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.
Aimonetti E, Genton M, Lischer CJ, Rossignol F.To determine the benefit of using a three-dimensional (3D)-printed titanium aiming device for placing two 5.5-mm transarticular screws during proximal interphalangeal joint (PIPJ) arthrodesis in equine species. Methods: Ex vivo study. Methods: A total of 40 cadaveric forelimbs from adult horses. Methods: A senior surgeon and a first-year resident each performed 20 constructs: 10 using the device and 10 using a freehand technique. Procedure duration and number of intraoperative radiographs were recorded. Final radiographs were evaluated by an ECVS-certified surgeon unaware of treatment group, b...
Márquez-Moya A, Carreras-Vico N, Sala-Ayala L, Martínez-Boví R, Cuervo-Arango J.Transvaginal ultrasound-guided follicular aspiration or ovum pick-up (OPU) has become the standard technique for oocyte collection in mares for intracytoplasmic sperm injection (ICSI). Although repeated follicular flushes and wall scraping are commonly used to improve oocyte recovery rate (ORR), the relative contribution of each remains unclear. This study aimed to compare the effects of multiple flushes versus controlled scraping time on ORR in mares. A controlled trial was conducted in two phases: (1) an ex vivo phase using slaughterhouse ovaries (n = 32), and (2) an in vivo phase in c...
Sullivan J, Blea J, McKemie DS, Kass PH, Knych HK.The pharmacokinetics and pharmacodynamics of betamethasone following intra-articular administration to horses have been described; however, studies characterizing intramuscular administration are lacking. Twenty-four horses received an intramuscular dose of 12 mg betamethasone sodium phosphate/betamethasone acetate. Blood and urine were collected at post administration for up to 408 h. Concentrations of betamethasone were determined using LC-MS/MS and pharmacokinetic parameters determined using a Population PK three-compartment model. The duration of pharmacodynamic effects was assessed by...
Hoerdemann M, Sahoo DK, Allbaugh RA, Kubai MA.To assess if an inexpensive, commercially available ultraviolet C (UV-C) light device with a peak emission of 275 nm can inhibit equine keratomycosis-associated pathogens located at different corneal depths in an ex vivo model. Methods: A controlled, randomized experimental design. Aspergillus fumigatus and Fusarium solani isolates were inoculated in fresh bovine corneoscleral transplants, superficially or at 450 μm of stromal depth for the ulcerative keratomycosis or stromal abscess model, respectively. After a minimum of 18 h of incubation, treatment groups received 15 s of UV-C li...
Profaska M, Zarzycka M, Dubniewicz K, Witkowski M, Wieczorek J, Gil D, Wafula S, Lanh DK, Kotula-Balak M.Mare endometrosis remains a poorly understood pathological process. Objective: Mare endometrial tissue with endometrosis was used to determine the effects of relaxin (RLX). Healthy tissues that were left untreated (H), or treated with vehicle (Hveh), and tissues with endometrosis that were left untreated (E), treated with vehicle (Eveh), and treated with RLX (10, 25, 50 nM; ER) were used for an ex vivo system for 72 h. Methods: Tissue histological examination, and immunoenzymatic measurement of the concentrations of transforming growth factor β (TGF-β), interleukins (IL-6 and IL-8), and pr...
Sullivan J, Blea J, Morales CJ, McKemie DS, Kass PH, Knych HK.Corticosteroids, such as isoflupredone, are effective anti-inflammatory medications and as such are commonly used to treat inflammation associated with training and injuries in performance horses. While the pharmacokinetics and pharmacodynamics of isoflupredone acetate (IPA) following intra-articular administration to horses has been well described, studies characterizing intramuscular (IM) administration are lacking. The objective of the current study was to describe the pharmacokinetics and anti-inflammatory effects of IPA following IM administration to horses. Twelve horses received a singl...
