Intra-articular bone marrow mononuclear cell therapy improves lameness from naturally occurring equine osteoarthritis.

Osteoarthritis (OA) can be debilitating and is related to impaired resolution of synovial inflammation. Current treatments offer temporary relief of clinical signs, but have potentially deleterious side effects. Bone marrow mononuclear cells (BMNC) are a rich source of macrophage progenitors that have the ability to reduce OA symptoms in people and inflammation in experimentally-induced synovitis in horses. The objective of this study was to evaluate the ability of intra-articular BMNC therapy to improve clinical signs of naturally occurring equine OA. Horses presenting with clinical and radiographic evidence of moderate OA in a single joint were randomly assigned to 1 of 3 treatment groups: saline (negative control), triamcinolone (positive control), or BMNC (treatment group). Lameness was evaluated subjectively and objectively, joint circumference measured, and synovial fluid collected for cytology and growth factor/cytokine quantification at 0, 7, and 21 days post-injection. Data were analyzed using General Estimating Equations with significance set at p < 0.05. There were no adverse effects noted in any treatment group. There was a significant increase in synovial fluid total nucleated cell count in the BMNC-treated group on day 7 (median 440; range 20-1920 cells/uL) compared to day 0. Mononuclear cells were the predominant cell type across treatments at all time points. Joint circumference decreased significantly in the BMNC-treated group from days 7 to 21 and was significantly lower at day 21 in the BMNC-treated group compared to the saline-treated group. Median objective lameness improved significantly in the BMNC group between days 7 and 21. GM-CSF, IL-1ra, IGF-1, and TNF-α were below detectable limits and IL-6, IL-1ß, FGF-2 were detectable in a limited number of synovial fluid samples. Inconsistent and limited differences were detected over time and between treatment groups for synovial fluid PGE2, SDF-1, MCP-1 and IL-10. Decreased lameness and joint circumference, coupled with a lack of adverse effects following BMNC treatment, support a larger clinical trial using BMNC therapy to treat OA in horses.

In vitro effects of three equimolar concentrations of methylprednisolone acetate, triamcinolone acetonide, and isoflupredone acetate on equine articular tissue cocultures in an inflammatory environment.

OBJECTIVE To compare the effects of 3 equimolar concentrations of methylprednisolone acetate (MPA), triamcinolone acetonide (TA), and isoflupredone acetate (IPA) on equine articular tissue cocultures in an inflammatory environment. SAMPLE Synovial and osteochondral explants from the femoropatellar joints of 6 equine cadavers (age, 2 to 11 years) without evidence of musculoskeletal disease. PROCEDURES From each cadaver, synovial and osteochondral explants were harvested from 1 femoropatellar joint to create cocultures. Cocultures were incubated for 96 hours with (positive control) or without (negative control) interleukin (IL)-1ß (10 ng/mL) or with IL-1ß and MPA, TA, or IPA at a concentration of 10-4, 10-7, or 10-10M. Culture medium samples were collected from each coculture after 48 and 96 hours of incubation. Concentrations of prostaglandin E2, matrix metalloproteinase-13, lactate dehydrogenase, and glycosaminoglycan were determined and compared among treatments at each time. RESULTS In general, low concentrations (10-7 and 10-10M) of MPA, TA, and IPA mitigated the inflammatory and catabolic (as determined by prostaglandin E2 and matrix metalloproteinase-13 quantification, respectively) effects of IL-1ß in cocultures to a greater extent than the high (10-4M) concentration. Mean culture medium lactate dehydrogenase concentration for the 10-4M IPA treatment was significantly greater than that for the positive control at both times, which was suggestive of cytotoxicosis. Mean culture medium glycosaminoglycan concentration did not differ significantly. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that the in vitro effects of IPA and MPA were similar to those of TA at clinically relevant concentrations (10-7 and 10-10M).

Characterization of basal and lipopolysaccharide-induced microRNA expression in equine peripheral blood mononuclear cells using Next-Generation Sequencing.

