Distribution of TNF receptors and TNF receptor-associated intracellular signaling factors on equine tendinocytes in vitro

Although tumor necrosis factor (TNF) alpha is an important key factor in degeneration of equine superficial digital flexor tendon (SDFT), the dynamism of TNF receptors and associated factors on tendinocytes has not been elucidated. To reveal signaling events mediated by TNF-receptors (TNF-Rs) in tendinocytes, we focused on four signaling factors, TNF-R1, TNF-R2, TNF-R-associated factor 2 (TRAF2) and nuclear factor-kappa B (NF-kappaB), and investigated the distribution and production of these factors. Cultured tendinocytes were obtained from SDFTs of thoroughbred horses. The tendinocytes were treated with 10 ng/ml equine TNFalpha medium for 6 hours and then the four factors on tendinocytes were visualized by using an immunohistochemical method, and the amounts of the four factors were determined by Western blot analysis. Although TNF-R1 and TNF-R2 co-localized on the same tendinocyte, in untreated control cells (normal condition), immunoreactivity against TNF -R1 was very weak but TNF-R2 showed a strong reaction. However, TNF-R1 showed the same high level of reaction as TNF-R2 in TNFalpha-treated cells (inflamed condition). Intense TRAF2 and NF-kappaB were detected at inflamed condition, however both factors were also detected at normal condition. The distinct distributions of the four factors under different conditions (normal and inflamed condition) in vitro not only reflect the dynamism of the cytokines but may also provide important clues for a means to prevent from occurrence of tendonitis and progress of tendon degeneration.     

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.     

Effects of exercise on tenocyte cellularity and tenocyte nuclear morphology in immature and mature equine digital tendons

REASON FOR PERFORMING STUDY: The injury-prone, energy-storing equine superficial digital flexor tendon (SDFT) of the mature performance horse has a limited ability to respond to exercise in contrast with the noninjury-prone, anatomically opposing common digital extensor tendon (CDET). Previous studies have indicated low levels of cellular activity in the mature SDFT, but in foal tendons the tenocytes may still have the ability to adapt positively to increased exercise. OBJECTIVES: To measure tenocyte densities and types in histological sections from the SDFT and CDET of horses from controlled long-term, short-term and foal exercise studies. METHODS: Specimens were collected from mid-metacarpal segments of the CDET and SDFT for each horse and processed for histology; central and peripheral regions of the SDFT cross-section were analysed separately (SDFTc, SDFTp). Tenocyte nuclei were counted in a total area of 1.59 mm(2) for each tendon region in each horse. Each nucleus was classified as type 1 (elongate and thin), type 2 (ovoid and plump) or type 3 (chondrocyte-like); type 1 cells are proposed to be less synthetically active than type 2 cells. RESULTS: No significant differences were noted between exercise and control groups in any of the studies, with the exception of an exercise-related reduction in the proportion of type 1 tenocytes for all tendons combined in the long-term study. There were tendon- and site-specific differences in tenocyte densities and proportions of type 1 and 2 cells in all 3 studies. CONCLUSIONS AND POTENTIAL RELEVANCE: There was no indication that exercise increased tenocyte density or proportions of the (theoretically) more active type 2 cells in immature horses (short-term and foal studies), perhaps because the training regimens did not achieve certain threshold strain levels. In the foal study these findings can still be interpreted positively as evidence that the training regimen did not induce subclinical damage.     

A review of tendon injury: Why is the equine superficial digital flexor tendon most at risk?

Tendon injury is one of the most common causes of wastage in the performance horse; the majority of tendon injuries occur to the superficial digital flexor tendon (SDFT) whereas few occur to the common digital extensor tendon. This review outlines the epidemiology and aetiology of equine tendon injury, reviews the different functions of the tendons in the equine forelimb and suggests possible reasons for the high rate of failure of the SDFT. An understanding of the mechanisms leading to matrix degeneration and subsequent tendon gross failure is the key to developing appropriate treatment and preventative measures.     

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   

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.     

