Light and electron microscopy of keratinization in the laminar epidermis of the equine hoof with reference to laminitis

The laminar epidermis (epidermis parietis) of hooves from 14 clinically normal horses, 6 months to 15 years old, was examined by light and electron microscopy and immunofluorescence to measure the contributions of this region to the formation of the hoof wall. By their progressive keratinization to form primary epidermal laminae, the secondary epidermal laminae ultimately contributed about 20% of the thickness of the hoof wall (as revealed in the white line [zona alba]). The keratinized, primary epidermal laminae were developed to a height of 4 mm during their proximodistal-course, much of this obscured because of their basal portion being embedded in the cap horn epidermis. From evaluation of structural changes accompanying keratinogenesis in the cell and determination of the contribution of the laminar epidermis to the formation of laminar horn, cap horn, connecting horn, terminal horn, and the white line, we concluded that the sterile bed concept of a nongerminative role for the secondary epidermal laminae is no longer tenable.     

Evaluation of architectural changes along the proximal to distal regions of the dorsal laminar interface in the equine hoof

OBJECTIVE: To describe architectural changes along the dorsal laminar interface of the equine foot. SAMPLE POPULATION: 6 macroscopically normal forefeet obtained from 6 equine cadavers. PROCEDURE: Histologic sections of 8 evenly spaced, proximal to distal, samples of the dorsal laminar interface were photographed, digitized, and examined for differences in architecture. Laminar depth; secondary laminar density; number and consistency of bifurcations occurring within the secondary laminae, and areas composed of primary dermal lamina, primary epidermal lamina, and secondary laminar interface were recorded. Data were examined to test for differences in architecture associated with the proximal to distal positioning of the sample. RESULTS: With exception of the areas of the primary epidermal and primary dermal lamina, all measured variables were significantly different between the proximal and distal regions of the dorsal laminar interface. Changes included increases in laminar depth and the secondary laminar density. Bifurcation of secondary laminae principally occurred proximally and had an increased depth of bifurcation distally. The secondary laminar dermal-epidermal interface had a 109% increase in area between the most proximal and distal sections. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this study indicate that the interface normally contributes a substantial volume of dermal components to the internal surface of the wall. These data also indicate that 2 distinct mechanisms (i.e., bifurcation of secondary laminae and an increase in the length of secondary laminae) contribute to changes in the architecture of the laminar interface, which allows for the hypothesis that the normal laminar interface is capable of responding to mechanical load.     

Characterisation and distribution of epidermal growth factor receptors in equine hoof wall laminar tissue: comparison of normal horses and horses affected with chronic laminitis

Epidermal growth factor (EGF) receptors were detected in plasma membrane preparations of equine hoof wall laminar tissue at concentrations comparable to that of equine liver. Scatchard analysis of the equilibrium binding data suggested the presence of two classes of EGF binding sites in most of the controls (plasma membranes from clinically normal horses); a high-affinity class and a more numerous low-affinity class. The dissociation constant of the low-affinity class of EGF-specific receptors (KD = 1 x 10(-9)M) is in reasonable agreement with other values established for the EGF receptor. The variability between individual estimates for the KD of the high-affinity receptor class precluded an accurate estimate for those sites. A possible explanation is discussed. The high-affinity binding sites were uniformly absent in plasma membranes prepared from horses affected by chronic laminitis. Autoradiographic analysis localised the EGF receptors primarily to the secondary epidermal laminae, with an apparent greater density over the proliferative basal keratinocytes. Little label was associated with the dermal or the keratinised primary epidermal laminae. Tissue from horses with chronic laminitis had EGF receptors located uniformly over the hyperplastic epidermal keratinocytes. These data suggest that an EGF-mediated response may be involved in the hyperproliferative response that is characteristic of chronic laminitis.     

Density and morphologic features of primary epidermal laminae in the feet of three-year-old racing Quarter Horses

OBJECTIVE: To quantify the density of primary epidermal laminae (PELs) around the solar circumference and evaluate the relationship between regional PEL density and hoof capsule morphology in horses. SAMPLE POPULATION: Forefeet from nine 3-year-old Quarter Horse cadavers. PROCEDURES: Data pertaining to gross features of hoof morphology and PEL variables, including number, density, and distribution patterns around the perimeter of the hoof wall and number of bar PELs, were collected. Tissues of the laminar junction were examined histologically. RESULTS: No significant differences were found between left and right forefeet with respect to gross hoof morphologic measurements. Mean +/- SD number of PELs, including those at the bars of the hoof, was 551 +/- 30. Primary epidermal laminar density in the toe was significantly higher, compared with that in the quarter and heel regions, and was higher on the lateral aspect of the foot, compared with the medial aspect. Feet were significantly wider on the lateral aspect of the foot, compared with the medial aspect, as measured across the widest point of the solar surface. Histologic examination revealed atypical laminar morphology at the toe. CONCLUSIONS AND CLINICAL RELEVANCE: Variations were detected in PEL density and morphology around the solar circumference of hooves from 3-year-old racing Quarter Horses. A better understanding of relationships between laminar density, laminar morphology, and gross morphology of the hoof capsule in different populations of horses may aid practitioners in diagnosis and treatment of disease involving the hoof wall in horses.     

