortical screws of 8 mm (standard) or 10 mm (modified) head diameter. Interfragmentary compression at maximum screw insertion torque was measured. The instrumented specimens were pairwise stratified for biomechanical testing u...
Ex vivo comparison of 3 Tesla magnetic resonance imaging and multidetector computed tomography arthrography to identify artificial soft tissue lesions in equine stifles. To determine the diagnostic performance of computed tomographic arthrography (CTA) and 3 Tesla magnetic resonance imaging (MRI) for detecting artificial meniscal, meniscotibial ligament (MTL) lesions and cruciate ligament (CL) lesions in horses. Methods: Ex vivo controlled laboratory study. Methods: Nineteen stifles from adult horses. Methods: Stablike defects (n = 84) (16 mm long, 10 mm deep) were created in the menisci (n = 35), CLs (n = 24), and MTLs (n = 25) via arthroscopy prior to MRI and CTA (80 mL contrast at 85 mg/mL per joint). Two radiologists, unaware of the lesions...
Evaluation of an ex vivo model of the blood-perfused equine uterus. Uterine pathologies are the most common causes of infertility in mares. This study aimed to establish an ex vivo blood-perfused model for equine uteri and investigate the possible effects of different cycle stages (estrus, diestrus and anestrus) on the applicability of the model. Uteri (n = 13) were collected at an abattoir, flushed with preservation solution, transported to the laboratory on ice, and isolated perfused with autologous blood for 6 h (n = 12). For negative control, one uterus was handled as described but left without perfusion for 6 h. The cycle stage was determined by ex...
Differentiation of boldenone administration from ex vivo transformation in the urine of castrated male horses. Boldenone is an anabolic-androgenic steroid that is prohibited in equine sports. However, in certain situations, it is endogenous or is believed to be formed by microbes in urine, and therefore, an approach for the differentiation is required. Following the identification of Δ1-progesterone and 20(S)-hydroxy-Δ1-progesterone as potential biomarkers of microbial activity, the presence of six steroids was investigated in the postrace urine of castrated male horses (geldings, n = 158). In line with endogenous findings from several other species when ultrasensitive methods are employed, bolde...
Effective Penetration of a Liposomal Formulation of Bleomycin through Ex-Vivo Skin Explants from Two Different Species. Bleomycin is a chemotherapy agent that, when administered systemically, can cause severe pulmonary toxicity. Bleosome is a novel formulation of bleomycin encapsulated in ultra-deformable (UD) liposomes that may be applicable as a topical chemotherapy for diseases such as non-melanoma skin cancer. To date, the ability of Bleosome to effectively penetrate through the skin has not been evaluated. In this study, we investigated the ability of Bleosome to penetrate through ex vivo skin explants from dogs and horses. We visualized the penetration of UD liposomes through the skin by transmission elec...
Ex vivo ultrasonographic and histological morphometry of small intestinal wall layers in horses. Ultrasonographic morphometry of wall layers is commonly used in veterinary patients with suspected small intestinal disease, however published studies comparing this method with histopathology in horses are limited. This prospective, methods comparison study compared the qualitative and quantitative characteristics of small intestinal wall layers using ex vivo high-frequency ultrasound versus histopathology in a sample of 16 horses. Transverse section images of duodenum, distal jejunum, and ileum were acquired with a high-frequency linear transducer (7-15 MHz). Transverse histological cryosec...
Ex vivo biomechanical evaluation of polyester and polyblend suture techniques to perform equine laryngoplasty. To validate the use of a polyblend tape suture in equine laryngoplasty (PL). Methods: Experimental study. Methods: Thirty-two cadaveric larynges. Methods: Each larynx was randomly assigned to 1 of 4 groups: PL with polyblend tape suture (TigerTape), without (TT) or with a cannula (TTC) in the muscular process of the arytenoid cartilage, and PL with polyester suture (Ethibond), without (EB) or with a cannula (EBC). Construct stiffness, total migration, creep, and drift values were measured after 3000 cycles. The specimens were then loaded to failure to assess their residual properties: load a...
Therapeutic effects of equine amniotic membrane suspension on corneal re-epithelialization and haze in a modified lagomorph ex vivo wound healing model. To investigate the therapeutic effects of topical equine amniotic membrane (eAM) suspension following corneal wounding in a controlled experimental setting. Methods: Equine amniotic membrane was collected, gamma irradiated, homogenized for topical suspension preparation, and cryopreserved. Corneoscleral rims harvested from fresh rabbit globes were wounded via keratectomy and were maintained in an air-liquid interface ex vivo corneal culture model. Treatment groups included topical gamma irradiated eAM suspension (n = 20) and a control group (n = 20). Re-epithelialization of the wound was a...
