The effect of recombinant equine growth hormone on the biomechanical properties of healing superficial digital flexor tendons in horses

OBJECTIVE: To evaluate the effect of recombinant equine growth hormone (rEGH) on the in vitro biomechanical properties of healing superficial digital flexor tendon (SDFT) in horses. STUDY DESIGN: Completely randomized design. SAMPLE POPULATION: Twelve Standardbred horses, 3 to 7 years of age, with ultrasonographically normal forelimb SDFT. METHODS: One week after induction of collagenase (2,000 U) induced superficial flexor tendonitis, horses were randomly divided into groups of 6. One group was administered intramuscular rEGH (10 microg/kg/day for 1 week, then 20 microg/kg/day for 5 weeks), whereas the other group (control subjects) were administered an equivalent volume of saline (0.9% NaCl) solution. At the end of this 6-week treatment, horses were killed and one forelimb SDFT from each horse was harvested for biomechanical testing under uniaxial tension. Results were analyzed using an unpaired Student's t test; significance was set at P 相似文献   

ULTRASONOGRAPHIC EVALUATION OF IATROGENIC INJURIES OF THE EQUINE ACCESSORY (CARPAL CHECK) LIGAMENT AND SUPERFICIAL DIGITAL FLEXOR TENDON

Iatrogenic lesions to the accessory ligament (carpal check or inferior check) and to the superficial digital flexor tendon of five ponies were monitored utilizing diagnostic ultrasound for a period of 6–24 weeks post-injury. The opposite limb was used as a non-surgical control. Sonographic differences from control limbs were recorded and correlated with the gross and microscopic post-mortem lesions. Early post-surgical sonographic findings included thickening with discrete hypoechoic areas within the injured tissues which corresponded to regions of hemorrhage, edema, and tissue disruption at necropsy. The linear hyperechoic pattern seen in normal tendons was disorganized in the injured portion of the tendons or ligaments. Disruption of the margins of the incised tissues was observed. The initial hypoechoic areas disappeared or decreased in size and became more echoic as healing progressed. The increasing echoic pattern in these areas corresponded to regions of fibrous tissues invasion and collagen deposition. As post-injury time increased, the size of the tendon or ligament decreased, the margins became more distinct, and the internal structure again became more organized (linear). It was concluded that diagnostic ultrasound can be useful in evaluating injury and healing of the accessory ligament and the superficial digital flexor tendon of horses.     

Healing characteristics of deep digital flexor tenorrhaphy within the digital sheath of horses

OBJECTIVE: To describe the healing characteristics of deep digital flexor tenorrhaphy within the digital sheath. STUDY DESIGN: Experimental study. ANIMALS: Five mature horses. METHODS: Right thoracic limb, deep digital flexor tenorrhaphy was performed within the digital sheath. Limbs were cast in partial flexion using a short limb cast for 6 weeks. Next, extended heel shoes were used for limb support for 14 weeks. Healing was evaluated by sequential ultrasonographic examinations, and limb use was evaluated by force plate analysis. At 26 weeks, mechanical strength and morphologic characteristics of the repair site were evaluated. RESULTS: Gap (mean, 0.93 cm.) formation was evident in unloaded limbs at 3 weeks. This increased markedly by 6 weeks and was 5 cm at 26 weeks. Demarcation between the deep and superficial flexor tendons decreased as the transected ends adhered to the dorsal surface of the superficial flexor tendon. The intrathecal space was reduced by fibrous tissue. Mean maximum load to failure of the repair tissue was 4,616 +/- 3,556 N, with a mean stress of 12.99 +/- 2.78 MPa. The repair consistently failed at the adhesion between the transected tendon and the superficial flexor tendon. CONCLUSIONS: Intrathecal tenorrhaphy with external coaptation (in partial limb flexion) for 6 weeks resulted in gap healing, fibrous adhesion between the deep and superficial flexor tendons, fibrous tissue reduction of the intrathecal space, and a pasture-sound horse at 26 weeks. CLINICAL RELEVANCE: Without improved methods for immobilizing the deep digital flexor tendon, intrathecal tenorrhaphy is unlikely to result in first intention tendon healing.     

