Andrew J. Dart

Superficial digital flexor tendonitis in the horse

The superficial digital flexor tendon (SDFT) is an elastic structure that during maximal exercise appears to operate close to its functional limits. The biomechanical and biochemical responses to exercise, injury, and healing are still poorly understood but ongoing research is providing valuable new information which is addressed in this review. It appears that the SDFT matures early, after which time it has limited ability to adapt to stress and undergoes progressive degeneration. Focal hypocellularity, collagen fibril degeneration, selective fibril loading and alterations in the noncollagenous matrix occur primarily within the central core region of the midmetacarpal segment. Current treatment strategies have had equivocal results in returning animals to optimal athletic activity. To date it would seem that progressive rehabilitation programmes coupled with regular ultrasonographic evaluations are a cost-effective and comparable strategy when compared to surgical treatment methods. Recent interest in pharmacological modulation of intrinsic healing of collagenous structures has led to the investigation of various growth factors as potential therapeutic aids in the healing of tendon injuries. However, one of the major goals in tendon research, and one which holds the most optimism for success in the immediate future, is the prevention of tendon injuries.

Modulation of aggrecan and ADAMTS expression in ovine tendinopathy induced by altered strain

OBJECTIVE To evaluate histologic, immunohistochemical, and molecular changes in tendon induced by altered strain in a large-animal model. METHODS A full-thickness partial-width laceration of the infraspinatus tendon was created in 5 sheep, while 5 sham-operated sheep were used as controls. Sheep were killed after 4 weeks, and 4 differentially stressed tendon regions (tensile or near bone attachment from overstressed or stress-deprived halves) were evaluated for histopathology, proteoglycan (PG) accumulation, and characterization of glycosaminoglycans and aggrecan catabolites. Gene expression of matrix components, enzymes, and inhibitors was analyzed by reverse transcriptase-polymerase chain reaction. RESULTS Histopathologic changes were detected in both overstressed and stress-deprived tensile tendon, but only in stress-deprived tendon near bone. In overstressed and stress-deprived tensile tendon, levels of keratan sulfate, chondroitin 4-sulfate, and chondroitin 6-sulfate were increased. In overstressed tensile tendon, levels of ADAMTS-generated aggrecan catabolites were increased. There was increased matrix metalloproteinase 13 (MMP-13) and decreased fibromodulin and decorin expression in all regions. Increased MMP-1, MMP-9, MMP-14, and ADAMTS-1 expression, and decreased type II collagen expression were restricted to stress-deprived tendon. In stress-deprived bone-attachment regions, messenger RNA (mRNA) for aggrecan was decreased, and ADAMTS was increased. In overstressed tensile tendon, aggrecan mRNA was increased, and ADAMTS was decreased. CONCLUSION The distinct molecular changes in adjacent tissue implicate altered strain rather than humoral factors in controlling abnormal tenocyte metabolism, and highlight the importance of regional sampling. Tendon abnormalities induced by increased strain are accompanied by increased aggrecan, decreased ADAMTS, and low PG expression, which may negatively impact the structural integrity of the tissue and predispose to rupture.

Mechanical Destabilization Induced by Controlled Annular Incision of the Intervertebral Disc Dysregulates Metalloproteinase Expression and Induces Disc Degeneration

