Effects of tendon injury on uninjured regional tendons in the distal limb: An in-vivo study using an ovine tendinopathy model.

Following injury to a tendon little is known about potential for pathology to develop in other regional tendons from overloading or altered function. The aim of this study was to investigate the gene expression and histopathological changes that occur 1) within the deep digital flexor tendon (DDFT) after injury to the superficial digital flexor tendon (SDFT) and 2) within the flexor tendons (SDFT and DDFT) after injury to the extensor tendons. Merino wethers [Ovis aries] (n = 18) were divided into three equal groups and underwent either partial transection of the SDFT, complete transection of the extensor tendons or were left as non-operated controls. Tendons were harvested and sampled regionally for gene expression (real time PCR) and histologic analysis eight weeks after surgery. Transection of the SDFT resulted in increased expression of collagen III, versican, biglycan, lumican and MMP1 (P<0.026 for all genes) within the DDFT. There was no effect of transecting the extensor tendons on the expression of any gene tested in either the SDFT or the DDFT. The DDFT had elevated histopathology scores induced by transection of the SDFT, eight weeks previously. There were minimal histological differences in either the SDFT or DDFT after transection of the extensor tendons. Transection of the SDFT results in a mild, subclinical tendinopathy within the DDFT with potential implications on treatment and rehabilitation of SDFT injuries. Injury to the extensor tendons has minimal measured effect on the SDFT or DDFT.

Focal experimental injury leads to widespread gene expression and histologic changes in equine flexor tendons.

It is not known how extensively a localised flexor tendon injury affects the entire tendon. This study examined the extent of and relationship between histopathologic and gene expression changes in equine superficial digital flexor tendon after a surgical injury. One forelimb tendon was hemi-transected in six horses, and in three other horses, one tendon underwent a sham operation. After euthanasia at six weeks, transected and control (sham and non-operated contralateral) tendons were regionally sampled (medial and lateral halves each divided into six 3 cm regions) for histologic (scoring and immunohistochemistry) and gene expression (real time PCR) analysis of extracellular matrix changes. The histopathology score was significantly higher in transected tendons compared to control tendons in all regions except for the most distal (P ≤ 0.03) with no differences between overstressed (medial) and stress-deprived (lateral) tendon halves. Proteoglycan scores were increased by transection in all but the most proximal region (P < 0.02), with increased immunostaining for aggrecan, biglycan and versican. After correcting for location within the tendon, gene expression for aggrecan, versican, biglycan, lumican, collagen types I, II and III, MMP14 and TIMP1 was increased in transected tendons compared with control tendons (P < 0.02) and decreased for ADAMTS4, MMP3 and TIMP3 (P < 0.001). Aggrecan, biglycan, fibromodulin, and collagen types I and III expression positively correlated with all histopathology scores (P < 0.001), whereas lumican, ADAMTS4 and MMP14 expression positively correlated only with collagen fiber malalignment (P < 0.001). In summary, histologic and associated gene expression changes were significant and widespread six weeks after injury to the equine SDFT, suggesting rapid and active development of tendinopathy throughout the entire length of the tendon. These extensive changes distant to the focal injury may contribute to poor functional outcomes and re-injury in clinical cases. Our data suggest that successful treatments of focal injuries will need to address pathology in the entire tendon, and that better methods to monitor the development and resolution of tendinopathy are required.

Treatment of experimentally induced osteoarthritis in horses using an intravenous combination of sodium pentosan polysulfate, N-acetyl glucosamine, and sodium hyaluronan.

OBJECTIVE: To assess the effects of sodium pentosan polysulfate (PPS), N-acetyl glucosamine (NAG), and sodium hyaluronan (HA) in horses with induced osteoarthritis (OA). STUDY DESIGN: Experimental. ANIMALS: Adult Standard bred horses (n = 16). METHODS: OA was induced arthroscopically in 1 intercarpal joint; 8 horses were administered 3 mg/kg PPS, 4.8 mg/kg NAG, and 0.12 mg/kg HA (PGH), intravenously (IV), weekly and 8 horses were administered an equivalent volume of saline IV until study completion (day 70). Horses underwent a standardized treadmill exercise program. Clinical and radiographic findings and synovial fluid analysis were evaluated throughout the study. Macroscopic, histologic, histochemical, and biochemical findings were evaluated after necropsy. Comparisons of interest included OA and non-OA joints of saline treated horses and OA joints of PGH treated horses and OA joints of saline treated horses. Results were statistically analyzed with significance set at P < .05. RESULTS: OA caused increases in clinical assessment scores, synovial fluid variables, radiographic, macroscopic, and histologic cartilage scores, synovial fluid and cartilage chondroitin sulfate 846-epitope and glycosaminoglycan concentration. Total radiographic scores, total macroscopic joint pathology and macroscopic cartilage pathology scores were significantly reduced in horses treated with PGH compared with saline treated horses. Synovial fluid total protein concentration and white blood cell count were higher in OA joints of PGH treated horses compared with saline treated horses. There were no other significant differences between treatment groups. CONCLUSIONS: Improvements in macroscopic variables were not supported by other outcomes. Further evidence is needed before PGH can be recommended as a therapeutic option for osteoarthritis in horses.