Gross, computed tomographic, and endoscopic anatomy of the equine carpal extensor tendon sheaths. (1) To describe the computed tomography (CT) and gross anatomy of the equine extensor carpi radialis sheath (ECRS) and common digital extensor sheath (CDETS); (2) to describe a single-portal endoscopic examination of the ECRS and CDETS. Methods: Ex vivo experimental. Methods: Thirty clinically normal cadaver thoracic equine limbs severed at the humeral diaphysis. Methods: Ten limbs underwent plain and intrathecal contrast CT examinations and gross dissection of the ECRS and CDETS. Single-portal endoscopic examination of ECRS and CDETS was attempted in 4 limbs and endoscopic examination was per...
Diffusion of dye after perineural injection of the palmar/plantar nerves in two different sites in horses: an ex-vivo study. This study aimed to evaluate the distribution of dye at two perineural injection sites of the palmar/plantar nerves and describe their anatomical variations of positioning at the level of the distal metacarpus/metatarsus in horses. Two ml of 1% methylene blue solution was injected in 24 limbs (12 forelimbs and 12 hindlimbs) of horse cadavers at two different points: Group 1: between the branch of the suspensory ligament and deep digital flexor tendon, and Group 2: between the deep and superficial digital flexor tendons. This was followed by dissection and evaluation of the solution diffusion. ...
A threshold volume of 10 ml is suggested for detecting articular cartilage defects in equine carpal joints using CT arthrography: Ex vivo pilot study. Computed tomographic arthrography (CTA) has been described as a method for detecting articular cartilage defects in equine carpal joints; however, published studies on the effects of contrast volume for lesion detection are currently lacking. The purpose of this prospective, experimental, pilot study was to determine a threshold volume of iodinated contrast for CTA of the antebrachiocarpal (ABC) and middle carpal (MC) joints for detection of articular cartilage surface defects. Articular cartilage defects were iatrogenically created in the surfaces of the ABC and MC joints of 20 equine cadaver...
Interleukin-6 and lactate dehydrogenase expression in a novel ex vivo rocking model of equine corneal epithelial wound healing. To establish a physiologically relevant ex vivo model of equine corneal epithelial wound healing. Methods: Fourteen equine corneas were randomly assigned to one of two groups: wounded (n = 8) or unwounded (n = 6) controls. In the wounded group, the axial corneal epithelium was removed by applying a 6 mm filter paper disk soaked in 1N-NaOH for 60 s. Corneas were subsequently cultured using an air-liquid interface model. Evaluation of corneal healing was performed daily, and culture medium was collected. Corneas were randomly assigned to undergo processing via histopathology and RNAscope i...
Fracture Resistance of Equine Cheek Teeth With and Without Occlusal Fissures: A Standardized ex vivo Model. fracture models are frequently used in human dentistry to provide insights in the fracture mechanisms of teeth. Equine cheek teeth fractures are an important dental pathology, but there has been no research performed to examine the fracture resistance . To evaluate the fracture resistance of equine cheek teeth and identify anatomical predictors that might influence fracture resistance in healthy teeth. It was further evaluated if the presence of a fissure caused a decrease in fracture resistance. experimental design. Individual cheek teeth were subjected to a compression load in a univer...
The Effect of Reducing the Bone to Cast Distance in an Equine Transfixation Pin Cast: An Ex Vivo Biomechanical Study. The aim of this study was to evaluate the effect of reducing the bone to cast distance on the resistance of the pin to cyclic loading in equine transfixation pin casts. Methods: Eleven pairs of cadaveric equine third metacarpal bones were prepared and one 6.3/8.0 mm transfixation pin was placed in standard fashion 10 mm proximal to the distal physeal scar into each bone. One metacarpus of each pair was tested with a distance of 10 mm (10 mm group) and the contralateral metacarpus with a distance of 20 mm (20 mm group) between the outer cortex of the bone and the fixation of t...
Effect of sugar metabolite methylglyoxal on equine lamellar explants: An ex vivo model of laminitis. Laminitis is one of the most devastating diseases in equine medicine, and although several etiopathogenetic mechanisms have been proposed, few clear answers have been identified to date. Several lines of evidence point towards its underlying pathology as being metabolism-related. In the carbonyl stress pathway, sugars are converted to methylglyoxal (MG)-a highly reactive α-oxoaldehyde, mainly derived during glycolysis in eukaryotic cells from the triose phosphates: D-glyceraldehyde-3-phosphate and dihydroxyacetone phosphate. One common hypothesis is that MG could be synthesized during the dig...