Study Design. An investigation of mechanical destabilization of the lumbar ovine intervertebral disc (IVD) inducing IVD degeneration (IVDD) as determined by multiparameter outcome measures (magnetic resonance imaging [MRI], IVD composition, biomechanical testing, gene profiling, immunohistochemistry, and immunoblotting). Objective. To assess the effect of IVD mechanical destabilization on matrix protein and metalloproteinase gene expression to investigate the pathophysiological mechanisms of lumbar IVDD. Summary of Background Data. Several earlier studies have used annular transection to induce IVDD in sheep, but none have optimized or validated the most appropriate lesion size. Methods. The annulus fibrosus (AF) incision inducing maximal change in IVD biomechanics was applied to L1–L2, L3–L4, and L5–L6 discs in vivo to compare with a sham procedure at 3 months post operation. IVDs were evaluated by MRI, biomechanics, histopathology, proteoglycan and collagen content, gene expression, and aggrecan proteolysis by Western blotting. Results. Significant changes were observed in lesion (6 × 20 mm2) compared with sham IVDs at 3 months post operation: reduced disc height on MRI; increased neutral zone in biomechanical testing; depleted proteoglycan and collagen content in the nucleus pulposus (NP) and lesion half of the AF but not in the contralateral AF; increased messenger RNA for collagen I and II, aggrecan, versican, perlecan, matrix metalloproteinase (MMP)-1 & 13, and ADAMTS-5, in the lesion-site AF and NP but not in the contralateral AF. ADAMTS-4 messenger RNA was increased in the lesion-site AF but decreased in the NP. Despite an upregulation in MMPs, there was no change in MMP- or ADAMTS-generated aggrecan neoepitopes in any region of the IVD in lesion or sham discs. Conclusion. Lumbar IVDD was reproducibly induced with a 6 × 20 mm2 annular lesion, with focal dysregulation of MMP gene expression, cell cloning in the inner AF, loss of NP aggrecan, and disc height. Loss of aggrecan from the NP was not attributable to increased proteolysis in the interglobular domain by MMPs or ADAMTS.

Effect of bandaging on second intention healing of wounds of the distal limb in horses.

OBJECTIVE To evaluate the effect of a non-occlusive dressing incorporated in a 3-layer bandage on second intention healing of wounds of the distal portion of the limb. STUDY DESIGN Untreated wounds in 33 adult horses used in four studies using the same wound-healing model conducted over 5 years. METHODS Standardised, full-thickness wounds were made in the skin overlying the dorsomedial aspect of the mid-metacarpus; 17 horses were bandaged with a non-occlusive dressing covered by gauze-coated cotton wool that was compressed with adhesive tape; 16 horses were left unbandaged. Wounds were photographed weekly for 9 weeks and the images were analysed electronically. RESULTS There were significant effects associated with bandage (P < 0.0001), week (P < 0.001), and bandage by week interaction (P < 0.0001). There was no difference in wound area at the first time-point after wound creation (P = 0.38). After week 1, there was a difference between bandaged and unbandaged wounds in wound area at each measurement until the end of the study. Bandaged wounds showed greater and more prolonged retraction. Unbandaged wounds retracted for 2 weeks before beginning to contract, whereas bandaged wounds continued to retract for 3 weeks. In bandaged wounds excess granulation tissue required regular trimming, but not in unbandaged wounds. There was no difference between groups in the total days to healing or the overall rate of healing. CONCLUSIONS These results should be treated with caution until validated with contemporaneous, controlled studies. Covering a wound with a non-occlusive dressing in a 3-layer bandage led to greater wound retraction, modulated the rate of wound contraction and promoted excessive granulation tissue. If excessive granulation tissue is excised regularly, bandaging has no effect on total time to healing.

A Preliminary Study on the Effect of Manuka Honey on Second‐Intention Healing of Contaminated Wounds on the Distal Aspect of the Forelimbs of Horses

OBJECTIVE To determine the effect of manuka honey on second-intention healing of contaminated, full-thickness skin wounds in horses. STUDY DESIGN Experimental. ANIMALS Adult Standardbred horses (n = 8). METHODS One wound was created on the dorsomedial aspect of the third metacarpus in both forelimbs, contaminated with feces, and bandaged for 24 hours. Bandages were removed and wounds rinsed with isotonic saline solution. Wounds on 1 limb had manuka honey applied daily (n = 8) whereas wounds on the contralateral limb received no treatment (n = 8). Bandages were replaced and changed daily for 12 days, after which treatment stopped, bandages were removed, leaving wounds open to heal. Wound area was measured 24 hours after wound creation (day 1), then weekly for 8 weeks. Overall time for healing was recorded. Wound area and rate of healing of treated and control wounds were compared statistically. RESULTS Treatment with manuka honey decreased wound retraction and treated wounds remained significantly smaller than control wounds until day 42; however, there was no difference in overall healing time between treatment and control wounds. CONCLUSIONS Treatment with manuka honey reduced wound area by reducing retraction but did not affect overall healing time of full-thickness distal limb wounds using this wound-healing model.