Problems and pointers in osteochondrosis: Twenty years on.

Twenty years ago a supplement of Equine Veterinary Journal was devoted to equine osteochondrosis (OC) and recognised the importance of this developmental disease to the equine industry. In the accompanying editorial several controversial issues were identified and a number of areas for further research were highlighted. Today, equine OC is still a major clinical problem, but the on-going research has resulted in much improved knowledge and understanding of this highly complicated disease. There is still conflicting evidence on the prevalence of OC due to the dynamic character of the condition, widely varying definitions in the literature, and the range of joints affected. Nevertheless there is now convincing evidence that early vascular damage, leading to chondronecrosis, is the major mechanism of onset. The aetiological factors that determine whether a horse will develop clinical signs of OC remain obscure and the complex nature of OC and its multi-factorial character has been clearly demonstrated by genetic studies. These have shown a multitude of loci on a variety of chromosomes linked to osteochondrotic phenotypes, depending on the type of manifestation of OC, the joint involved and the breed. The controversy surrounding the possible key role of copper in the pathogenesis of OC in the early 1990s has evolved into a more limited contribution to repair thus making it just one of the many environmental factors that may have an effect on the occurrence of OC, but not a decisive one. The semantic debate concerning the most appropriate nomenclature seems to have crystallised into a consensus on terminology at three levels: OC or osteochondritis dissecans (OCD) for the disturbance in the process of endochondral ossification, juvenile ostechondral conditions (JOCC) for all joint and growth plate related disorders, and developmental orthopaedic diseases (DOD) for the full range of skeletal conditions in young horses. Future progress in improved management of OC can be expected from more research on cellular and molecular processes and the influences that determine the process of endochondral ossification, the process of articular cartilage maturation, and from epidemiological studies quantifying the long-term effects of OC on health and performance.

Effect of age and prostheses location on rima glottidis area in equine cadaveric larynges.

OBJECTIVE: To determine the effect of horse age and laryngeal prosthesis location on rima glottidis area in cadaveric larynges. STUDY DESIGN: Experimental study. ANIMALS: Cadaveric equine larynges (n = 40). METHODS: Specimens were grouped by age: group 1, ≤5 years (n = 18); group 2, >5 to ≤10 years (n = 12); group 3, >10 years (n = 10). A cranial prosthesis was placed through the dorsal cricoid spine at 70% of the distance of the total cricoid length measured from the caudal rim. A dorsal prosthesis was placed through the caudal rim of the cricoid on the dorsal midline. A lateral prosthesis was placed 1.5 cm lateral to the dorsal prosthesis. All prostheses passed through the muscular process. Rima glottidis area was determined after progressively tightening each suture in 5 N increments from 0 N to 35 N using a tensiometer. RESULTS: There was no significant effect of age on the area of the rima glottidis at any load for any of the three prosthesis locations. CONCLUSIONS: Age did not affect the area of the rima glottidis when prostheses were loaded between 5 N and 35 N.

Optimal tension, position, and number of prostheses required for maximum rima glottidis area after laryngoplasty.

OBJECTIVES: To evaluate the effect of 3 laryngeal prostheses alone or in combination on rima glottidis area in horses. STUDY DESIGN: Experimental randomized design. SAMPLE POPULATION: Cadaveric equine larynges (n = 22). METHODS: Three prostheses were preplaced in each of 14 larynges. Rima glottidis area was measured after loading each suture in 5 Newton (N) increments from 0 N to 35 N. In 8 larynges, the 3 prostheses were tied alone or in combination at a fixed load of 15 N and rima glottidis area measured. RESULTS: Rima glottidis cross-sectional area increased as the load on each prosthesis increased with maximum area reached at 20 N for each prosthesis. At a fixed load of 15 N, tying 2 and 3 prostheses in combination resulted in a larger rima glottidis cross-sectional area than achieved with each prosthesis alone. CONCLUSIONS: A combination of 2 or 3 prostheses tied at a fixed load of 15 N optimized rima glottidis cross-sectional area irrespective of the anatomic location of the prosthesis.

Altered gene expression in early osteochondrosis lesions.

Osteochondrosis is a condition involving defective endochondral ossification and retention of cartilage in subchondral bone. The pathophysiology of this condition is poorly characterized, but it has been proposed that the fundamental defect is failure of chondrocyte hypertrophy. The aim of the current study was to characterize phenotypic changes in chondrocytes associated with the initiation of osteochondrosis. Early lesions were induced in an equine model of osteochondrosis by feeding foals a high energy diet for 8 or 15 weeks. Lesions in articular-epiphyseal growth cartilage were examined histologically and by quantitative PCR analysis of expression of a number of genes representative of pathways that regulate chondrocyte behavior during endochondral ossification. There were more cells present in clusters in the lesions compared to normal articular cartilage. Expression of matrix metalloproteinase-13, type I collagen, type X collagen, and Runx2 mRNA was significantly greater in the lesions compared to normal cartilage from the same joint. Expression of vascular endothelial growth factor, type II collagen, connective tissue growth factor, aggrecan, Sox9, and fibroblast growth factor receptor 3 mRNA was not significantly different in lesions than in control cartilage. These observations suggest that osteochondrosis does not result from failure of chondrocytes to undergo hypertrophy.