Morphological spectrum of primary epidermal laminae in the forehoof of Thoroughbred horses

Reasons for performing the study: Hoof health is a major concern of horse owners as well as the equine industry. However, many questions remain concerning regional variations of laminar junction and its potential to remodel. Hypothesis: To examine regional variations in the morphology of the laminar junction and thickness of the hoof wall in Thoroughbred horses. Methods: The forefeet of 25 Thoroughbred cadavers were examined. Each hoof was divided into 20 blocks through 4 proximodistal slices (below the coronary band, each 1 cm apart) and 5 circumferential positions (toe, medial and lateral quarters and heels). In each block, 25 central primary epidermal laminae (PEL) were considered. Orientation of each lamina in relation to the hoof wall (LO), degree of bending (IA) and the spaces between the adjacent laminae (LS) were measured. Thickness of the hoof wall and number of branched PEL were also measured. Data were analysed using a split‐block design in ANOVA. Results: There were significant differences between the 2 proximal and 2 distal slices in LO and IA data, but not in LS data. Circumferentially, toe blocks were different from heel and quarters blocks. Lateral and medial heels as well as the quarters were mostly different. The hoof wall was slightly thicker laterally than medially. There were more branched PEL on the lateral side of the left hooves and on the medial side of the right hooves. Conclusions: These data add to the circumstantial evidence supporting the hypothesis of adaptive remodelling in the laminar junction. Results of this study signify the capability of PEL to remodel in response to applied stress to the regions of the hoof. Potential relevance: A deeper understanding of the gross and cellular processes of laminar remodelling may well prove to be complementary to an understanding of their failure in laminitis.     

Development of intraosseous infusion of the distal phalanx to access the foot lamellar circulation in the standing,conscious horse

Intraosseous (IO) infusion of the distal phalanx (IOIDP) as a delivery route targeting hoof lamellar tissue of standing, conscious horses was evaluated. Following sedation and regional nerve blockade in six Standardbred horses, a microdialysis (MD) probe was implanted into the hoof lamellar tissue of one forelimb. A purpose designed cannulated bone screw was introduced into the body of the distal phalanx, approximately 6 cm from the MD probe. Gentamicin solution (25 mg/mL) was infused at 20 μL/min through the bone screw for 2 h without the application of a tourniquet. MD and blood samples were collected at regular intervals and analysed for gentamicin concentrations.Gentamicin was present in lamellar tissue at much higher concentrations than peripheral serum. The mean concentration of gentamicin was 24.4, 20.5 and 4.4 μg/mL in extracellular fluid (ECF) and 0.28, 0.5 and 0.32 μg/mL in serum samples collected 60, 120 and 150 min after IOIDP was started, respectively. A clinically safe and efficacious IO drug delivery to the hoof lamellar tissue of standing, conscious horse was developed.     

Decreased expression of p63, a regulator of epidermal stem cells, in the chronic laminitic equine hoof

Reasons for performing study: Abnormal epidermal stem cell regulation may contribute to the pathogenesis of equine chronic laminitis. Objective: To analyse the involvement of p63, a regulator of epidermal stem cell proliferative potential, in chronic laminitis. Methods: Epidermal tissues from skin, coronet and lamellae of the dorsal foot were harvested from 5 horses with chronic laminitis and 5 control horses. Tissues were analysed using histopathology, immunofluorescence microscopy and quantitative immunoblotting Results: Hoof lamellae of laminitic horses had a lower frequency of p63 positive cells than control lamellae, particularly in the distal region. Quantitative immunoblotting confirmed reduced p63 expression in the laminitic distal lamellar region. The decreased p63 expression in laminitic epidermal lamellae was most apparent in the abaxial region adjacent to the hoof wall and highly associated with the formation of terminally differentiated, dysplastic and hyperkeratotic epidermis in this region, whereas lamellae from control horses maintained high p63 expression throughout the axial‐abaxial axis. Conclusions: Expression of p63 in equine skin resembles that reported in other species, including man and rodents, suggesting that p63 can serve as a marker for the proliferative potential of equine epidermal stem cells. p63 expression was significantly lower in the chronic laminitic hoof than in that of control horses, suggesting laminitic hoof epithelium has more limited proliferative potential with a shift towards differentiation. This may reflect reduced activity of epidermal stem cells in laminitic hoof. It is proposed that p63 contributes to the maintenance of hoof lamellae and that misregulation of p63 expression may lead to epidermal dysplasia during lamellar wedge formation. Potential relevance: This study suggests that loss of epidermal stem cells contributes to the pathogenesis of equine laminitis. Autologous transplantation of p63‐positive epidermal stem cells from unaffected regions may have regenerative therapeutic potential for laminitic horses.     