DETERMINATION OF T1 RELAXATION TIME OF NORMAL EQUINE TENDONS USING MAGIC ANGLE MAGNETIC RESONANCE IMAGING

Seven isolated equine front limbs were used to establish the normal T1 relaxation time of equine superficial digital flexor tendon (SDFT), deep digital flexor tendon (DDFT), and suspensory ligament (SL) using magic angle magnetic resonance (MR) imaging. MR imaging of the metacarpi was performed with the limbs positioned at 55° (the magic angle) relative to the main magnetic field. Transverse spin‐echo proton density and inversion recovery images were acquired. T1 relaxation time was calculated based on ratios of signal intensity determined from the different pulse sequences. T1 relaxation times for SDFT, DDFT, and SL were 288 (±17), 244 (±14), and 349 (±16) ms, respectively. The difference in T1 values between SDFT, DDFT, and SL was statistically significant. T1 values of equine tendons can be determined with magic angle imaging on a clinical MR system using <10 min total scan time. The knowledge of the normal range of T1 values may be useful to identify horses with chronic tendinopathy, where based on the human literature, an increased T1 value may be expected.     

Extracellular matrix composition of the equine superficial digital flexor tendon: relationship with age and anatomical site

The objectives of the present study were to test the hypotheses that: (1) the composition of the extracellular matrix of the equine superficial digital flexor tendon (SDFT) shows great functional similarities with articular cartilage, i.e. that significant differences exist in biochemical composition of differently loaded areas (which in the case of tendons may be more apparent as tendon shows more obvious differences than cartilage); and (2) that, as in articular cartilage, no substantial alterations in biochemical composition take place during ageing once adulthood has been attained. Tendon samples were taken from 60 adult slaughter horses from a central area at cross-section in the mid-metacarpal region and at the height of the proximal sesamoid bones (sesamoid region) of the SDFT. Contents of collagenous and non-collagenous components were determined. None of the parameters were correlated with age in either region, except for a significant increase in pentosidine cross-links with age in the sesamoid region. Between the two anatomical regions, there were significant differences in all variables, except for hydroxylysylpyridinoline cross-links. It was concluded that in the equine SDFT, similar to articular cartilage, most molecular parameters are not influenced by age in mature horses, indicating a low remodelling rate. Tendon composition is clearly different between regions, apparently reflecting different specific modes of biomechanical loading at the points sampled.     

Characterization and clinical application of mesenchymal stem cells from equine umbilical cord blood

Tendinitis of the superficial digital flexor tendon (SDFT) is a significant cause of lameness in horses; however, recent studies have shown that stem cells could be useful in veterinary regenerative medicine. Therefore, we isolated and characterized equine umbilical cord blood mesenchymal stem cells (eUCB-MSCs) from equine umbilical cord blood obtained from thoroughbred mares during the foaling period. Horses that had tendinitis of the SDFT were treated with eUCB-MSCs to confirm the therapeutic effect. After eUCB-MSCs transplantation, the core lesion in the SDFT was found to decrease. These results suggest that transplantation using eUCB-MSCs could be another source of cell treatment.     

In vitro mechanical properties of equine tendons in relation to cross-sectional area and collagen content

The mechanical properties of the deep digital flexor tendon (DDFT), the superficial digital flexor tendon (SDFT) and the suspensory ligament (SL) of the hindlimb of the horse were studied in vitro. The tendons were observed at several morphologically distinct sites. The loaded tendon is homogeneously strained, in spite of large variations in cross-sectional area. Consequently the modulus of elasticity was inversely proportional to the corresponding cross-sectional area and ranged from 738 MPa (megaPascal, N mm-2) to 1398 MPa within the DDFT, from 1000 MPa to 1282 MPa within the SDFT and from 576 MPa to 669 MPa within the SL. The collagen content was inversely proportional to the cross-sectional area and proportional to the modulus of elasticity. This stresses the influence of tendon composition on the mechanical properties, and also demonstrates the difficulty in judging the strength of a particular tendon or site within a tendon from its cross-sectional area. The respective tendons ruptured at strains of 10.0 per cent (DDFT), 12.3 per cent (SDFT) and 11.0 per cent (SL). The influence of strain rate on the modulus of elasticity is small, and these tendons may therefore be considered as non-linear elastic structures. The average hysteresis is about 5 per cent.     

