Computerized ultrasonographic tissue characterization of equine superficial digital flexor tendons by means of stability quantification of echo patterns in contiguous transverse ultrasonographic images

OBJECTIVE: To describe a method of computerized ultrasonographic tissue characterization that includes structures below the size limits of resolution in equine superficial digital flexor tendons. SAMPLE POPULATION: 2 damaged and 2 structurally normal superficial digital flexor tendons. PROCEDURE: Transverse ultrasonographic images were collected along the tendon long axis. Stability of echo pattern was quantified by means of variation in gray levels of each pixel in contiguous images and expressed as correlation, entropy, and waviness ratios. RESULTS: Normal young and normal old tissues were characterized by high correlation and low entropy and waviness ratios. In necrotic tissue, collapsed intratendinous septa resulted in high correlation, moderate entropy, and high waviness ratios. In early granulation tissue, complete lack of bundle formation resulted in values of zero for correlation and waviness ratios; loose connective tissue matrix resulted in a high entropy ratio. In late granulation tissue, formation of new bundles resulted in a high correlation ratio; swollen intratendinous septa and incomplete organization of connective tissue matrix were reflected in high entropy and waviness ratios. In early fibrotic tissue, rearrangement of tendon bundles resulted in a correlation ratio within reference range and a slight increase in the waviness ratio; an increase in cellularity and lack of fibrillar arrangement led to an increase in the entropy ratio. In late fibrotic and scar tissues, inferior quality of repair with almost complete lack of organization was reflected in low to moderate correlation, low waviness, and high entropy ratios. CONCLUSIONS AND CLINICAL RELEVANCE: Stability of echo patterns accurately reflects homogeneity of tendons in horses.     

ULTRASONOGRAPHIC EVALUATION OF EQUINE TENDONS: A QUANTITATIVE IN VITRO STUDY OF THE EFFECTS OF AMPLIFIER GAIN LEVEL, TRANSDUCER-TILT, AND TRANSDUCER-DISPLACEMENT

The objective of the in vitro experiments described in this paper was to quantify the effects of some instrumental variables on the quantitative evaluation, by means of first-order gray-level statistics, of ultrasonographic images of equine tendons. The experiments were done on three isolated equine superficial digital flexor tendons that were mounted in a frame and submerged in a waterbath. Sections with either normal tendon tissue, an acute lesion, or a chronic scar, were selected. In these sections, the following experiments were done: 1) a gradual increase of total amplifier gain output subdivided in 12 equal steps; 2) a transducer tilt plus or minus 3 degrees from perpendicular, with steps of 1 degree; and 3) a transducer displacement along, and perpendicular to, the tendon long axis, with 16 steps of 0.25 mm each. Transverse ultrasonographic images were collected, and in the regions of interest (ROI) first-order gray-level statistics were calculated to quantify the effects of each experiment. Some important observations were: 1) the total amplifier gain output has a substantial influence on the ultrasonographic image; for example, in the case of an acute lesion, a low gain setting results in an almost completely black image; whereas, with higher gain settings, a marked "filling in" effect on the lesion can be observed; 2) the relative effects of the tilting of the transducer are substantial in normal tendon tissue (18%) and chronic scar (12%); whereas, in the event of an acute lesion, the effects on the mean gray level are dramatic (40%); and 3) the relative effects of displacement of the transducer are small in normal tendon tissue, but on the other hand, the mean gray-level changes 7% in chronic scar, and even 20% in an acute lesion. In general, slight variations in scanner settings and transducer handling can have considerable effects on the gray levels of the ultrasonographic image. Furthermore, there is a strong indication that this quantitative method, as far as based exclusively on the first-order gray-level statistics, may be not discriminative enough to accurately assess the integrity of the tendon. Therefore, the value of a quantitative evaluation of the first-order gray-level statistics for the assessment of the integrity of the equine tendon is questionable.     

