Bone marrow-derived multipotent mesenchymal stromal cells from horses after euthanasia

Abstract Allogeneic equine multipotent mesenchymal stromal cells (eMSCs) have been proposed for use in regenerative therapies in veterinary medicine. A source of allogeneic eMSCs might be the bone marrow from euthanized horses. The purpose of this study was to compare in vitro characteristics of equine bone marrow derived eMSC (eBM-MSCs) from euthanized horses (eut-MSCs) and from narcotized horses (nar-MSCs). Eut-MSCs and nar-MSCs showed typical eMSC marker profiles (positive: CD44, CD90; negative: CD11a/CD18 and MHCII) and possessed tri-lineage differentiation characteristics. Although CD105 and MHCI expression varied, no differences were detected between eut-MSCs and nar-MSCs. Proliferation characteristics did not differ between eut-MSCs and nar-MSCs, but age dependent decrease in proliferation and increase in MHCI expression was detected. These results suggest the possible use of eut-MSCs for therapeutic applications and production of commercial available eBM-MSC products

Effect of single intralesional treatment of surgically induced equine superficial digital flexor tendon core lesions with adipose-derived mesenchymal stromal cells: a controlled experimental trial

Background: Adipose tissue is a promising source of mesenchymal stromal cells (MSCs) for the treatment of tendon disease. The goal of this study was to assess the effect of a single intralesional implantation of adipose tissue-derived mesenchymal stromal cells (AT-MSCs) on artificial lesions in equine superficial digital flexor tendons (SDFTs). Methods: During this randomized, controlled, blinded experimental study, either autologous cultured AT-MSCs suspended in autologous inactivated serum (AT-MSC-serum) or autologous inactivated serum (serum) were injected intralesionally 2 weeks after surgical creation of centrally located SDFT lesions in both forelimbs of nine horses. Healing was assessed clinically and with ultrasound (standard B-mode and ultrasound tissue characterization) at regular intervals over 24 weeks. After euthanasia of the horses the SDFTs were examined histologically, biochemically and by means of biomechanical testing. Results: AT-MSC implantation did not substantially influence clinical and ultrasonographic parameters. Histology, biochemical and biomechanical characteristics of the repair tissue did not differ significantly between treatment modalities after 24 weeks. Compared with macroscopically normal tendon tissue, the content of the mature collagen crosslink hydroxylysylpyridinoline did not differ after AT-MSC-serum treatment (p = 0.074) while it was significantly lower (p = 0.027) in lesions treated with serum alone. Stress at failure (p = 0.048) and the modulus of elasticity (p = 0.001) were significantly lower after AT-MSC-serum treatment than in normal tendon tissue. Conclusions: The effect of a single intralesional injection of cultured AT-MSCs suspended in autologous inactivated serum was not superior to treatment of surgically created SDFT lesions with autologous inactivated serum alone in a surgical model of tendinopathy over an observation period of 22 weeks. AT-MSC treatment might have a positive influence on collagen crosslinking of remodelling scar tissue. Controlled long-term studies including naturally occurring tendinopathies are necessary to verify the effects of AT-MSCs on tendon disease

Stem Cells in Equine Veterinary Practice—Current Trends, Risks, and Perspectives

With this Editorial, we introduce the Special Issue “Adipose-Derived Stem Cells and Their Extracellular Microvesicles (ExMVs) for Tissue Engineering and Regenerative Medicine Applications„ to the scientific community. In this issue, we focus on regenerative medicine, stem cells, and their clinical application

Laparoscopic resection of an exostosis of the os pubis in a horse

OBJECTIVE: To report the diagnostic findings and laparoscopic removal of an exostosis of the os pubis in a horse. STUDY DESIGN: Case report. ANIMAL: One 12-year-old Black Forest draught gelding. METHODS: History included recurrent colic before and during urination and poor performance. Findings at rectal examination included a pointed osseous prominence adjacent to the symphysis of the pecten ossis pubis. Cystoscopy revealed that this prominence caused a protrusion of the bladder wall into the lumen. Standing laparoscopy and laparoscopy under general anesthesia were performed. RESULTS: After a failed attempt at standing laparoscopy, the horse was anesthetized, and the exostosis of the os pubis was removed laparoscopically without complications. No recurrence of clinical signs associated with the exostosis was detected 12 months postoperatively. CONCLUSION: Minimally invasive surgical resection of an exostosis of the os pubis was achieved under general anesthesia with appropriately designed instruments. This treatment alleviated symptoms associated with the exostosis, including potential injury of the urinary bladder wall