Lepage H, de Chaisemartin C, Spadaro Rosselo A, Leroy H, Lepage O.To describe and report clinical outcomes after transcutaneous guttural pouch (GP) catheterization (TGPC) in standing horses. Methods: Ex vivo study and case series. Methods: One cadaver head, records of 10 normal horses and 14 horses treated with TGPC. Methods: Relevant anatomical landmarks were determined through dissection of one cadaveric specimen and 10 normal radiographic studies. Records of horses diagnosed with empyema or mycosis and treated with standing TGPC were reviewed for complications that occurred during or after the operation. Results: Ex vivo and radiological studies identifie...
Mitlyng N, Hobbs KJ, Cooper BL, Sheats MK.Lipopolysaccharide (LPS) is frequently used in equine research to model clinical endotoxemia; however, there is no standardized protocol for inducing cytokine production in equine whole blood. To address this knowledge gap, the goal of this study was to compare the cytokine responses elicited by three different LPS stimulation protocols. Whole blood was collected from six healthy horses (aged 5-30 years; mixed breeds and genders) from the North Carolina State University teaching herd (IACUC #23-412). Sixty milliliters of heparinized blood were aseptically drawn and divided into 15 mL aliquots....
Buckman PN, Brinker BA, Kapeller LE, Komáromy AM.To determine the accuracy of the EYEMATE-SC suprachoroidal tracer for telemetric tonometry in canine and equine globes. Methods: The EYEMATE-SC sensor (7.8 mm × 3.8 mm × 1 mm) was implanted in the suprachoroidal space of four freshly enucleated normal canine and two normal equine eyes. The anterior chambers were cannulated and connected to a reservoir of Plasma-Lyte A and a manometer. Starting at a manometric IOP of 5 mmHg, the pressure was progressively increased to 80 mmHg by raising the reservoir. At each setpoint (5, 10, 15, 20, 25, 30, 35, 40, 50, 60, 70, and 80 mmHg), t...
Ysebaert MP, Johnson J, Marie U, Campos A, Verchrerin A, Ducharme NG, Rossignol F, Luedke LK.To compare the biomechanical stability of a novel arytenoid implant (tie-bolt, TB) with a suture anchor (SA) and standard laryngoplasty using suture alone (control) in equine larynges. Methods: Randomized ex vivo controlled experimental study. Methods: Thirty equine cadaver larynges. Methods: Larynges were randomly assigned to TB, SA, or control groups (n = 10 each). The force of abduction was measured for each construct, then constructs underwent cyclic loading (5-50 N for 3000 cycles) to assess elongation, followed by load-to-failure testing to assess ultimate load, displacement, stiffn...
Bosch K, Zsoldos RR, Hartig A, Licka T.The influence of soft tissue structures, including ligaments spanning one or more intervertebral junctions and the nuchal ligament, on motion of the equine cervical joints remains unclear. The present study addressed this using four post-mortem horse specimens extending from head to withers with all ligaments intact. Three-dimensional kinematics was obtained from markers on the head and bone-anchored markers on each cervical and the first thoracic vertebra during rotation, lateral bending, flexion and extension of the whole head, and neck segment. Yaw, pitch, and roll angles in 8 cervical join...
Beggan CP, Panizzi L, Oliver LJ.To measure microsphere recovery following needle-through-and-through lavage (NTAT) of the metacarpo-/metatarsophalangeal joint (fetlock) and digital flexor tendon sheath (DFTS) compared to endoscopic lavage (EL). Methods: Ex vivo experimental study. Methods: Adult equine cadavers immediately following euthanasia (n = 10). Methods: Colored 15 μm microspheres (2 million) were injected into fetlock joints and DFTS. Synovial structures were assigned to NTAT or EL groups. Each lavage was performed using 5 L of 0.9% NaCl, sequentially collecting egress fluid for microsphere quantification. Re...
Baldwin CM, Gillen A.To evaluate three different jejunal transection angles for end-to-end jejunojejunostomies, comparing construction time, lumen size, and suture number. Methods: Ex vivo cadaver study. Methods: Eight euthanized horses each had three mid-jejunal specimens harvested. Methods: Jejunal segments were assigned to one of three groups (A30, A45, and A60) and transected at 30, 45, or 60°, respectively. Following transection, a standardized handsewn single-layer interrupted modified Lembert anastomosis was performed, and construction time and number of sutures placed were recorded. Anastomotic index (AI)...