Structure-related echoes in ultrasonographic images of equine superficial digital flexor tendons

OBJECTIVE: To develop a method to discriminate between structure-related echoes and echoes resulting from interference, as observed in transverse ultrasonographic images of equine superficial digital flexor (SDF) tendons. SAMPLE POPULATION: 2 normal (injury-free) SDF tendons obtained from a 3-year-old Thoroughbred and a 9-year-old Dutch Warmblood horse. PROCEDURE: Tendons were mounted in a custom-made device that permitted exact transverse and perpendicular sequential scanning with precise steps of 0.5 mm along the long axis of the tendon. Photographs of transverse tendon sections at the exact scanning locations were obtained. Propagation, reflection, and refraction artifacts were quantified, and an image rectification procedure was developed, allowing exact matching of each photograph with the corresponding ultrasonographic image. A correlation routine was developed that departed from this transverse ultrasonographic image (position 0); this routine added information from images collected at precise distances of 0.5 and 1 mm on both sides of the actual scan location (positions -2, -1, +1, +2). RESULTS: By use of the correlation routine, echoes that remained steady over all 5 images were enhanced and resolved, and constantly changing echoes were multiplicatively reduced and faded. This correlated image could be projected over the rectified photograph, and the resolved echoes matched perfectly with the endotendon septa surrounding fibers and fasciculi. CONCLUSIONS AND CLINICAL RELEVANCE: The correlation routine permits exclusive resolution of structure-related echoes, as echoes resulting from interference are faded. The technique described can produce images that depict only the essential structure-related information. In this way, the clinical assessment of tendon integrity is greatly facilitated.     

Efficacy of computerized discrimination between structure-related and non-structure-related echoes in ultrasonographic images for the quantitative evaluation of the structural integrity of superficial digital flexor tendons in horses.

OBJECTIVE: To evaluate effectiveness of computerized discrimination between structure-related and non-structure-related echoes in ultrasonographic images for quantitative evaluation of tendon structural integrity in horses. SAMPLE POPULATION: 4 superficial digital flexor tendons (2 damaged tendons, 2 normal tendons). PROCEDURE: Transverse ultrasonographic images that precisely matched histologic sections were obtained in fixed steps along the long axis of each tendon. Distribution, intensity, and delineation of structure-related echoes, quantitatively expressed as the correlation ratio and steadiness ratio , were compared with histologic findings in tissue that was normal or had necrosis, early granulation, late granulation, early fibrosis, or inferior repair. RESULTS: In normal tendon, the even distribution of structure-related echoes with high intensity and sharp delineation yielded high correlation ratio and steadiness ratio. In areas of necrosis, collapsed endotendon septa yielded solid but blurred structure-related echoes (high correlation ration and low steadiness ratio). In early granulation tissue, complete lack of organization caused zero values for both ratios. In late granulation tissue, reorganization and swollen endotendon septa yielded poorly delineated structure-related echoes (high correlation ratio, low steadiness ratio). In early fibrosis, rearrangement of bundles resulted in normal correlation ration and slightly low steadiness ratio. In inferior repair, the almost complete lack of structural reorganization resulted in heterogeneous poorly delineated low-intensity echoes (low correlation ratio and steadiness ratio). CONCLUSIONS AND CLINICAL RELEVANCE: The combination of correlation ratio and steadiness ratio accurately reflects histopathologic findings, making computerized correlation of ultrasonographic images an efficient tool for quantitative evaluation of tendon structural integrity.     

The effects of polyester (terylene) fibre implants on normal equine superficial digital flexor tendon