Pneumoperitoneum as an uncommon complication after an axillary laceration in a horse

Lacerations of the axillary region occur frequently in horses. Typical complications caused by entrapment of air in the wound during locomotion are subcutaneous emphysema, with consecutive pneumomediastinum and pneumothorax. In this case report, the clinical, radiographic and laboratory diagnosis and management of these complications after an axillary laceration that finally resulted in pneumoperitoneum are described. A 1‐year‐old Hannoveranian was presented with a pre‐existing axillary laceration of unknown duration and subcutaneous emphysema in the surrounding tissue. Due to extensive tissue loss, attempts to adequately close the wound surgically and by packing with sterile dressing material were unsuccessful. Despite stall confinement and tying of the horse, subcutaneous emphysema was progressive and pneumomediastinum as well as pneumothorax was developed. These complications were monitored radiographically. On day 5 after admission, signs of air accumulation were detected on radiographs craniodorsally in the peritoneum and a pneumoperitoneum was diagnosed. Repeated thoracentesis with a teat cannula to gradually evacuate the thoracic cavity was used in combination with nasal oxygen insufflation to treat global respiratory insufficiency. Subcutaneous emphysema and all other complications resolved progressively and the horse was discharged from the hospital 21 days after admission when the axillary wound was adequately filled with granulation tissue. The wound healed fully 1 month later and the horse did not develop long‐term complications within the following year. To the authors´ knowledge, the development of pneumoperitoneum including its radiographic monitoring following an axillary laceration has not been described in horses previously

Effects of controlled hypoxemia or hypovolemia on global and intestinal oxygenation and perfusion in isoflurane anesthetized horses receiving an alpha-2-agonist infusion

Abstract Background Aim of this prospective experimental study was to assess effects of systemic hypoxemia and hypovolemia on global and gastrointestinal oxygenation and perfusion in anesthetized horses. Therefore, we anesthetized twelve systemically healthy warmblood horses using either xylazine or dexmedetomidine for premedication and midazolam and ketamine for induction. Anesthesia was maintained using isoflurane in oxygen with either xylazine or dexmedetomidine and horses were ventilated to normocapnia. During part A arterial oxygen saturation (SaO2) was reduced by reducing inspiratory oxygen fraction in steps of 5%. In part B hypovolemia was induced by controlled arterial exsanguination via roller pump (rate: 38 ml/kg/h). Mean arterial blood pressure (MAP), heart rate, pulmonary artery pressure, arterial and central venous blood gases and cardiac output were measured, cardiac index (CI) was calculated. Intestinal microperfusion and oxygenation were measured using laser Doppler flowmetry and white-light spectrophotometry. Surface probes were placed via median laparotomy on the stomach, jejunum and colon. Results Part A: Reduction in arterial oxygenation resulted in a sigmoid decrease in central venous oxygen partial pressure. At SaO2 < 80% no further decrease in central venous oxygen partial pressure occurred. Intestinal oxygenation remained unchanged until SaO2 of 80% and then decreased. Heart rate and pulmonary artery pressure increased significantly during hypoxemia. Part B: Progressive reduction in circulating blood volume resulted in a linear decrease in MAP and CI. Intestinal perfusion was preserved until blood loss resulted in MAP and CI lower 51 ± 5 mmHg and 40 ± 3 mL/kg/min, respectively, and then decreased rapidly. Conclusions Under isoflurane, intestinal tissue oxygenation remained at baseline when arterial oxygenation exceeded 80% and intestinal perfusion remained at baseline when MAP exceeded 51 mmHg and CI exceeded 40 mL/kg/min in this group of horses. Trial registry number 33.14-42,502-04-14/1547