Kadic LIM, Liu CC, Leise BS, McCauley CT, Riggs LM.The aim of the study was to determine if a novel arthrodesis technique for the equine metacarpophalangeal (MCP) joint using a human non-contact bridging (NCB), polyaxial locking compression plate system has biomechanical and potential clinical advantages versus a traditional 5.5 mm stainless steel locking compression plate (LCP) under biomechanical testing. Methods: An ex vivo biomechanical equine cadaver study. Methods: A total of 11 MCP joints collected from adult Thoroughbred horses euthanized unrelated to musculoskeletal disease. Methods: Each pair of MCP joints were randomly implanted w...
Kodama S, Maeda T, Mita H, Tamura N, Chiba K, Matsukawa M.Despite the prevalence of lower limb disease in racehorses, outdoor bone evaluation equipment is lacking. Consequently, a secure and cost-effective field screening system for bones is required to identify early-stage leg illness. Elastic stiffness cannot be measured using x-ray techniques and is closely associated with bone quality, which is recognized as a significant determinant of fracture risk. Quantitative ultrasound has garnered attention as a viable option, offering insights into elastic stiffness. Understanding the characteristic behaviors of ultrasonic wave properties (velocity, atten...
Tress D, Hennessy S, Merle R, Jensen KC, Lischer C, Ehrle A.Impingement of spinous processes (SPs) is commonly diagnosed in the equine athlete. For diagnostic and therapeutic purposes, local injections are performed at the level of the space between adjacent spinous processes in affected horses. Objective: To assess the accuracy of different techniques for the local injection of the interspinal space in the equine thoracolumbar spine. Methods: Ex vivo experimental study. Methods: Equine thoracolumbar spine specimens were used to compare three techniques for needle insertion (midline; bilateral abaxial; unilateral oblique), two needles (20G-1½″; 20G-...
Gao T, Liu X, Qiu D, Li Y, Qiu Z, Qi J, Li S, Guo X, Zhang Y, Wang Z, Gao X, Ma Y, Ma T.Cefquinome is used to treat septicemia caused by () and respiratory infections caused by subsp. in foals. However, studies reporting the use of cefquinome to target as pathogens of sepsis are lacking. Therefore, this study aimed to determine the optimal dosage regimen for cefquinome against using a PK/PD model. After the administration of 1 mg/kg cefquinome (intramuscularly or intravenously), blood samples were collected at different time points to determine the serum concentration of cefquinome via HPLC. The pharmacokinetic parameters were evaluated via NCA (WinNonlin 5.2.1 software). Th...
Sheahan BJ, Schubert AG, Schubert W, Sheats MK, Schnabel LV, Gilbertie JM.Neutrophil extracellular traps (NETs) play a significant role in response to a variety of infectious and inflammatory stimuli in human and veterinary medicine. Although entrapment of bacteria can be an important function of NETs, the exuberant release of DNA and other intracellular molecules has also been negatively implicated in the pathogenesis of different diseases. Thus, NET formation must be tightly controlled and represents an opportunity for therapeutic interventions. Horses are particularly sensitive to bacterial stimuli that have previously been shown to cause NETs in other species, b...
Li D, Wang X, Yao J, Chen S.Adipose tissue-derived extracellular vesicles (ATEVs) have garnered attention for their roles in intercellular communication and regulation. This study presents the first investigation of equine adipose tissue, with ATEV being extracted concurrently during the isolation of adipose-derived mesenchymal stem cells (ADSCs). Through CCK-8 cell proliferation assays and scratch migration assays, a significant promotional effect of ATEV on ADSCs was observed, which not only accelerated the proliferation rate of the stem cells but also enhanced their migratory capacity. The application of Trypan Blue e...