Injuries of the calcaneal insertions of the superficial digital flexor tendon in 19 horses

Reasons for performing study: Injuries of the calcaneal insertions of the superficial digital flexor tendon and their relationship to displacement of the tendon from the calcaneus have not previously been reported. Objectives: To describe findings made on clinical cases with disruption of the calcaneal insertions of the superficial digital flexor tendon (SDFT) including observations on their role in horses with unstable subluxation of the tendon. To describe novel surgical techniques and the results of treatment. Hypotheses: Disruption of the calcaneal insertions of the SDFT is associated with lameness and distension of the calcaneal bursa. Unstable displacement of the SDFT from the calcaneus is a more complex injury than incomplete disruption of one of its calcaneal insertions. Methods: The case records and diagnostic images of horses with lesions involving the calcaneal insertions of the SDFT, which were confirmed by endoscopic evaluation of calcaneal bursa between 2005 and 2009, were reviewed. Results: Nineteen horses were identified including 7 that had unstable displacement of the tendon from the calcaneus. Following endoscopic surgery, 9 of 12 horses with stable tendons and 6 of 7 horses with unstable displaced SDFTs returned to work. Conclusion: Tearing of the calcaneal insertions of the SDFT is associated with lameness and distension of the calcaneal bursa; endoscopic removal of the torn tissue carries a good prognosis. Horses with unstable displacement of the tendon have also disruption of the tendon fibrocartilage cap. Removal of this results in stable subluxation and can return horses to athletic activity. Both lesions can be detected by preoperative ultrasonography. Potential relevance: Tearing of the calcaneal insertions of the SDFT should be included in the differential diagnoses of lame horses with distended calcaneal bursae. Tearing of the tendon fibrocartilage cap in horses with unstable displacement of the SDFT is a plausible explanation of the clinical features of the injury and explains previously unreliable results of reconstructive surgeries. Subtotal resection is a technically demanding technique but appears to offer an improved prognosis.     

Musculoskeletal responses of 2-year-old Thoroughbred horses to early training. 4. Morphometric, microscopic and biomechanical properties of the digital tendons of the forelimb

AIM: To determine the weight, volume, density and cross-sectional area (CSA) of the digital flexor tendons, common digital extensor tendon (CDET) and suspensory ligament (SL) of the forelimb of young Thoroughbred horses in early training, and to assess the response to a training programme of known parameters of superficial digital flexor tendon (SDFT) tissue at mid-metacarpal level. METHODS: The tendons of seven 2-year-old Thoroughbred horses in training were inspected, transected into segments of known length, and compared with those from seven untrained horses matched for age, sex and breed. The weight, volume, density and CSA of each segment, and the crimp angle, histological features, and biomechanical ultimate stress and stiffness of tendon samples from the mid-metacarpal region of the SDFT were determined. RESULTS: There was no macroscopic evidence of swelling or discolouration in any of the tendon segments or cut surfaces. The volumes of SDFT and CDET segments of horses in the trained group were significantly greater than those in the untrained group (p=0.036 and p=0.039, respectively). A greater increase in volume than weight resulted in a lower density in the SDFT but not CDET in trained compared with untrained horses (p=0.038). CSA of these two tendons was significantly greater in the trained group (p=0.002 and 0.036, respectively), the percentage increase being greater in the CDET than the SDFT. The number of tenocytes at four sites in the mid-metacarpal SDFT region was less in trained than untrained horses (p=0.025). There was no histological evidence of inflammation, and no difference in crimp angle between groups. There was no significant between-group difference in stiffness or ultimate stress of tendon strips. CONCLUSIONS: Volume and CSA of the SDFT and CDET were larger in trained than untrained horses. The SDFT was less dense in the trained group. Because no evidence of tendonitis was detected and training appeared to have no significant effect on crimp angle or biomechanical properties of tendon strips, the size and density changes were presumed to be adaptive and induced by the training. CLINICAL RELEVANCE: Although evident in this in vitro study, the detection of adaptive from initial pathological increase in size of the SDFT is likely to be difficult in vivo.     