Recombinant equine growth hormone administration: effects on synovial fluid biomarkers and cartilage metabolism in horses

REASONS FOR PERFORMING STUDY Recombinant equine growth hormone (reGH) has recently been evaluated for effects on body condition and wound healing. It has the potential to influence articular cartilage via stimulation of IGF-1. OBJECTIVES To investigate effects of administration on synovial joint metabolism. METHODS Six mature horses were given 20 microg/kg bwt reGH daily for 8 weeks by i.m. injection. Three control horses were injected with sterile water. Serum and synovial fluid samples were collected at 6, 8, 11 and 16 weeks for GH and IGF-1 assays. Articular cartilage harvested at week 16 was evaluated by Western analysis using monoclonal antibodies BC-13, BC-4, 8-A-4 and CH-3. RESULTS Concentrations of IGF-1 in serum and synovial fluid were significantly elevated (P < 0.05) at 6 and 8 weeks in the reGH group. Glycosaminoglycan concentrations in synovial fluid were significantly less than controls at these time points, suggesting that reGH may modulate proteoglycan metabolism in articular cartilage. In the reGH group, there were not any alterations in synovial fluid content of 3B3(-) epitope or aggrecan metabolite, or in aggrecan or link protein catabolites retained within cartilage, that might be expected with development of osteoarthritis. CONCLUSIONS Intramuscular administration of reGH may be a more efficient means of delivery of IGF-1 to joints for cartilage resurfacing initiatives. POTENTIAL RELEVANCE We found no alterations in cartilage metabolism indicative of development of osteoarthritis.

Effect of Prosthesis Number and Position on Rima Glottidis Area in Equine Laryngeal Specimens

OBJECTIVE To assess the effect of 1 or 2 laryngeal prosthetic sutures on rima glottidis areas in equine laryngeal specimens. STUDY DESIGN Experimental, randomized design. ANIMALS Cadaveric equine larynges (n=16). METHODS Larynges were collected from 10 horses; 2 sutures each were preplaced in the right and left sides of each larynx. A dorsal suture (DS) was placed through the caudal rim of the dorsal midline of the cricoid cartilage, under the cricopharyngeus muscle and through the proximal and rostral aspect of the muscular process. A lateral suture (LS) was placed 1.5 cm lateral to the DS and through the muscular process more distal and caudal to the 1st suture. Larynges were positioned in a customized stand and the rima glottidis photographed after each suture (LS or DS) or suture combination (CS) was tied in random sequence. An additional 6 larynxes were used to determine whether the tension applied to the sutures was repeatable. Sutures were preplaced in both the right and left side of each larynx as described above and each suture and CS was tied and released 3 times in each larynx. Photographs were taken of the rima glottidis after each suture or CS was tied generating 3 replicates for each suture configuration on each side of the 6 larynges. RESULTS Mean rima glottidis area was not different between DS and LS when tied alone (P=.85); however, mean area after CS (DS+LS) was greater than DS (P<.001) and LS (P<.001) alone. The coefficient of variation for the 6 suture patterns were low (1-7%) and the intraclass correlation coefficient estimates were very high (0.997-0.998) demonstrating excellent repeatability between replicates for each of the 3 suture configurations. CONCLUSION Our results suggest that laryngoplasty using 2 prostheses; 1 placed dorsally in the cricoid and through the rostral and proximal muscular process and 1 placed 1.5 cm lateral to the 1st and more caudal and distal in the muscular process results in a greater cross sectional area of the rima glottidis than either suture used alone. CLINICAL RELEVANCE Seemingly prosthetic sutures contribute independently to each other in determining the contour of the rima glottidis. Use of 2 prosthetic sutures improves crosssectional area of the rima glottidis compared with each suture alone and may improve surgical outcome in laryngoplasty.