Knych HK.Pharmacokinetics is the study of the movement of drug in the body and includes the processes of absorption, distribution, metabolism, and excretion. Pharmacodynamics is the pharmacologic effect of the drug on the body. The pharmacokinetics of a drug determines its pharmacologic effect. Pharmacokinetic studies describe drug concentrations while pharmacodynamics allow for assessment of drug effects. Combined pharmacokinetic/pharmacodynamic studies allow for integration of drug concentrations with pharmacologic effect. Data generated from pharmacokinetic studies can be especially useful in establ...
CT arthrography (CTA) for evaluating cartilage defects would be beneficial in clinical practice. Since the contrast medium (CM) volume is often driven by operator experience, the aim was to evaluate the minimum volume of iodinated CM for CTA sufficient to identify iatrogenic cartilage defects in horse fetlock. The study was conducted on ex-vivo 32 distal limbs from adult horses collected at slaughter. Iatrogenic cartilage defects were arthroscopically made on fetlock, at 5 predetermined locations. Subsequently, CT scans were performed, according to the following protocol: precontrast scan, fol...
Nugent Z, Jensen A, Owen N, Peffers AJ, Moothanchery M, Peffers MJ.Equine odontoclastic tooth resorption and hypercementosis (EOTRH) is a painful disorder primarily affecting the incisor teeth of horses over 15 years of age. Clinical signs of the disease include prehension problems, halitosis and in severe cases weight loss. The disease predominately affects the reserve crown and presents as a loss of dental tissue and excessive build-up of cementum. Objective: To determine the radiographic scores of horses with EOTRH and age-matched controls and to increase understanding of EOTRH using microCT to compare teeth from horses with EOTRH and age-matched control...
Korsós SA, Kibleur P, Josipovic I, Boone M, Vlaminck L.In orthograde endodontic treatments, different methods are available to debride the pulp canals of endodontically compromised equine cheek teeth, but their efficacy is unknown. Objective: To explore and compare the efficacy and anatomical changes caused by manual versus reciprocating filing techniques in equine cheek teeth, to explore the presence of instrumentation mishaps described in human dentistry and to explore anatomical complexities of the pulp cavity that often remain uninstrumented using microcomputed tomography (μCT). Methods: Ex-vivo randomised experiments. Methods: Twenty-two ext...
Verhaar N, Hammer E, Reineking W, Hewicker-Trautwein M, Geburek F.To compare the practicability and tissue sample quality between different intestinal biopsy techniques. Methods: Experimental, randomized ex vivo study. Methods: Small intestine of nine horses. Methods: Four different biopsy techniques were evaluated in the aboral jejunum and the ileum within 1 h after euthanasia. One segment was used as control (C), and the applied techniques included an 8 mm biopsy punch (BP), transverse wedge resection (TW), longitudinal wedge resection with transverse closure (LW) and a longitudinal sample using Eppendorfer biopsy forceps (EF). Defects were closed using ...
Vaughn SA, Berghaus LJ, Hart KA.Breed differences exist between horses and ponies in circulating concentrations of several hormones, notably ACTH and insulin. These hormones regulate stress and metabolic responses, but in other species, they also impact leukocyte oxidant responses. The effects of these hormones on equine leukocytes have not been evaluated to date. If equine leukocytes are similarly regulated, breed differences in increased plasma hormone concentrations or altered sensitivity to them at the leukocyte level could result in breed-related differences in oxidant responses or oxidative status. The objective of thi...
St Blanc MP, Mirza MH, Riggs LM, Leise BS.The aim of the study was to evaluate time to closure, bursting pressures and luminal diameters of a single and double-layer end-to-end anastomoses in normal equine descending colon. Methods: Experimental study. Methods: Eight adult horses. Methods: Four segments of descending colon from each horse were randomly assigned to a control (n = 8, CON), one-layer (n = 12, group 1; continuous Lembert pattern), or two-layer group (n = 12, group 2; simple continuous oversewn with Cushing pattern). Anastomoses were performed, and time to closure and luminal diameter were measured. Bursting pr...