AIM: To determine the effect of polyester (terylene) fibre implants on normal equine superficial digital flexor (SDF) tendon structure. METHODS: Normal forelimb SDF tendons (n=24) of 12 horses were divided into unoperated, sham-operated, and implanted (terylene fibre implant) groups. Horses were assessed for lameness and ultrasonographic changes to SDF tendons at intervals up to 48 weeks post-operatively. After euthanasia, SDF tendons were collected for histological and ultrastructural examination. Histological sections were examined for alcian blue staining intensity, cellularity, fibril bundle alignment, fascicle separation and crimp morphology. Mass-average diameters (MADs) of collagen fibrils were calculated from electron micrographs and compared between treatment groups. RESULTS: Insertion of terylene fibre implants resulted in short-term (8 weeks) lameness in implanted limbs. Ultrasonographically, the implants could be detected in 50% of implanted tendons, but were associated with tendon swelling and the presence of hypoechoic core lesions in 7/8 implanted limbs. There were significant alterations in alcian blue staining, cellularity and crimp morphology in the central fascicles of sham-operated and implanted tendons, and alteration in fibril alignment in the central fascicles of implanted tendons. Unoperated tendons remained histologically normal. MADs of collagen fibrils did not differ between sham-operated, implanted and unoperated limbs. CONCLUSIONS: Both the sham procedure and the insertion of terylene fibre implants led to alterations in tendon structure that persisted for up to 48 weeks. Persistence of disorganised connective tissue at the proximal and distal ends of the terylene fibre implants may predispose implanted tendons to continued risk of injury. CLINICAL RELEVANCE: It is unlikely that terylene fibre implants offer any advantage over standard non-surgical treatments for mild to moderate cases of SDF tendonitis in the horse.     

EVALUATION OF SUPERFICIAL DIGITAL FLEXOR TENDINITIS IN RACING HORSES THROUGH GRAY SCALE HISTOGRAM ANALYSIS OF TENDON ULTRASONOGRAMS

Normal and diseased superficial digital flexor tendons at different stages of healing from seven Thoroughbred horses, aged 3–6 years, were evaluated by correlation of gray scale histogram analysis of tendon cross-sectional ultrasonograms with histopathologic examination. Normal tendon and tendons at different stages of healing, excluding acute tear stage, were characterized by normally distributed histogram patterns. The histogram mean value of normal tendons was higher than the corresponding values in diseased tendons at different stages of healing (p < 0.01). In addition, the histogram mean of acute lesions was lower than for early fibrosis or healed lesions (p < 0.01). The histogram mean increased as healing progressed. The results show that gray scale histogram analysis of tendon ultrasonogram can be used to distinguish between normal and abnormal superficial digital flexor tendon in racing horses.     

Magnetic resonance imaging application to live horse for diagnosis of tendinitis

Six live horses with various stages of acute to chronic superficial digital flexor (SDF) tendinitis were examined using magnetic resonance imaging (MRI). In each case, MRI findings were compared to the corresponding ultrasonographic (USD) and histologic findings, to establish the usefulness of MRI. In the acute cases, lesions characterized by hemorrhage were well defined as high signal intensity on MRI and hypoechoic regions on USD. Chronic tendon fibrosis was slightly hyperechoic and difficult to distinguish from the normal tendon tissue around the original injury by using USD. In contrast, MRI visualized the chronic lesion as a low intensity signal, which could be distinguished from the black background of the normal SDF tendon tissue. This study clearly demonstrated MRI was the better imaging modality for the objective detection of chronic scar tissue in live horses. These findings, from living horses, suggest an advantage of MRI in the clinical application to diagnose tendinitis in cases where there is chronic scar tissue that is difficult to discern on USD.     

ULTRASONOGRAPHY OF THE DORSAL AND LATERAL ASPECTS OF THE EQUINE CARPUS: TECHNIQUE AND NORMAL APPEARANCE

The normal ultrasonographic appearance of the dorsal and lateral soft tissue structures and anatomic landmarks of the equine carpus useful in clinical imaging are described.
Both limbs of 5 cadavers and 5 clinically sound adult horses were imaged using a 7.5 MHz sector transducer. At the dorsal aspect of the carpus and distal radius, the extensor carpi radialis- and the common digital extensor tendon and their tendon sheaths were easily identified. These two tendons are the most prominent structures in this region. Smaller and more difficult to identify are the tendon and tendon sheath of the extensor carpi obliquus-, the lateral digital extensor- and the ulnaris lateralis muscle. The ultrasonographic appearance and course through the carpal region of the tendons and tendon sheaths mentioned are described. Other soft tissue structures examined include the lateral collateral ligament, the carpal joint capsule and the distal articular cartilage of the radius. Ultrasonographic findings correlated well with gross anatomy in the cadavers limbs.     