Musculoskeletal responses of 2-year-old Thoroughbred horses to early training. 4. Morphometric,microscopic and biomechanical properties of the digital tendons of the forelimb

AIM: To determine the weight, volume, density and cross-sectional area (CSA) of the digital flexor tendons, common digital extensor tendon (CDET) and suspensory ligament (SL) of the forelimb of young Thoroughbred horses in early training, and to assess the response to a training programme of known parameters of superficial digital flexor tendon (SDFT) tissue at mid-metacarpal level.

METHODS: The tendons of seven 2-year-old Thoroughbred horses in training were inspected, transected into segments of known length, and compared with those from seven untrained horses matched for age, sex and breed. The weight, volume, density and CSA of each segment, and the crimp angle, histological features, and biomechanical ultimate stress and stiffness of tendon samples from the mid-metacarpal region of the SDFT were determined.

RESULTS: There was no macroscopic evidence of swelling or discolouration in any of the tendon segments or cut surfaces. The volumes of SDFT and CDET segments of horses in the trained group were significantly greater than those in the untrained group (p=0.036 and p=0.039, respectively). A greater increase in volume than weight resulted in a lower density in the SDFT but not CDET in trained compared with untrained horses (p=0.038). CSA of these two tendons was significantly greater in the trained group (p=0.002 and 0.036, respectively), the percentage increase being greater in the CDET than the SDFT. The number of tenocytes at four sites in the mid-metacarpal SDFT region was less in trained than untrained horses (p=0.025). There was no histological evidence of inflammation, and no difference in crimp angle between groups. There was no significant between-group difference in stiffness or ultimate stress of tendon strips.

CONCLUSIONS: Volume and CSA of the SDFT and CDET were larger in trained than untrained horses. The SDFT was less dense in the trained group. Because no evidence of tendonitis was detected and training appeared to have no significant effect on crimp angle or biomechanical properties of tendon strips, the size and density changes were presumed to be adaptive and induced by the training.

CLINICAL RELEVANCE: Although evident in this in vitro study, the detection of adaptive from initial pathological increase in size of the SDFT is likely to be difficult in vivo.     

Functional adaptation through changes in regional biochemical characteristics during maturation of equine superficial digital flexor tendons

OBJECTIVE: To quantify and compare biochemical characteristics of the extracellular matrix (ECM) of specimens harvested from tensional and compressive regions of the superficial digital flexor tendon (SDFT) of horses in age classes that include neonates to mature horses. SAMPLE POPULATION: Tendon specimens were collected on postmortem examination from 40 juvenile horses (0, 5, 12, and 36 months old) without macroscopically visible signs of tendonitis. PROCEDURE: Central core specimens of the SDFT were obtained with a 4-mm-diameter biopsy punch from 2 loaded sites, the central part of the mid-metacarpal region and the central part of the mid-sesamoid region. Biochemical characteristics of the collagenous ECM content (ie, collagen, hydroxylysylpyridinoline crosslink, and pentosidine crosslink concentrations and percentage of degraded collagen) and noncollagenous ECM content (percentage of water and glycosaminoglycans, DNA, and hyaluronic acid concentrations) were measured. RESULTS: The biochemical composition of equine SDFT was not homogeneous at birth with respect to DNA, glycosaminoglycans, and pentosidine concentrations. For most biochemical variables, the amounts present at birth were dissimilar to those found in mature horses. Fast and substantial changes in all components of the matrix occurred in the period of growth and development after birth. CONCLUSIONS AND CLINICAL RELEVANCE: Unlike cartilage, tendon tissue is not biochemically blank (ie, homogeneous) at birth. However, a process of functional adaptation occurs during maturation that changes the composition of equine SDFT from birth to maturity. Understanding of the maturation process of the juvenile equine SDFT may be useful in developing exercise programs that minimize tendon injuries later in life that result from overuse.     