Irandoust S, O'Neil LM, Stevenson CM, Franseen FM, Ramzan PHL, Powell SE, Brounts SH, Loeber SJ, Ergun DL, Whitton RC, Henak CR, Muir P.Catastrophic injury has a low incidence but leads to the death of many Thoroughbred racehorses. Objective: To determine sensitivity, specificity, and reliability for third metacarpal condylar stress fracture risk assessment from digital radiographs (DR) and standing computed tomography (sCT). Methods: Controlled ex vivo experiment. Methods: A blinded set of metacarpophalangeal joint DR and sCT images were prepared from 31 Thoroughbreds. Four observers evaluated the condyles and parasagittal grooves (PSG) of the third metacarpal bone for the extent of dense bone and lucency/fissure and assigned...
Woodrow JS, Palmisano M, Kulp J, Hopster K.Complications from general anesthesia, including pneumonia and decreased wound healing, are influenced by changes in immune cell function secondary to sedatives and anesthetics. It was hypothesized that immune cell function would be depressed in the early postanesthetic period. The objective was to investigate airway immune cell function before and after a general anesthetic episode in an equine in vivo model using ex vivo cell stimulations with lipopolysaccharide (LPS) for assessment of immune function. Methods: Prospective experimental study. Methods: Six healthy, adult, institution-owned ...
Portaels J, Van Crombrugge E, Van Den Broeck W, Lagrou K, Laval K, Nauwynck H.Numerous Aspergillus fumigatus (Af) airborne spores are inhaled daily by humans and animals due to their ubiquitous presence. The interaction between the spores and the respiratory epithelium, as well as its impact on the epithelial barrier function, remains largely unknown. The epithelial barrier protects the respiratory epithelium against viral infections. However, it can be compromised by environmental contaminants such as pollen, thereby increasing susceptibility to respiratory viral infections, including alphaherpesvirus equine herpesvirus type 1 (EHV-1). To determine whether Af spores di...
Baudisch N, Singer E, Jensen KC, Eichler F, Meyer HJ, Lischer C, Ehrle A.Surgical treatment options for horses with overriding dorsal spinous processes include interspinous ligament desmotomy and partial spinous process ostectomy. The impact of spinal surgery on the three-dimensional biomechanics of the equine thoracolumbar spine and the epaxial musculature is unclear. Objective: To investigate the influence of interspinous ligament desmotomy and cranial wedge ostectomy on the biomechanics of the equine thoracolumbar spine and the paraspinal Musculi multifidi. Methods: Ex-vivo experiments. Methods: Twelve equine thoracolumbar spine specimens were mounted in a custo...
Toupadakis CA, Wong A, Genetos DC, Cheung WK, Borjesson DL, Ferraro GL, Galuppo LD, Leach JK, Owens SD, Yellowley CE.To determine the optimal osteogenic source of equine mesenchymal stem cells (eMSCs) and optimize collection of and expansion conditions for those cells. Methods: 10 adult Quarter Horses and 8 newborn Thoroughbred foals. Methods: eMSCs were isolated from bone marrow (BM), adipose tissue, and umbilical cord blood and tissue, and the osteogenic potential of each type was assessed. Effects of anatomic site, aspiration volume, and serum type on eMSC yield from BM were investigated. Results: BM-eMSCs had the highest overall expression of the osteogenic genes Cbfa1, Osx, and Omd and staining for ALP ...
Gilbertie JM, Schnabel LV, Hickok NJ, Jacob ME, Conlon BP, Shapiro IM, Parvizi J, Schaer TP.Bacterial invasion of synovial joints, as in infectious or septic arthritis, can be difficult to treat in both veterinary and human clinical practice. Biofilms, in the form of free-floating clumps or aggregates, are involved with the pathogenesis of infectious arthritis and periprosthetic joint infection (PJI). Infection of a joint containing an orthopedic implant can additionally complicate these infections due to the presence of adherent biofilms. Because of these biofilm phenotypes, bacteria within these infected joints show increased antimicrobial tolerance even at high antibiotic concentr...