ULTRASONOGRAPHIC ANATOMY OF THE NORMAL CANINE CALCANEAL TENDON

To provide a series of detailed ultrasonographic images of the canine calcaneal tendon, greyhound cadaver pelvic limbs were scanned with a high-resolution transducer, and images compared with dissected specimens. The three components of the calcaneal tendon are the tendons of insertion of the gastrocnemius, superficial digital flexor, and a conjoined tendon formed by tendons from the biceps femoris, semitendinosus, and gracilis. Each of these three tendons was visible in transverse ultrasound images, each measuring 2.4-3.2-mm thick at mid-calcaneal tendon. Improved understanding of the anatomy of the calcaneal tendon will support clinical ultrasonography of this region.     

EFFECT OF PERINEURAL ANESTHESIA ON THE ULTRASONOGRAPHIC APPEARANCE OF EQUINE PALMAR METACARPAL STRUCTURES

The purpose of this study was to describe ultrasonographic changes of the equine palmar metacarpal area attributed to the infiltration of local anesthetic solution and to determine whether these changes were noted immediately or at 24 h. The palmar metacarpal region of one forelimb in each of six horses was examined ultrasonographically with a 10-MHz linear array transducer and a 7.5-MHz curvilinear transducer. Transverse and longitudinal images were recorded at 5-cm intervals distal to the accessory carpal bone. High and low palmar and palmar metacarpal nerve blocks were performed with a 2% mepivacaine hydrochloride solution. Ultrasonographic examinations similar to the initial examination then were performed immediately, 1 h and 24 h postinjection. Cross-sectional area and mean pixel value were determined for the superficial and deep digital flexor tendons, the accessory ligament of the deep digital flexor tendon, the suspensory ligament, and the suspensory branches at each level and time period. Subjective ultrasonographic changes also were noted. No significant difference was noted in the cross-sectional area or mean pixel value of any structure at any level or time period compared to baseline. Subjective changes in the tendons and ligaments were not noted. There was mild hypoechoic swelling of the surrounding soft tissues and gas in the region of the injections. Gas could interfere with the evaluation of the origin of the suspensory ligament and the proximal portion of the accessory ligament of the deep digital flexor tendon within the first hour but was not detectable ultrasonographically at 24 h. Based on these findings, if gas interferes with an ultrasonographic examination performed temporally close to perineural anesthesia, a repeat examination at 24 h is recommended.     

Tendonitis of the branches of insertion of the superficial digital flexor tendon in horses

Objective To describe clinical findings, ultrasonographic features and outcome of injury to the branches of insertion of the superficial digital flexor (SDF) tendon in horses.
Design Retrospective study of 14 cases.
Procedure Fourteen Thoroughbred horses with tendonitis affecting the branches of insertion of the SDF tendon were examined for lameness, location and amount of swelling, and the presence of other musculoskeletal abnormalities. The flexor tendons were assessed by ultrasonographic examination, and recommendations were made for management of the cases. Outcome was assessed by re-examination of some horses, direct communication with the owner or trainer, and examination of race records.
Results The lateral SDF branch was affected in 10 horses; the medial branch in three, and both branches in one horse. Two horses had concurrent injuries to the SDF tendon in the metacarpal region of the contralateral limb. Ultrasonographic findings included swelling of the affected SDF branch, peritendinous fluid accumulation, disruption of normal fibre alignment on sagittal scan, and variable loss of echogenicity. As healing occurred, there was return of normal echogenicity, but normal fibre alignment did not return completely and apparent adhesions formed between the affected SDF branch and adjacent structures. Seven of 10 horses which returned to their previous use were able to compete without further tendon injury. Recurrence of injury occurred in one case, and another two horses developed tendonitis in the metacarpal region. One horse was retired from racing but was able to compete at dressage without recurrence of injury. Two horses were retired for breeding without returning to training, and one horse was sold and lost to follow up but did not race.
Conclusion The prognosis is fair for return to previous use following injury to the branches of insertion of the SDF tendon in athletic horses.     