Magic angle magnetic resonance imaging of diode laser induced and naturally occurring lesions in equine tendons

Magic angle magnetic resonance (MR) imaging consists of imaging tendons at 55° to the magnetic field. In people, magic angle MR imaging is valuable for detection of chronic tendon lesions and allows calculation of tendon T1 values. Increased T1 values occur in people with chronic tendinopathy. The T1 values of normal equine tendons have been reported but there are no available data for abnormal equine tendons. Twelve limbs were studied. Two limbs had diode laser tendon lesions induced postmortem, four limbs had diode laser tendon lesions induced in vivo and six limbs had naturally occurring tendon lesions. The limbs were imaged at 1.5 T using both conventional MR imaging and magic angle MR imaging. The post-mortem laser induced lesions were identified only with magic angle MR imaging. The in vivo induced lesions and naturally occurring lesions were identified with both techniques but had a different appearance with the two imaging techniques. Magic angle imaging was helpful at identifying lesions that were hypointense on conventional imaging. Increased T1 values were observed in all abnormal tendons and in several tendons with a subjectively normal MR appearance. The increased T1 value may reflect diffuse changes in the biochemical composition of tendons. Magic angle imaging has potential as a useful noninvasive tool to assess the changes of the extracellular tendon matrix using T1 values.     

Effects of platelet-derived growth factor-BB on the metabolic function and morphologic features of equine tendon in explant culture

OBJECTIVE: To evaluate the effects of recombinant human platelet-derived growth factor-BB (rhPDGF-BB) on the metabolic function and morphologic features of equine superficial digital flexor tendon (SDFT) in explant culture. Animals-6 euthanized horses (2 to 5 years old). METHODS: Forelimb SDFT explants were cultured for 6 days as untreated control specimens or treated with rhPDGF-BB (1, 10, 50, or 100 ng/mL of medium). Treatment effects on explant gene expression were evaluated via real-time PCR analysis of collagen type I, collagen type III, PDGF-A, and PDGF-B mRNA. Explants were assayed for total collagen, glycosaminoglycan, and DNA content; histologic changes were assessed via H and E staining and immunohistochemical localization of collagen types I and III. RESULTS: No morphologic or proliferative changes were detected in tendon explant sections. After high-dose rhPDGF-BB treatment, gene expression of collagen types I and III was increased and decreased, respectively. Expression of PDGF-A and PDGF-B mRNA was significantly increased at 24 hours, but later decreased to have few or negative autoinductive effects. Although PDGF gene expression waned after 48 hours of culture, collagen type I gene expression was significantly increased at 48 hours and reached peak value on day 6. Glycosaminoglycan and DNA content of explants were unchanged with rhPDGF-BB treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that rhPDGF-BB use may be of benefit in the repair of equine tendon, particularly through induction of collagen type I mRNA. Positive autoinductive effects of PDGF-BB in equine SDFT explants were detected early following culture medium supplementation, but these diminished with time.     

Are the material properties and matrix composition of equine flexor and extensor tendons determined by their functions?

REASONS FOR PERFORMING STUDY: Injury to the superficial digital flexor tendon (SDFT) is common in competition horses. The SDFT contributes to locomotory efficiency by storing energy; such tendons have low safety margins. Tendons which merely position the limb, including the opposing common digital extensor tendon (CDET), are rarely injured. The current failure of strategies to prevent or effectively treat injury to the SDFT indicates the importance of understanding how it differs from tendons which are not injury-prone. HYPOTHESIS: That the structural and material properties and matrix composition of the SDFT and CDET differ, reflecting their specific functional requirements in vivo. METHODS: Forelimb tendons were harvested from 26 mature horses and loaded to failure prior to matrix composition analysis of specimens. RESULTS: The SDFT had a significantly higher cross-sectional area, structural stiffness, failure load and failure strain and a lower elastic modulus than the CDET (P < 0.0001). CONCLUSIONS: The SDFT has conflicting requirements for strength and elasticity; although as a whole it is a stiffer structure than the CDET, differences in the matrix molecular composition including water and total sulphated glycosaminoglycan contents allow it to remain more elastic as a material. POTENTIAL RELEVANCE: Further information on how the two tendons attain these different properties may be of use in the development of prevention and treatment strategies for SDFT rupture.     