Surgical resection of a dysgerminoma in a mare.

A mare was referred for further evaluation of a mass found in the left caudal abdomen during a routine postpartum reproductive palpation. The mare was clinically normal with no history of health problems. Ultrasonographic examination of the mass confirmed its presence, but the origin of the mass could not be accurately determined. Routine haematology and serum biochemistry results were within normal limits. The mare was initially treated conservatively with antibiotics, but the mass continued to increase in size, so it was surgically excised. The mass involved the left ovary. The mare showed transient abdominal pain after surgery, but developed no other complications and was in foal 7 months later. On histology, the mass was diagnosed as a dysgerminoma, a rare ovarian tumour of germ cell origin.

Retrospective study of 103 presumed cases of tick (Ixodes holocyclus) envenomation in the horse

OBJECTIVE   Review 103 cases of presumed tick envenomation in horses. DESIGN   Retrospective study. METHOD   Variables, including date of presentation, age, breed, weight, presence of ticks, gait and respiration scores, duration of recumbency, treatment, outcome and complications were recorded. A series of univariable screening tests were performed and used in a multivariable logistic regression model. RESULTS   There were a total of 103 cases affecting 10 breeds, aged between 1 week and 18 years of age. Horses >6 months old and weighing >100 kg had a higher odds of death than those <6 months old and <100 kg. Cases were seen from North Queensland to the central coast of New South Wales and were more likely to present in the warmer months. There was no association between the number of ticks found on an animal and death. Horses with a higher respiratory score had higher odds of dying, but there was no association between gait score and survival. Horses recumbent >120 h after presentation had higher odds of dying. Complications were reported in 35% of horses. The odds ratio for survival was higher for horses receiving >0.5 mL/kg of tick antiserum. Overall, 74% of horses survived. Multivariable modelling was limited by the small sample size. CONCLUSION   In general, tick envenomation in horses follows the geographic distribution of Ixodes holocyclus. Tick antiserum administered at >0.5 mL/kg increases the odds of survival. It would appear that the complications associated with managing a recumbent horse increase the odds of death.

Effects of intra‐articular sodium pentosan polysulfate and glucosamine on the cytology, total protein concentration and viscosity of synovial fluid in horses

OBJECTIVE Evaluate synovial fluid cytology, total protein concentration and viscosity after intra-articular administration of combined pentosan polysulfate and glucosamine in horses. DESIGN Ten adult Standardbred horses had each carpal joint (n = 20) randomly assigned to one of two treatments: control (5 mL saline) or treated (pentosan polysulfate and glucosamine). All horses received an intra-articular injection every 7 days for 3 weeks and synovial fluid samples were collected on days 1 (baseline), 2, 3, 7, 8, 9, 14, 15, 16 and 21. Synovial fluid variables measured included total protein concentration, total nucleated cell count, red blood cell count, the neutrophil, lymphocyte, mononuclear cell and eosinophil percentages, and viscosity. RESULTS Following injection, the total nucleated cell count increased in the synovial fluid samples from both groups. Values were significantly higher in treated joints on days 2, 3, 15 and 16. Red blood cell counts were low and no differences were seen between treated and control joints. Total protein concentrations and neutrophil percentages increased in both groups after injection, but decreased towards normal ranges within 7 days. Total protein concentration was significantly higher in treated joints on days 1, 3, 8, 9 and 15. Percentages of neutrophils were higher in treated joints on day 2, but significantly lower on days 8 and 15. Viscosity was significantly higher in treated joints on day 2 only. CONCLUSIONS Intra-articular administration of combined pentosan polysulfate and glucosamine causes a mild inflammatory synovitis that is not substantially different to that elicited by injection of a similar volume of saline and so we conclude that these drugs are safe to use in the horse.