ULTRASONOGRAPHY OF THE EQUINE SHOULDER: TECHNIQUE AND NORMAL APPEARANCE

This study was intended to document normal ultrasonographic appearance of the equine shoulder and anatomic landmarks useful in clinical imaging. Both forelimbs of five equine cadavers and both forelimbs of six live adult horses were used. To facilitate understanding of the images, a zoning system assigned to the biceps brachii and to the infraspinatus tendon was developed. Ultrasonography was performed with a real-time B-mode semiportable sector scanner using 7.5- and 5-MHz transducers. On one cadaver limb, magnetic resonance imaging (MRI) was performed using a system at 1.5 Tesla, T1-weighted spin-echo sequence. Ultrasonography images were compared to frozen specimens and MRI images to correlate the ultrasonographic findings to the gross anatomy of the shoulder. Ultrasonography allowed easy evaluation of the biceps brachii and the infraspinatus tendon and their bursae, the supraspinatus muscle and tendons, the superficial muscles of the shoulder, and the underlying humerus and scapula. Only the lateral and, partially, the caudal aspects of the humeral head could be visualized with ultrasound. Ultrasonographic appearance, orientation, and anatomic relationships of these structures are described. Ultrasonographic findings correlated well with MRI images and with gross anatomy in the cadavers' limbs.     

ULTRASONIC ANATOMY OF THE TENDONS AND LIGAMENTS IN THE DISTAL METACARPAL-METATARSAL REGION OF THE EQUINE LIMB

In a two-phase study, ultrasound was used to delineate the normal sonographic anatomy of soft tissues of the equine distal limb. The study was limited to the soft tissues of the palmar surface of the limb just proximal to the fetlock joint. In the first phase, cadavers were evaluated with a B-mode ultrasound machine, † Then the limbs were radiographed and dissected to compare their gross, radiographic, and ultrasonographic appearances. In the second phase, nine normal adult horses were ultrasonographically scanned. The sonographic appearance of the normal animals was compared with that of the cadavers. Front and rear limbs of all horses had similar ultrasonographic appearances. Flexor tendons and the suspensory ligament were easily identified as having linear, uniformly intense ecogenicity. The borders of the digital sheath and of the proximal palmar pouch of the fetlock joint had a less organized and less intense ecogenic appearance than the flexor tendons and suspensory ligament, and they were poorly delineated unless distended with fluid. Ultrasonography was used in three lame horses and aided the diagnoses of synovial proliferation in the fetlock joint, tendosynovitis, and bowed tendon. Diagnostic ultrasonography was effective in identifying and characterizing soft-tissue structures of the distal equine limb. It is a valuable, noninvasive diagnostic aid.     

Acellular Urinary Bladder Matrix in a Collagenase Model of Superficial Digital Flexor Tendonitis in Horses

The objective of the present study was to determine the efficacy of urinary bladder matrix (UBM) in collagenase-induced superficial digital flexor (SDF) tendonitis by using clinical, ultrasonographic, and histologic data. A total of eight healthy adult horses were used in this study. Bilateral forelimb SDF tendonitis was created in the horses by injecting collagenase. After 14 days, one randomly selected forelimb SDF tendon was blindly treated with UBM and the opposite tendon was treated with a control (saline). Clinical and ultrasonographic parameters including lameness, lesion size, ultrasonographic fiber pattern, and echogenicity were measured throughout the study. After 84 days, horses were euthanized and SDF tendon lesions from the two groups were compared statistically using an analysis of variance with significance set at P ≤ .05.Results showed that there were no significant differences between the treated and control tendons for any of the clinical, ultrasonographic, gross, or histologic variables. UBM does not appear to be an effective treatment for collagenase-induced SDF tendonitis. However, there may be differences in clinical tendonitis that might render the treatment more effective in the clinical setting.     

Preliminary study of tendon biopsy in the horse

A series of experimental tendon biopsies is described. Three biopsies were taken from the lateral digital extensor tendon (LDET) and three from the superficial flexor tendon (SFT). The LDET biopsies resulted in little discomfort whereas the SFT biopsies led to temporary lameness. The tendons were examined histologically up to 99 days after the biopsies were removed. In all of the tendons the defect filled with granulation tissue which subsequently became organised as a longitudinally orientated collagenous scar tissue. In this small series of biopsies the histological effects of the biopsy persisted longer in the SFT than in the LDET. It is concluded that tendon biopsy cannot be advocated for safe routine use in the horse.     