A collagenase gel/physical defect model for controlled induction of superficial digital flexor tendonitis

Reasons for performing study: A consistent and clinically relevant model for the induction of core lesions confined to the mid‐metacarpal superficial digital flexor tendon (SDFT) has not been previously reported. Injection of bacterial collagenase is commonly used but often results in large, irregular and inconsistent lesions that disrupt the superficial tendon layers and epitenon. Objective: To develop and evaluate a new injection technique for collagenase induction of SDFT injury. Methods: Collagenase gel was injected into a physical columnar defect created by longitudinally placing a curved 16 gauge 8.89 cm needle in the mid‐metacarpal SDFT in a randomly selected forelimb of 10 horses. A placebo treatment injection was performed 1 week later. Serial ultrasound examinations were performed. Horses were subjected to euthanasia at 2 (n = 2), 4 (n = 2), 8 (n = 4) and 16 (n = 2) weeks post treatment injection. Post mortem magnetic resonance imaging and histological analysis were performed. Gene expression (18S, SCX, TNC, TNMD, COL1A1, COL3A1, COMP, DCN, MMP1, MMP3 and MMP13), total DNA, glycosaminoglycan and collagen content were determined for experimental tendons (n = 10) and unaffected tendons (n = 9). Results: Mid‐metacarpal SDFT core lesion induction was successful in all tendons with consistent lesion cross‐sectional area and minimal epitenon disruption. Histology confirmed loss of normal tendon architecture after tendonitis induction and subsequent healing of the tendon core lesion. Compared with gene expression in unaffected tendons, several tested genes were significantly upregulated (COL1A1, COL3A1, TNMD, SCX, TNC, MMP13), while others showed significant downregulation (COMP, DCN, and MMP3). Conclusion: Compared with the previously used direct injection of collagenase, this injection technique was easily performed and induced more consistent lesions that were mid‐metacarpal and did not disrupt the epitenon. Potential relevance: This model will allow for objective assessment of therapies for tendon regeneration in the mid‐metacarpal SDFT prior to clinical trials and routine clinical application.     

Effects of recombinant equine growth hormone on in vitro biomechanical properties of the superficial digital flexor tendon of Standardbred yearlings in training

OBJECTIVE: To determine whether recombinant equine growth hormone (rEGH) would alter the in vitro biomechanical properties of the forelimb superficial digital flexor tendon (SDFT) in exercising young Standardbred horses. STUDY DESIGN: Randomized complete block design. ANIMALS: Twelve Standardbred yearlings. METHODS: Horses were trained for 12 weeks on a high-speed treadmill (10% positive incline). rEGH was administered intramuscularly (IM) daily (10 microg/kg during week 4; 20 microg/kg for weeks 5-9) to 6 horses (treated group), whereas 6 horses (control group) were administered an equivalent daily volume of sterile water IM. At 12 weeks, horses were euthanatized and left forelimb SDFTs were collected and stored (-70 degrees C). A section from the mid-region of the SDFT was held in cryoclamps with a 4 cm interspace distance and distracted at 10 mm/s until failure. The variables evaluated were maximal load at yield and failure, ultimate and yield tensile stress and strain, tendon stiffness, and mode of failure. Data were analyzed using unpaired, two-tailed, Student's t-test. Statistical significance was set at P < or =.05. RESULTS: Yield and ultimate tensile stress were significantly lower in the rEGH-treated horses compared with controls. There was a trend toward increased maximal displacement, increased ultimate tensile strain, and decreased tendon stiffness in rEGH-treated horses compared with controls. Tensile stress and cross-sectional area, and tensile stress and stiffness were significantly correlated at yield and failure points. CONCLUSIONS: rEGH, administered at the manufacturer's recommended dose rates to maturing Standardbred horses in training, does not significantly augment the in vitro biomechanical properties of the forelimb SDFT. CLINICAL RELEVANCE: Administration of rEGH to young horses in training is unlikely to enhance the physiologic adaptation of the SDFT to exercise stress.