Ultrasonographic evaluation of the supraspinous ligament in a series of ridden and unridden horses and horses with unrelated back pathology.

BACKGROUND: Injury to the supraspinous ligament (SSL) is reported to cause back pain in the horse. The diagnosis is based on clinical examination and confirmed by ultrasonographic examination. The ultrasonographic appearance of the supraspinous ligament has been well described, but there are few studies that correlate ultrasonographic findings with clinical pain and/or pathology. This preliminary study aims to test the hypothesis that unridden horses (n = 13) have a significantly reduced frequency of occurrence of ultrasonographic changes of the SSL consistent with a diagnosis of desmitis when compared to ridden horses (n = 13) and those with clinical signs of back pain (n = 13). RESULTS: The supraspinous ligament of all horses was imaged between T(thoracic)6-T18 and ultrasonographic appearance. There was an average of 2.08 abnormal images per horse from the whole group. The average number of abnormalities in unridden horses was 4.92, in ridden horses 2.92 and in horses with clinical back pain 4.69. No lesions were found between T6 and T10 and 68% of lesions were found between T14 and T17. No significant difference (p < 0.05) was found between the three groups in the number or location of abnormal images. CONCLUSION: The main conclusion was that every horse in this study (n = 39) had at least one site of SSL desmitis (range 2 to 11). It was clear that ultrasonographically diagnosed SSL desmitis cannot be considered as prima facie evidence of clinically significant disease and further evidence is required for a definitive diagnosis.

Effects of 25% propylene glycol hydrogel (Solugel) on second intention wound healing in horses.

OBJECTIVE: To evaluate the effect of a commercially available 25% propylene glycol hydrogel preparation (Solugel; Johnson and Johnson Medical, North Ryde, Australia) on healing of full-thickness skin wounds on the distal aspect of the limb in horses. STUDY DESIGN: Experimental. ANIMALS: Eight Standardbred horses. METHODS: Standardized (2.5 x 2.5 cm) full-thickness skin wounds were created over the mid-dorsomedial aspect of both metacarpi in 8 horses. One wound in each horse was dressed with saline solution (0.9% NaCl) soaked gauze, and one was treated with Solugel under dry regular gauze; wounds were then bandaged with gauze-coated cotton wool and elastic adhesive bandages. Wounds were videorecorded and rebandaged twice weekly until healed. Wound healing variables were measured from the videorecordings using a computer software package and analyzed as a randomized complete block design with repeated measures. Where necessary variables were made positive for analysis; significance was set at P <.05. RESULTS: The area of the wound at the first bandage change did not vary between treated and untreated wounds. Treatment had no effect on the total rate of healing, rate of healing during the retraction phase of healing, rate of healing after the retraction phase was complete, or the amount the wounds retracted. CONCLUSIONS: Using this model of wound healing, Solugel had no effect on second intention healing of distal limb wounds in horses. CLINICAL RELEVANCE: Solugel does not appear to have any beneficial effect on healing of small full-thickness skin wounds to the distal limb of horses.

The effect of equine recombinant growth hormone on second intention wound healing in horses.

OBJECTIVE: To evaluate the effect of intramuscular administration of recombinant equine growth hormone on healing of full thickness skin wounds on equine limbs. STUDY DESIGN: Experimental. ANIMALS: Nine Standardbred horses. METHODS: In study 1, standardized full thickness skin wounds (2.5 x 2.5 cm) were made over the dorsomedial aspect of the mid-cannon bone of 1 forelimb and 1 hindlimb in 9 horses. Wounds were bandaged without treatment (control subjects) and videorecorded twice weekly until healed. Then, in study 2, similar wounds were created on the opposite limbs; 6 horses were administered intramuscular recombinant equine growth hormone (10 microg/kg daily for 7 days, then 20 microg/kg daily for 49 days), and 3 horses (control subjects) were administered equivalent volumes of sterile water. Wounds were videorecorded twice weekly until healed. Wound healing variables were measured from the videorecordings using a computer software package and analyzed as a randomized complete block design factorial analysis of variance; significance was set at P <.05. RESULTS: No differences in the measured variables were detected between wounds in study 1 and the control wounds in study 2. In recombinant equine growth hormone-treated horses, wounds retracted more during treatment and contracted faster after treatment stopped when compared with wounds from untreated horses. No other treatment effects were detected. CONCLUSIONS: Recombinant equine growth hormone seemingly increases wound retraction. After treatment ceases, wound contraction increases. CLINICAL RELEVANCE: Intramuscular administration of recombinant equine growth hormone (10 microg/kg daily for 7 days, then 20 microg/kg daily for 49 days) does not appear to have any beneficial clinical effect on healing of equine limb wounds.