The innate immune response to lipopolysaccharide contributes substantially to the morbidity and mortality of gram-negative sepsis. Horses and humans share an exquisite sensitivity to lipopolysaccharide and thus the horse may provide valuable comparative insights into this aspect of the inflammatory response. MicroRNAs, small non-coding RNA molecules acting as post-transcriptional regulators of gene expression, have key roles in toll-like receptor signaling regulation but have not been studied in this context in horses. The central hypothesis of this study was that lipopolysaccharide induces differential microRNA expression in equine peripheral blood mononuclear cells in a manner comparable to humans. Illumina Next Generation Sequencing was used to characterize the basal microRNA transcriptome in isolated peripheral blood mononuclear cells from healthy adult horses, and to evaluate LPS-induced changes in microRNA expression in cells cultured for up to four hours. Selected expression changes were validated using quantitative reverse-transcriptase PCR. Only miR-155 was significantly upregulated by LPS, changing in parallel with supernatant tumor necrosis factor-α concentration. Eight additional microRNAs, including miR-146a and miR-146b, showed significant expression change with time in culture without a clear LPS effect. Target predictions indicated a number of potential immunity-associated targets for miR-155 in the horse, including SOCS1, TAB2 and elements of the PI3K signaling pathway, suggesting that it is likely to influence the acute inflammatory response to LPS. Gene alignment showed extensive conservation of the miR-155 precursor gene and associated promoter regions between horses and humans. The basal and LPS-stimulated microRNA expression pattern characterized here were similar to those described in human leukocytes. As well as providing a resource for further research into the roles of microRNAs in immune responses in horses, this will facilitate inter-species comparative study of the role of microRNAs in the inflammatory cascade during endotoxemia and sepsis.

Farriery for hoof wall defects: quarter cracks and toe cracks.

Conditions that result in the loss of the structural integrity of the hoof wall, such as quarter and toe cracks, are not uncommon and usually manifest in lameness. The successful management of these problems involves identifying and addressing the underlying causes, stabilization of the foot, and committed follow-up.

Managing obesity in pasture-based horses.

Obesity - a common problem in pasture-based horses - warrants intervention because it is associated with an increased risk for development of laminitis. Treating obesity in pasture-based horses is relatively simple and generally involves reducing caloric intake by using grazing muzzles and/or increasing caloric expenditure through exercise. To prevent recurrence of obesity after weight loss, clients should be educated on how to monitor body condition and to adjust feeding and management programs to maintain proper body condition.

Clinical assessment of nutritional status and feeding programs in horses.

Veterinarians are a primary source of nutritional information and advice for horse owners. This article reviews methods for clinical assessment of nutritional status and feeding programs that can be applied to an individual horse or group of horses. Physical examination, including measurement of body weight and evaluation of body condition score, estimation of nutrient requirements and the nutrient content of the horse's diet, and evaluation of the feeding method are important components of the assessment. Ongoing clinical assessment of health and body condition will gauge the need for reassessment of the feeding plan. Obvious indications for prompt reevaluation of diet and feeding include changes in health status (eg, body condition), life stage or physiologic state (eg, pregnancy), or performance status.

Vascular distribution of contrast medium during intraosseous regional perfusion of the distal portion of the equine forelimb.