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.     

EVALUATION OF THE EQUINE DIGITAL FLEXOR TENDON SHEATH USING DIAGNOSTIC ULTRASOUND AND CONTRAST RADIOGRAPHY

This study was designed to evaluate the normal anatomy of the digital flexor tendon sheath using contrast radiography and diagnostic ultrasound. Iodinated contrast medium was injected into eight cadaver limbs and the limbs immediately frozen. Lateromedial and dorsopalmar/plantar radiographs were made. These limps were then cut transversely and proximal to distal radiographs of each slab were made. This cross sectional contrast methodology allowed the visualization of the relative size and shape of the superficial and deep digital flexor tendons as well as the potential space taken by effusions of the digital flexor tendon sheath.
The second part of the study used twelve live animals with normal digital flexor tendon sheaths. Ultrasonographic measurement of the structures of the digital flexor tendon sheath at each level were compiled. This documented the ability of diagnostic ultrasound to image: 1) the superficial and deep digital flexor tendons, 2) the proximal and distal ring of the manica flexoria , 3) the straight and oblique sesamoidean ligaments, and 4) the mesotendinous attachments to the superficial and deep flexor tendons. Iodinated contrast medium was then injected into the digital flexor tendon sheath and the ultrasonography repeated. These images were compared with those obtained from contrast radiography and prosections of twenty normal limbs. The iodinated contrast medium enhanced sonographic imaging of the structures of the digital tendon sheath, particularly the abaxial borders of the superficial digital flexor tendon branches and the mesotendinous attachments to the superficial and deep digital flexor tendons.     

Recombinant equine growth hormone does not affect the in vitro biomechanical properties of equine superficial digital flexor tendon

OBJECTIVE: To evaluate the effect of recombinant equine growth hormone (rEGH) on the in vitro biomechanical properties of normal adult equine superficial digital flexor tendon (SDFT). STUDY DESIGN: Completely randomized design. SAMPLE POPULATION: Nine Standardbred horses, 6 to 9 years of age with ultrasonographically normal forelimb SDFT. METHODS: Six horses were administered intramuscular (IM) rEGH at 10 microg/kg/day for 1 week, and then 20 microg/kg/day for another 5 weeks; 3 horses (control subjects) were administered an equivalent daily volume of sterile water IM. Horses were killed at the end of the 6-week treatment period, and both forelimb SDFT were harvested and stored at -70 degrees C. In vitro biomechanical testing was performed under uniaxial tension. Results were analyzed using a general linear model of analysis of variance; significance was set at P <.05. RESULTS: There were no differences in cross-sectional area, maximal load at failure, yield load, ultimate and yield tensile strain, ultimate and yield tensile stress, or stiffness between tendons from control and treated horses. CONCLUSIONS: Administration of rEGH to adult Standardbred horses for 6 weeks had no detectable effect on the in vitro biomechanical properties of normal SDFT. CLINICAL RELEVANCE: Administration of rEGH does not modulate the in vitro biomechanical properties of SDFT from adult Standardbred horses.     

Cloning and expression of type III collagen in normal and injured tendons of horses

OBJECTIVE: To clone the 5' end of type III collagen and describe its pattern of mRNA and protein expression in normal and healing tendons in horses. ANIMALS: 14 healthy adult horses. PROCEDURE: The tensile region of collagenase-injured superficial digital flexor tendons was harvested at intervals from 1 to 24 weeks after injury. Total RNA was reverse-transcribed into cDNA for cloning and sequencing of type III collagen. Equine-specific nucleic acid probes were developed and used for northern blot analysis and in situ hybridization. Type III collagen protein and cyanogen bromide-cleaved collagen peptides were assessedby gel electrophresis. RESULTS: Type III collagen mRNA expression and protein content increased immediately after injury and remained increased. Type III collagen was localized to the endotenon in normal tendon and in injured tendon at 1 week. At 8 and 24 weeks, expression became more widely distributed throughout the tendon parenchyma. Injured tendon contained 6 times more type I than type III collagen mRNA. Quantities of type III collagen protein were maximal in the first 4 weeks after injury (approx 33%) and then began to decrease. CONCLUSIONS AND CLINICAL RELEVANCE: Type III collagen expression is increased initially in endotenon and subsequently in parenchyma of healing tendon; however, type III remains the minor collagen throughout the healing process. The role of type III collagen in tendon healing is not fully elucidated.     