Sternal bone marrow derived equine multipotent mesenchymal stromal cells (MSCs): investigations considering the sampling site and the use of different culture media

Aspiration of equine sternal bone marrow is required for the cultivation of bone marrow-derived multipotent mesenchymal stromal cells (BM-MSCs) for regenerative therapies. For bone marrow aspiration as well as for MSC cultivation, there is a need to optimize techniques and protocols to enhance MSC harvest at minimized culture times. In a comparative study bone marrow aspirates from sternebra 4 and 5 were collected at two different positions within the sternebrae, either from 10 mm or from 30 mm dorsal from the ventral margin of the sternebrae. Accuracy of the puncture depth was confirmed by ultrasonography and computed tomography. Isolated MSCs were cultivated using media supplemented with three alternative sera, i.e. fetal calf serum, standardized horse serum and autologous serum. Due to morphological characteristics (spherical shape, only thin layer of hyaline cartilage at the ventral site, reliable bone marrow aspiration from only 10 mm intraosseous depth), sternebra 5 appeared most suitable for bone marrow aspiration. Cultivation and expansion of BM-MSCs was most efficient using fetal calf serum

Effect of heel elevation on breakover phase in horses with laminitis

BACKGROUND: In a laminitic horse, the maximal loading of the toe region occurs during the breakover phase. To date, no kinetic data demonstrates the effect of supportive orthopaedic therapy in horses with laminitis on breakover phase. Thus, the purpose of this study was to examine the effect of heel elevation on the breakover phase. Eight horses with acute laminitis treated medically as well as with application of a hoof cast with heel wedge (HCHW) were included in this study. Immediately following cessation of clinical signs of acute laminitis, two measurements using the Hoof™ System were taken: the first with HCHW and the second immediately following removal of the HCHW, i.e. in barefoot condition (BFC). The hoof print was divided into three regions: toe, middle hoof, and heel. Kinetic parameters included vertical force (VF), stance duration, contact area (CA) for all hoof regions during stance phase, duration of breakover, VF in the toe region at onset of breakover and location of centre of force. RESULTS: The VF and CA were higher in the heel region (63 and 61%, respectively) and decreased significantly after removal of the HCHW (43 and 28% after removal, respectively). The breakover phase in horses with HCHW lasted 2% of stance phase and was significantly shorter than that in BFC, which lasted 6% of stance phase. The VF at onset of breakover for the toe region in horses with HCHW was significantly lower than that in BFC. The centre of the force was located at the heel region in all horses with the HCHW, and at the middle the hoof region in BFC. CONCLUSIONS: Heel elevation in horses with laminitis as examined on a concrete surface significantly shortens breakover phase and decreases the vertical force in the toe region during breakover. HCHW provides adequate support to the palmar hoof structures by increasing the contact area in the heel region and incorporating the palmar part of frog and sole into weight bearing, thus decreasing the stress on the lamellae. Hoof cast with heel elevation could be a beneficial orthopaedic supportive therapy for horses suffering from acute laminitis

Bilateral Change in Vertical Hoof Force Distribution in Horses with Unilateral Forelimb Lameness before and after Successful Diagnostic Anaesthesia

Kinetic examinations of horses with induced lameness as well as the effect of perineural anaesthesia in sound horses have shown promise, but clinical studies regarding the effect of diagnostic anaesthesia during the different stance phases are rare. Fourteen horses with unilateral forelimb lameness were examined with the Hoof™ System during trot to assess vertical force distribution (in kg) affecting both front hooves before and after diagnostic anaesthesia during landing, midstance, and breakover. For statistical analysis, a covariance analysis with repeated measurements regarding the limb (lame/sound) as well as anaesthesia (before/after) and the covariable body weight was performed. The p-values for the pairwise comparisons were adjusted using the Bonferroni–Holm correction (p p p p p = 0.001). Pressure measurements with the Hoof™ System can be used to evaluate the effect of diagnostic anaesthesia in a clinical setting with pain-related vertical force asymmetries being neutralised after diagnostic anaesthesia. Breakover is the main event influenced by lameness