OBJECTIVE: To describe the vascular distribution pattern of contrast medium during intraosseous regional perfusion (IORP) of the distal portion of the equine forelimb. SAMPLE POPULATION: 13 cadaveric forelimbs from 12 horses without forelimb diseases. PROCEDURES: Serial lateromedial radiographic views were taken of the distal portion of 10 heparinized cadaveric forelimbs at 0, 1, 2, 6, 15, and 30 minutes during IORP of the third metacarpal bone (MCIII) by use of iodinated contrast medium and a tourniquet placed over the proximal portion of MCIII. Vascular regions of interest (ROI) were created for each radiograph. Reviewers identified the presence or absence of contrast medium-induced opacified vessels in all ROI on radiographs. This information was summarized to identify vessel-filling patterns over time. Vessel identification was verified by use of computed tomography angiography and latex perfusion studies on the distal portion of separate cadaveric forelimbs. RESULTS: During IORP, contrast medium filled the medullary cavity of the MCIII; exited via transcortical vessels; and diffused distally to the remaining arteries and veins of the forelimb, distal to the tourniquet. Maximum vessel and soft tissue opacification occurred in most specimens at 6 and 30 minutes, respectively. Serial radiography vessel patterns matched those of computed tomography images and dissected specimens. CONCLUSIONS AND CLINICAL RELEVANCE: IORP provides a repeatable pattern of vascular distribution in the distal portion of the equine forelimb. To our knowledge, our study provides the first documentation of arterial perfusion by use of IORP; results of previous reports indicate that IORP delivers medications to only the venous vessels of the perfused forelimb.

Hand-assisted laparoscopic ovariohysterectomy in the mare.

OBJECTIVE: To develop a minimally invasive, hand-assisted laparoscopic ovariohysterectomy (HALS-OHE) technique in the mare and to evaluate safety and any associated complications. STUDY DESIGN: Experimental study. ANIMALS: Eight, 11-24-year-old mares with anatomically normal urogenital tracts. METHODS: The surgical technique was developed in 2 non-survival mares and subsequently evaluated in 6 survival procedures. Food was withheld for 48 hours, then mares were anesthetized and positioned in dorsal recumbency for laparoscopic surgery. A hand access device (Omniport) was placed followed by 4 laparoscopic portals. Transection of the ovarian pedicles and broad ligaments was achieved using a combination of a laparoscopic stapling instrument (Endo GIA II), an ultrasonically activated instrument (Harmonic Scalpel), and endoscopic clips (Endo Clip II ML). The genital tract was exteriorized through the laparotomy, and the uterus transected and sutured in a conventional pattern. Horses were evaluated through postoperative day 14 when a necropsy was performed. RESULTS: Four mares recuperated well after surgery, 1 mare was euthanatized because of bilateral femur fracture during anesthetic recovery, and another developed severe pleuropneumonia. At necropsy all but 1 abdominal incision was healing routinely. One mare had abscessed along the celiotomy incision and developed visceral adhesions. Uncomplicated healing of transected mesovarial, mesometrial, and uterine remnants was observed. CONCLUSIONS: Ovariohysterectomy in horses can be accomplished using HALS technique. CLINICAL RELEVANCE: HALS-OHE technique represents a minimally invasive and technically feasible alternative for conventional OHE. Careful patient selection and preparation may reduce the complications observed. The HALS technique may be useful in other laparoscopic surgical procedures.

Elution of metronidazole and gentamicin from polymethylmethacrylate beads.

OBJECTIVE: To characterize the elution and bioactivity of metronidazole and gentamicin sulfate polymerized, individually and in combination, with polymethylmethacrylate (PMMA). STUDY DESIGN: In vitro experimental study. METHODS: PMMA beads containing metronidazole (3 concentrations), gentamicin sulfate, or metronidazole and gentamicin sulfate were immersed in 5 mL of phosphate-buffered saline in triplicate. Eluent was replaced at specified time intervals for 1 or 21 days, and antibiotic concentrations were measured by high-performance liquid chromatography. Changes in antibiotic bioactivity attributable to polymerization or copolymerization of the antibiotics with PMMA, ethylene oxide sterilization, and storage of AIPMMA beads containing metronidazole were evaluated. RESULTS: Antibiotic elution patterns were similar for all groups. Day 1 elution for groups containing metronidazole or gentamicin individually represented a mean 63%-66% and 79%, respectively, of the 21-day total. Approximately 50% of the day 1 elution occurred during the first hour. The elution of metronidazole was dose dependent. The elution of metronidazole (day 3-21) and gentamicin (all days) was significantly greater when metronidazole and gentamicin were combined (P <.05). The addition of metronidazole delayed polymerization of PMMA. Neither polymerization nor copolymerization of metronidazole and gentamicin with PMMA, gas sterilization, or 2-month storage of beads containing metronidazole significantly affected antimicrobial bioactivity. CONCLUSIONS: Metronidazole elution from PMMA was dose dependent. Copolymerization of metronidazole and gentamicin sulfate in PMMA resulted in increased rates of elution. Intraoperative preparation of metronidazole-impregnated PMMA beads is not practical, but sterilization and storage for 2 months should not affect efficacy. CLINICAL RELEVANCE: The local delivery of biologically active metronidazole and gentamicin by elution from PMMA is feasible.