Antemortem evaluation for magnetic resonance imaging of the equine flexor tendon

In this study antemortem evaluation of equine flexor tendons--the superficial digital flexor tendon and the deep digital flexor tendon--using magnetic resonance (MR) images was performed. Postmortem flexor tendons were used to prepare the slice positions, coil and body positions for MR imaging. It was possible by this method to take antemortem MR images of equine limbs that distinguished features as well as postmortem images described in previous studies. The total time of antemortem scanning was about 40 min. This study is the first to report antemortem MR images in horses.     

Model formulation and determination of in vitro parameters of a noninvasive method to calculate flexor tendon forces in the equine forelimb

OBJECTIVE: To describe a method to calculate flexor tendon forces on the basis of inverse dynamic analysis and an in vitro model of the equine forelimb and to quantify parameters for the model. SAMPLE POPULATION: 38 forelimbs of 23 horses that each had an estimated body mass of > or = 500 kg. PROCEDURE: Longitudinal limb sections were used to determine the lines of action of the tendons. Additionally, limb and tendon loading experiments were performed to determine mechanical properties of the flexor tendons. RESULTS: The study quantified the parameters for a pulley model to describe the lines of action. Furthermore, relationships between force and strain of the flexor tendons and between fetlock joint angle and suspensory ligament strain were determined, and the ultimate strength of the tendons was measured. CONCLUSION AND CLINICAL RELEVANCE: The model enables noninvasive determination of forces in the suspensory ligament, superficial digital flexor tendon, and distal part of the deep digital flexor (DDF) tendon. In addition, it provides a noninvasive measure of loading of the accessory ligament of the DDF tendon for within-subject comparisons. However, before application, the method should be validated. The model could become an important tool for use in research of the cause, prevention, and treatment of tendon injuries in horses.     

Healing of surgically created defects in the equine superficial digital flexor tendon: effects of pulsing electromagnetic field therapy on collagen-type transformation and tissue morphologic reorganization

The effect of pulsing electromagnetic field (PEMF) therapy on the healing of surgically created defects in equine superficial digital flexor tendons was evaluated. Defects were created in both front superficial digital flexor tendons of 20 horses. The defect in 1 limb was exposed to a PEMF for 2 hours daily. The other limb served as a control. Histologic and immunofluorescent evaluations were done in horse killed at postsurgical weeks 2, 4, 8, 12, and 24. Therapy with the PEMF significantly (P less than 0.05) delayed the maturation of the tissue formed within the defect at postsurgical weeks 8 and 12, as determined by histologic examination. The collagen-type transformation was also delayed by the PEMF therapy, but to a degree that was not significant.     

Cloning and expression of equine insulin-like growth factor binding proteins in normal equine tendon