Côté O, Clark ME, Viel L, Labbé G, Seah SY, Khan MA, Douda DN, Palaniyar N, Bienzle D.Secretoglobin family 1A member 1 (SCGB 1A1) is a small protein mainly secreted by mucosal epithelial cells of the lungs and uterus. SCGB 1A1, also known as club (Clara) cell secretory protein, represents a major constituent of airway surface fluid. The protein has anti-inflammatory properties, and its concentration is reduced in equine recurrent airway obstruction (RAO) and human asthma. RAO is characterized by reversible airway obstruction, bronchoconstriction and neutrophilic inflammation. Direct effects of SCGB 1A1 on neutrophil functions are unknown. We have recently identified that the SC...
Goodrich LR, Choi VW, Carbone BA, McIlwraith CW, Samulski RJ.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,...
Lavoie-Lamoureux A, Moran K, Beauchamp G, Mauel S, Steinbach F, Lefebvre-Lavoie J, Martin JG, Lavoie JP.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...
Anderson JR, Phelan MM, Foddy L, Clegg PD, Peffers MJ.Osteoarthritis is an age-related degenerative musculoskeletal disease characterized by loss of articular cartilage, synovitis, and subchondral bone sclerosis. Osteoarthritis pathogenesis is yet to be fully elucidated with no osteoarthritis-specific biomarkers in clinical use. equine cartilage explants ( = 5) were incubated in tumor necrosis factor-α (TNF-α)/interleukin-1β (IL-1β)-supplemented culture media for 8 days, with the media removed and replaced at 2, 5, and 8 days. Acetonitrile metabolite extractions of 8 day cartilage explants and media samples at all time points underwent one-d...
Wang XF, Lin YZ, Li Q, Liu Q, Zhao WW, Du C, Chen J, Wang X, Zhou JH.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...
Veraa S, Bergmann W, van den Belt AJ, Wijnberg I, Back W.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...
Bullone M, Beauchamp G, Godbout M, Martin JG, Lavoie JP.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...
Van Cleemput J, Poelaert KCK, Laval K, Impens F, Van den Broeck W, Gevaert K, Nauwynck HJ.Pollens are well-known triggers of respiratory allergies and asthma. The pollen burden in today's ambient air is constantly increasing due to rising climate change and air pollution. How pollens interact with the respiratory mucosa remains largely unknown due to a lack of representative model systems. We here demonstrate how pollen proteases of Kentucky bluegrass, white birch and hazel selectively destroy integrity and anchorage of columnar respiratory epithelial cells, but not of basal cells, in both ex vivo respiratory mucosal explants and in vitro primary equine respiratory epithelial cells...
Vietmeier J, Niedorf F, Bäumer W, Martin C, Deegen E, Ohnesorge B, Kietzmann M.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 ...
Hiller LP, Stover SM, Gibson VA, Gibeling JC, Prater CS, Hazelwood SJ, Yeh OC, Martin RB.An important concept in bone mechanics is that osteons influence mechanical properties in several ways, including contributing to toughness and fatigue strength by debonding from the interstitial matrix so as to "bridge" developing cracks. Observations of "pulled out" osteons on fracture surfaces are thought to be indicative of such behavior. We tested the hypothesis that osteon pullout varies with mode of loading (fatigue vs. monotonic), cortical region, elastic modulus, and fatigue life. Mid-diaphseal beams from the dorsal, medial, and lateral regions of the equine third metacarpal bone were...
Mouser VHM, Dautzenberg NMM, Levato R, van Rijen MHP, Dhert WJA, Malda J, Gawlitta D.The implantation of chondrocyte-laden hydrogels is a promising cartilage repair strategy. Chondrocytes can be spatially positioned in hydrogels and thus in defects, while current clinical cell therapies introduce chondrocytes in the defect depth. The main aim of this study was to evaluate the effect of spatial chondrocyte distribution on the reparative process. To reduce animal experiments, an ex vivo osteochondral plug model was used and evaluated. The role of the delivered and endogenous cells in the repair process was investigated. Full thickness cartilage defects were created in equine ost...