Ultrasound appearance of the palmar metacarpal soft tissues of the horse

Using real-time ultrasound imaging, the normal anatomy of the soft tissues of the palmar aspect of the equine forelimb was examined. Sections of frozen specimens were prepared which correlated well with the ultrasound images. The superficial and deep digital flexor tendons, tendon sheaths, inferior check ligament (accessory ligament of the deep digital flexor tendon), suspensory ligament, contours of the apices of the proximal sesamoids and the intersesamoidean ligament, were identified.     

Injury Induces a Change in the Functional Characteristics of Cells Recovered from Equine Tendon

Injury initiates a repair process characterized by influx of fibroblasts and the rapid formation of fibrous scar tissue and subsequent tissue contraction. The response to injury and behavior of the different tendon fibroblast populations, however, has been poorly characterized. We hypothesized that the fibroblasts recovered from tendon with acute injury would exhibit different cell properties relating to adhesion, migration and tensegrity. To test this hypothesis we evaluated the ability of fibroblasts recovered from normal and injured equine superficial digital flexor tendons (SDFTs). The injured tendon-derived cells showed greater contraction of the collagen gel but poorer adhesion to pepsin-digested collagen, and migration over extracellular matrix proteins compared to normal SDFT-derived fibroblasts. Thus, the cells present within the tendon after injury display different behavior related to wound healing.     

Effects of Superior Check Desmotomy on Flexor Tendon and Suspensory Ligament Strain in Equine Cadaver Limbs

OBJECTIVE: To measure the effects of transection of the accessory ligament of the superficial digital flexor (SDF) muscle (superior check desmotomy) on flexor tendon and suspensory ligament (SL) strain in vitro. STUDY DESIGN: In vitro experimental biomechanical investigation. ANIMALS USED: Ten equine cadaver forelimbs. METHODS: The effects of superior check desmotomy were determined using equine cadaver forelimbs secured in a servocontrolled hydraulic testing machine. Strain sensors were used to measure strain on the superficial and deep digital flexor tendons and SL, and a goniometer was used to measure joint angles when the limb was loaded at 890 N and 3,115 N before desmotomy, and at 3,115 N after desmotomy. RESULTS: Superior check desmotomy was associated with significantly increased strains on the SDF tendon and SL, and significant alterations in the angles of the metacarpophalangeal and carpal joints. CONCLUSIONS: The superior check ligament has an important role in maintaining joint angles and load distribution in the forelimb. Lengthening of the SDF musculotendinous unit after superior check desmotomy may be associated with increased strain on the SL. CLINICAL RELEVANCE: Transection of the accessory ligament of the SDF muscle may predispose horses to SL desmitis postoperatively.     

Use of quantitative analysis of sonographic brightness for detection of early healing of tendon injury in horses

OBJECTIVES: To determine whether quantitative analysis of sonographic brightness could be used to detect healing of an induced injury of the superficial digital flexor tendon in horses and whether rate of healing was influenced by equine recombinant growth hormone. ANIMALS: 8 clinically normal Standardbreds. PROCEDURES: A localized injury was created in the left and right superficial digital flexor tendons of each horse by injection of 2,000 units of collagenase. After injury, 4 horses received equine recombinant growth hormone, a possible promoter of tendon healing. Sonographic images (7.5 MHz) of the flexor tendons and ligaments of the metacarpal region were recorded on videotape prior to injury and weekly for 7 weeks after injury. Images were digitized, and sonographic brightness of tendons and ligaments was calculated. RESULTS: Collagenase-induced injury was sonographically similar to naturally occurring injury. After injury, sonographic brightness of the tendon decreased; after 3 weeks, brightness progressively increased, although by 7 weeks brightness had not returned to preinjury value. Equine recombinant growth hormone had no significant effect on the rate of tendon healing, as evaluated sonographically or at necropsy. CONCLUSIONS AND CLINICAL RELEVANCE: As healing developed, alterations in sonographic brightness of injured tendons coincided with real changes in tendon structure. Quantitative sonographic brightness could be used to accurately monitor healing of equine tendon and ligament injuries and investigate the efficacy of various treatment regimens.