OBJECTIVES: To define a portion of the nucleotide sequences of each of the 6 insulin-like growth factor (IGF) binding proteins (IGFBPs) in horses and describe patterns of messenger RNA (mRNA) and protein expression for IGFBPs in normal equine tendons. ANIMALS: 7 horses. PROCEDURE: Total RNA was extracted from the tensile region of normal superficial digital flexor tendons and reverse transcribed into complimentary DNA (cDNA). The cDNA was amplified via PCR, and products representing portions of each IGFBP were cloned and sequenced. Nucleotide sequences were used to deduce the amino acid sequences, and both nucleotide and predicted amino acid sequences were compared with those published for bovine, human, mouse, and ovine IGFBPs. Gene expression was quantitated by real-time PCR assay, and protein expression was evaluated by western ligand blot (WLB). RESULTS: Clones ranged in size from 262 to 522 bp and had high degrees of sequence homology with other mammalian species. Sequence homology was highest between bovine and equine IGFBPs (86% to 95%) and amongst the IGFBP-5 sequences from the various species (92% to 95%). Message for IGFBP-2 to -6, but not IGFBP-1, was expressed in normal tendon. Protein expression for IGFBP-2, -3, and -4 was detected byWLB in normal tendon and markedly increased in damaged tendons. CONCLUSIONS AND CLINICAL RELEVANCE: Results provide basic information and tools needed for further characterization of the role of the IGF system in tendon healing and may lead to the ability to potentiate the response of healing tendon to exogenous IGF-I via concurrent manipulation of IGFBPs.     

Ex vivo strength comparison of bioabsorbable tendon plates and bioabsorbable suture in a 3-loop pulley pattern for repair of transected flexor tendons from horse cadavers

OBJECTIVE: To test the failure strength and energy of 2 bioabsorbable implants applied to transected deep digital flexor tendons (DDFT) from adult horses. STUDY DESIGN: Ex vivo biomechanical experiment. SAMPLE POPULATION: Twelve pairs of deep digital flexor tendons harvested from the forelimbs of fresh equine cadavers. METHODS: Poly-L-lactic acid tendon plates were custom manufactured for application to the cylindrical surface of an adult equine deep digital flexor tendon. Twelve pairs of DDFTs were transected 2 cm distal to the insertion of the distal check ligament of the deep digital flexor tendon. One tendon of each pair was randomly selected for repair with a biodegradable plate or a 3-loop pulley method. Size 2 polydioxanone suture was used in both repairs. Repairs were tested in tension to failure, with peak force (PF) and total energy (TE) at repair failure recorded in Newtons (N) and Joules (J), respectively. A paired t-test was used for statistical evaluation with a significant level set at P< or = .05. RESULTS: Mean+/-SD PF for failure of plated tendons (1507.08+/-184.34 N) was significantly greater than for sutured tendons (460.86+/-60.93 N). TE was also significantly greater for failure of plated tendons versus sutured tendons. CONCLUSIONS: Plate fixation of transected cadaver DDFTs appear to have superior immediate failure strength than 3-loop pulley repairs. CLINICAL RELEVANCE: Whereas in vivo testing is required, a bioabsorbable tendon plate may provide initial increased strength to support tendon healing and decrease external coaptation requirements.     

Radiographic anatomy of soft tissue attachments in the equine metacarpophalangeal and proximal phalangeal region.

The sites of bony attachment of the tendons, ligaments, and fibrous portion of the joint capsules of the equine metacarpophalangeal (fetlock) joint region were determined by gross dissection. These sites were transposed to standard radiographic views of the fetlock joint to yield illustrations that can be used as an aid in the diagnosis of soft tissue pathology from radiographs. Evidence of direct attachment of the common digital extensor tendon to the proximal phalanx was not found. Branches of the superficial digital flexor tendon were found to insert only on the middle phalanx. The recently described sites of insertion of the branches of the superficial digital flexor tendon to the proximal phalanx were found to be sites for attachment of the deep axial palmar ligaments of the proximal interphalangeal joint.     

Age-related changes to the molecular and cellular components of equine flexor tendons.