Naumann S, Meyer JP, Kiesow A, Mrestani Y, Wohlrab J, Neubert RH.The penetration behavior into human nails and animal hoof membranes of a novel antifungal agent (EV-086K) for the treatment of onychomycosis was investigated in this study. The new drug provides a high lipophilicity which is adverse for penetration into nails. Therefore, four different formulations were developed, with particular focus on a colloidal carrier system (CCS) due to its penetration enhancing properties. On the one hand, ex vivo penetration experiments on human nails were performed. Afterwards the human nail plates were cut by cryomicrotome in order to quantify the drug concentratio...
Radunsky D, Blumenfeld-Katzir T, Volovyk O, Tal A, Barazany D, Tsarfaty G, Ben-Eliezer N.Multi-echo spin-echo (MESE) protocol is the most effective tool for mapping T relaxation in vivo. Still, MESE extensive use of radiofrequency pulses causes magnetization transfer (MT)-related bias of the water signal, instigated by the presence of macromolecules (MMP). Here, we analyze the effects of MT on MESE signal, alongside their impact on quantitative T measurements. Study used 3 models: in vitro urea phantom, ex vivo horse brain, and in vivo human brain. MT ratio (MTR) was measured between single-SE and MESE protocols under different scan settings including varying echo train lengths, n...
te Moller NC, Brommer H, Liukkonen J, Virén T, Timonen M, Puhakka PH, Jurvelin JS, van Weeren PR, Töyräs J.Arthroscopy enables direct inspection of the articular surface, but provides no information on deeper cartilage layers. Optical coherence tomography (OCT), based on measurement of reflection and backscattering of light, is a diagnostic technique used in cardiovascular surgery and ophthalmology. It provides cross-sectional images at resolutions comparable to that of low-power microscopy. The aim of this study was to determine if OCT is feasible for advanced clinical assessment of lesions in equine articular cartilage during diagnostic arthroscopy. Diagnostic arthroscopy of 36 metacarpophalangea...
Drobnic M, Radosavljevic D, Cör A, Brittberg M, Strazar K.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...
Seeto WJ, Tian Y, Winter RL, Caldwell FJ, Wooldridge AA, Lipke EA.A common challenge in cell therapy is the inability to routinely maintain survival and localization of injected therapeutic cells. Delivering cells by direct injection increases the flexibility of clinical applications, but may cause low cell viability and retention rates due to the high shear forces in the needle and mechanical wash out. In this study, we encapsulated endothelial colony forming cells (ECFCs) in poly(ethylene glycol)-fibrinogen (PF) hydrogel microspheres using a custom-built microfluidic device; this system supports rapid encapsulation of high cell concentrations (10 million c...
Vaccaro C, Busetto R, Bernardini D, Anselmi C, Zotti A.To evaluate the precision and accuracy of assessing bone mineral density (BMD) by use of mean gray value (MGV) on digitalized and digital images of conventional and digital radiographs, respectively, of ex vivo bovine and equine bone specimens in relation to the gold-standard technique of dual-energy x-ray absorptiometry (DEXA). Methods: Left and right metatarsal bones from 11 beef cattle and right femurs from 2 horses. Methods: Bovine specimens were imaged by use of conventional radiography, whereas equine specimens were imaged by use of computed radiography (digital radiography). Each specim...
Virén T, Huang YP, Saarakkala S, Pulkkinen H, Tiitu V, Linjama A, Kiviranta I, Lammi MJ, Brünott A, Brommer H, Van Weeren R, Brama PA, Zheng YP....The aim of this study was to compare sensitivity of ultrasound and optical coherence tomography (OCT) techniques for the evaluation of the integrity of spontaneously repaired horse cartilage. Articular surfaces of horse intercarpal joints, featuring both intact tissue and spontaneously healed chondral or osteochondral defects, were imaged ex vivo with arthroscopic ultrasound and laboratory OCT devices. Quantitative ultrasound (integrated reflection coefficient (IRC), apparent integrated backscattering coefficient (AIB) and ultrasound roughness index (URI)) and optical parameters (optical refle...