Evaluation of the harmonic scalpel for laparoscopic bilateral ovariectomy in standing horses.

OBJECTIVE: To evaluate use of the Harmonic Scalpel (Ethicon Endo-Surgery Inc., Cincinnati, OH) for performing laparoscopic bilateral ovariectomy in standing horses. STUDY DESIGN: Experimental study. ANIMALS: Eight mares aged 2-20 years and weighing 410-540 kg. METHODS: Standing laparoscopic bilateral ovariectomy was performed in 8 mares with normal reproductive tract anatomy. The Harmonic Scalpel (an ultrasonically activated instrument) was used to simultaneously transect and obtain hemostasis of the ovarian pedicle. Necropsy was performed on 4 mares 3 days after surgery and 4 mares 30 days after surgery. Gross and histopathologic evaluation of the ovarian pedicles was performed to characterize tissue reaction. RESULTS: Complete hemostasis of the ovarian pedicles was obtained in all mares. Median transection time for the ovarian pedicle was 28 minutes. Postoperative complications included transient fever, moderate subcutaneous emphysema, and incisional seroma formation. On necropsy examination, there were no signs of generalized peritonitis, postoperative hemorrhage, or adhesion formation. Mild to moderate acute inflammation and scar formation with moderate chronic inflammation at the ovarian pedicle was found at 3 and 30 days. Median depth of coagulation necrosis at 3 days was 2.87 mm. CONCLUSIONS: The Harmonic Scalpel appears to provide reliable hemostasis of the ovarian pedicle during elective laparoscopic ovariectomy in horses. Clinical Relevance-The Harmonic Scalpel represents a safe alternative to other methods of hemostasis during elective laparoscopic ovariectomy in horses.

Experimental evaluation of urinary bladder marsupialization in male goats.

OBJECTIVE: To evaluate the outcome of urinary bladder marsupialization in male goats. STUDY DESIGN: Prospective, experimental study. ANIMALS: Six healthy mixed-breed male goats. METHODS: After experimentally induced urethral obstruction, 6 male goats had urinary bladder marsupialization. Renal ultrasonography, complete blood count, and serum biochemical analysis were evaluated preoperatively (day 0), at 7 postoperative days, and then at 30-day intervals until 180 days. Stomal diameter was recorded immediately postoperatively and at each postoperative interval. Necropsy examination was performed on day 180 or when stomal stricture or death occurred. RESULTS: Stomal stricture occurred in 1 goat at 120 days. Another goat was found dead at 150 days; severe, suppurative cystitis was identified on necropsy. All goats had mild urine scald dermatitis. Serum biochemical values remained within normal limits, but significant decreases in white blood cell count, serum creatinine concentration, and stomal diameter occurred. At necropsy, all bladders were tubular in shape. Histological evidence of chronic suppurative cystitis and chronic, mild lymphoplasmacytic pyelitis occurred in all goats. Bacterial culture of renal tissue yielded growth in 3 goats, and bladder mucosal swabs yielded bacterial growth in all goats. CONCLUSIONS: Although clinical signs of ascending urinary tract infection were not observed in goats with patent stomata, urinary bladder marsupialization may result in ascending urinary tract inflammation or infection. CLINICAL RELEVANCE: Based on our results, urinary bladder marsupialization should be recommended with caution as the primary method for management of urinary tract obstruction in clinical cases.