Specific tendons show a high incidence of partial central core rupture which is preceded by degeneration. In the performance horse, the superficial digital flexor tendon (SDFT) is most often affected. We have described previously the molecular changes that are associated with degeneration in the central core region of the equine SDFT. The pathophysiological mechanism leading to change in synthetic activity of central zone cells in degenerated tendons is not known. In this study, we test the hypothesis that ageing results in matrix composition changes within the central zone of the SDFT. Extracellular matrix composition and cellularity were analysed in equine SDFTs collected from Thoroughbred horses and compared with a flexor tendon which rarely shows degenerative change and subsequent injury (deep digital flexor tendon, DDFT). Data were examined for age-related changes to central and peripheral zone tissue of the SDFT and DDFT. Ageing in both tendons (SDFT and DDFT) resulted in a significant increase in collagen-linked fluorescence and a decrease in cellularity in the DDFT but not the SDFT. The central zone tissue from the SDFT had a significantly higher proportion of type III collagen than the peripheral zone of the tendon. The highest level of type III collagen was found in the central zone tissue of the SDFT from the older group of horses and this may represent the early stages of a degenerative change. Collagen content did not differ between the 2 flexor tendons; however, there were differences in collagen type and organisation. The SDFT had a higher type III collagen content, higher levels of the mature trifunctional collagen crosslink hydroxylysylpyridinoline, lower total chondroitin sulphate equivalent glycosaminoglycan content, smaller diameter collagen fibrils and a higher cellularity than the DDFT. In conclusion, differences in macromolecular composition exist between the flexor tendons and ageing contributes to a tendon specific change in composition.     

Spontaneous injury of the accessory ligament of the superficial digital flexor tendon (proximal check ligament): A new ultrasonographic diagnosis

This study points out the spontaneous injury occurrence of the accessory (proximal check) ligament of the equine superficial digital flexor tendon (AL-SDFT). This injury was identified and documented ultrasonographically in 45 horses. This paper also presents the normal ultrasonographic appearance of the AL-SDFT. Ultrasonography is an adequate technique for AL-SDFT evaluation. Indications include carpal canal syndrome and proximal superficial digital flexor tendonitis.     

Evaluation of computed tomographic anatomy of the equine metacarpophalangeal joint

OBJECTIVE: To determine the detailed computed tomography (CT) anatomy of the metacarpophalangeal (MCP) joint in healthy horses. SAMPLE POPULATION: 10 cadaveric forelimbs from 10 adult horses without orthopedic disease. PROCEDURES: CT of the MCP joint was performed on 4 forelimbs. In 1 of the limbs, CT was also performed after intra-articular injection of 30 mL of contrast medium (40 mg of iodine/mL). Transverse slices 1-mm thick were obtained, and sagittal and dorsal planes were reformatted with a slice thickness of 2 mm. The CT images were matched with corresponding anatomic slices from 6 additional forelimbs. RESULTS: The third metacarpal bone, proximal sesamoid bones, and proximal phalanx could be clearly visualized. Common digital extensor tendon; accessory digital extensor tendon; lateral digital extensor tendon; superficial digital flexor tendon (including manica flexoria); deep digital flexor tendon; branches of the suspensory ligament (including its attachment); extensor branches of the suspensory ligament; collateral ligaments; straight, oblique, and cruciate distal sesamoidean ligaments; intersesamoidean ligament; annular ligament; and joint capsule could be seen. Collateral sesamoidean ligaments and short distal sesamoidean ligaments could be localized but not at all times clearly identified, whereas the metacarpointersesamoidean ligament could not be identified. The cartilage of the MCP joint could be assessed on the postcontrast sequence. CONCLUSIONS AND CLINICAL RELEVANCE: CT of the equine MCP joint can be of great value when results of radiography and ultrasonography are inconclusive. Images obtained in this study may serve as reference for CT of the equine MCP joint.     

Nucleotide structure of equine platelet-derived growth factor-A and -B and expression in horses with induced acute tendinitis