Tidd N, Michelsen J, Hilbert B, Quinn JC.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...
Baghi HB, Nauwynck HJ.The mucosal surfaces are important sites of entry for a majority of pathogens, and viruses in particular. The migration of antigen presenting cells (APCs) from the apical side of the mucosal epithelium to the lymph node is a key event in the development of mucosal immunity during viral infections. However, the mechanism by which viruses utilize the transmigration of these cells to invade the mucosa is largely unexplored. Here, we establish an ex vivo explant model of monocytic cell transmigration across the nasal mucosal epithelium and lamina propria. Equine nasal mucosal CD172a(+) cells (nmCD...
Van Cleemput J, Poelaert KCK, Laval K, Nauwynck HJ.Equine herpesvirus type 5 (EHV5) is a ubiquitous, yet obscure pathogen in the horse population and is commonly associated with fatal equine multinodular pulmonary fibrosis (EMPF). To date, little is known about the precise pathogenesis of EHV5. Here, we evaluated the dynamics of EHV5 infection in representative ex vivo and in vitro equine models, using immunofluorescence staining and virus titration. EHV5 was unable to infect epithelial cells lining the mucosa of nasal and tracheal explants. Similarly, primary equine respiratory epithelial cells (EREC) were not susceptible to EHV5 following in...
Busoni V, Heimann M, Trenteseaux J, Snaps F, Dondelinger RF.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...
Gallhoefer NS, Spiess BM, Guscetti F, Hilbe M, Hartnack S, Hafezi F, Pot SA.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...
Shibeshi W, Abraham G, Kneuer C, Ellenberger C, Seeger J, Schoon HA, Ungemach FR.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...
Yaneselli K, Barrachina L, Remacha AR, Algorta A, Vitoria A, Cequier A, Romero A, Vázquez FJ, Maisonnave J, Rodellar C.Propagation ex vivo of mesenchymal stem cells (MSCs) requires culture medium supplementation. Fetal bovine serum (FBS) has long been the gold standard supplement, but its use is being questioned mainly due to ethical and safety issues. The use of platelet lysate (PL) as substitute of FBS has been proposed but little is known about its effects on equine MSCs characteristics including their immune profile. The aim of this work was to investigate for the first time the effect of allogenic PL on the immunogenic and immunomodulatory gene expression profile of equine bone marrow derived MSCs (eBM-MS...
Schnabel CL, Steinig P, Schuberth HJ, Koy M, Wagner B, Wittig B, Juhls C, Willenbrock S, Murua Escobar H, Jaehnig P, Feige K, Cavalleri JM.Leukocytes and their functional capacities are used extensively as biomarkers in immunological research. Commonly employed indicators concerning leukocytes are as follows: number, composition in blood, response to discrete stimuli, cytokine release, and morphometric characteristics. In order to employ leukocytes as biomarkers for disease and therapeutic monitoring, physiological variations and influencing factors on the parameters measured have to be considered. The aim of this report was to describe the ranges of selected leukocyte parameters in a sample of healthy horses and to analyse wheth...
Hamza E, Gerber V, Steinbach F, Marti E.Horses are particularly prone to allergic and autoimmune diseases, but little information about equine regulatory T cells (Treg) is currently available. The aim of this study therefore was to investigate the existence of CD4(+) Treg cells in horses, determine their suppressive function as well as their mechanism of action. Freshly isolated peripheral blood mononuclear cells (PBMC) from healthy horses were examined for CD4, CD25 and forkhead box P3 (FoxP3) expression. We show that equine FoxP3 is expressed constitutively by a population of CD4(+) CD25(+) T cells, mainly in the CD4(+) CD25(high)...
Hinrichs K.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 ...