OBJECTIVE: To characterize the nucleotide sequence of equine platelet-derived growth factor (PDGF)-A and -B and analyze temporal expression of these genes in equine tendon after induced tendinitis injury. Animals-18 mature horses. PROCEDURES: Genes for equine PDGF-A and -B were reverse transcribed and sequenced from synovial tissue mRNA obtained from a 3-year-old horse. Collagenase-induced lesions were created in the tensile region of the superficial digital flexor tendon in 14 horses; 3 horses served as uninjured control animals. Tendons were harvested and total RNA was isolated from experimental horses 1, 2, 4, 8, and 24 weeks after collagenase injection. Temporal gene expression for PDGF-A and -B was determined by use of quantitative PCR analysis. RESULTS: Equine PDGF-A shared 83.8% sequence and 87.5% peptide homology with human PDGF-A, with a discrepancy of 70 bp from the human sequence. Equine PDGF-B was similar in length to the human gene, sharing 90.3% and 91.7% nucleotide and peptide identity, respectively. Expression of PDGF-A mRNA in collagenase-induced tendinitis lesions was unchanged, compared with expression for normal control tendon, and remained steady throughout the 24-week study. Expression of PDGF-B mRNA decreased over time, and the expression at 24 weeks was significantly reduced, compared with expression in normal and acutely injured tendon. CONCLUSIONS AND CLINICAL RELEVANCE: Injured tendon mounts a minimal constitutive PDGF-A or -B mRNA response. Serial exogenous treatment with either PDGF isoform within the first 2 to 4 weeks after tendon injury may bolster the meager PDGF paracrine-autocrine intrinsic response to injury.     

ULTRASONOGRAPHY OF THE EQUINE TARSUS

Ultrasonography was used to delineate the sonographic anatomy of the equine tarsus with emphasis on the flexor tendons and plantar ligament at the plantar surface of the tarsus, the medial collateral ligaments and the dorsomedial joint capsule of the tibiotarsal joint. The cross-sectional gross anatomy of these tendons and ligaments was evaluated in 6 cadaver limbs. Using a 5.5 MHz mechanical sector scanner, the examined tendons and ligaments were easily identified as hyperechoic structures in 10 limbs of normal live horses. The normal ultrasonographic appearance of the dorsomedial tibiotarsal joint capsule was studied in 8 cadaver limbs. The inner surface of the tibiotarsal joint capsule was covered with villi; short and pointed in the mid region, resembling a small nodular mass proximally and a straggly beard distally. In 4 lame horses diagnostic ultrasound aided the diagnosis of plantar swellings, medial collateral joint ligamentous injury and hypertrophic synovitis.     

Desmotomy of the accessory ligament of the superficial digital flexor tendon in the horse with use of a tenoscopic approach to the carpal sheath

Objective— To describe a tenoscopic approach to the carpal sheath for desmotomy of the accessory ligament of the superficial digital flexor tendon. Study Design— The surgical procedure was developed with use of normal forelimbs from equine cadavers and experimental horses. Animals or Sample Population— Twelve equine cadaveric forelimbs, 4 forelimbs from 2 horses anesthetized for terminal surgical laboratories, and 10 forelimbs from five experimental horses were used. Methods— The limbs were positioned lateral side up with the carpus slightly flexed. After distention of the carpal sheath, a portal was made approximately 2 cm proximal to the distal radial physis for arthroscope insertion. An instrument portal was made approximately 0.2 cm proximal to the distal radial physis. After flexion of the limb to 90°, the accessory ligament of the superficial digital flexor tendon was palpated and desmotomy was performed. Cadaveric limbs were dissected to confirm complete desmotomy. Experimental horses were monitored for short- (perioperative) and long- (4 weeks) term postoperative complications. Results— A tenoscopic approach to the carpal sheath provided adequate surgical access to the accessory ligament of the superficial digital flexor tendon for desmotomy. Most of the accessory ligament of the superficial digital flexor tendon could be easily seen within the sheath, except for the proximal 2 cm that could be readily palpated and subsequently transected. Important technical considerations were location of the arthroscope portal, adequate sheath distention, limb flexion to 90°, and desmotomy location. It was beneficial, but apparently not essential, to avoid the proximal perforating vessel. Postoperatively, some horses had swelling but were not lame and had normal range of motion of the carpus. Conclusions— Desmotomy of the accessory ligament of the superficial digital flexor tendon could be performed by using a lateral tenoscopic approach to the carpal sheath. Clinical Relevance— Desmotomy of the accessory ligament of the superficial digital flexor tendon by using a tenoscopic approach to the carpal sheath is an alternative technique to the medial incisional approach.