Quantitative morphology of the equine laminar junction in relation to capsule shape in the forehoof of Standardbreds and Thoroughbreds

REASONS FOR PERFORMING STUDY: Differences in hoof morphology have largely been underappreciated in the literature until recently, and it is these that hold the key to interpreting functional adaptation in the hoof. HYPOTHESES: Primary laminar morphology correlates with hoof capsule shape; and breeds with different hoof shapes and loadings show different patterns of correlation. METHODS: Seventeen measurements of capsule shape and 3 of primary epidermal laminae (PEL) morphology (spacing, orientation and curvature) were made on right and left front hooves from 27 Standardbred and 25 Thoroughbred horses, and tested for breed differences. Three laminar variables (spacing, orientation and curvature) were measured on each hoof for samples of 25 PEL in 5 circumferential and 4 proximodistal locations. Pairwise correlations of capsular and laminar measurements were compared within breeds. Significant correlations were mapped onto the 20 sampling sites. RESULTS: Capsule shape differed significantly between breeds in 7 measurements and in a multivariate test. Between breeds, PEL differed in orientation and spacing primarily at the medial quarters and heels, and in curvature at both quarters (P<0.05). Significant correlations between several pairs of capsule and laminar variables were found at sample locations that differed markedly between breeds. CONCLUSIONS: Laminar morphology, hoof capsule shape and correlations between them differ between Standardbreds and Thoroughbreds. These results support the concept that remodelling of PEL is, at least in part, stimulated and directed by varying stress or strain levels in the laminar junction. POTENTIAL RELEVANCE: Understanding the biological responses of hoof tissues to stress should add to the ability to prevent lameness involving the hoof and maintain its health.     

Assessment of apoptosis in epidermal lamellar cells in clinically normal horses and those with laminitis

OBJECTIVE: To determine and compare the number, type, location, and distribution of apoptotic epidermal cells in the laminae of clinically normal horses and horses with laminitis. SAMPLE POPULATION: Formalin-fixed samples of digital lamellar tissue from 47 horses (including clinically normal horses [controls; n = 7], horses with acute [4] and chronic [7] naturally acquired laminitis, and horses with black walnut extract-induced [11] or carbohydrate overload-induced [18] laminitis). PROCEDURE: Blocks of paraffin-embedded lamellar tissues were stained for DNA fragmentation with the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) technique. Differential immunohistochemical staining for caspases 3 and 14 were used to confirm apoptosis. RESULTS: The number of TUNEL-positive epidermal cells per 0.1 mm of primary laminae was significantly greater in the acute laminitis group than in the other groups. In the acute laminitis group, there were 17 and 1,025 times as many TUNEL-positive basal layer cells and keratinocytes, respectively, compared with the control group. Apoptosis of TUNEL-positive basal layer cells was confirmed by results of caspase 3 immunohistochemical staining. The TUNEL-positive keratinocytes did not stain for caspases 3 or 14. CONCLUSIONS AND CLINICAL RELEVANCE: The large number of apoptotic basal layer cells detected in the lamellar tissue of horses with acute naturally acquired laminitis suggests that apoptosis may be important in the development of acute laminitis. The role of the large number of TUNEL-positive keratinocytes detected in the interface of primary and secondary epidermal laminae of horses with acute laminitis remains to be elucidated.     

Biomechanical and histopathological changes in the support structures of bovine hooves around the time of first calving

Hooves were removed from heifers killed two weeks before calving (C-2), and four and twelve weeks post-calving (C+4 and C+12), and from age-matched maiden heifers. Segments were dissected from the anterior walls of lateral hind claws, to include horn, corium and bone, and the biomechanical properties were measured. There was reduced rigidity, both overall and at a physiologically appropriate displacement of 2 mm, and a greater displacement was required to reach initial and maximum support in the hooves of calving, compared with maiden, heifers. Histological examination revealed widened and distorted interdigitating laminae, progressing to C+12 in the hind claws, but with evidence of recovery in the front claws after C+4. Increased connective tissue strength from dorsal to ventral hoof segments was correlated with protein, proteoglycan, pro- and activated matrix metalloproteinase-2, and tissue inhibitors of metalloproteinases, and was inversely correlated with fat, water and collagen content. This implies that mechanical changes reflect alterations in the biochemistry of the connective tissue. This evidence supports the hypothesis that primary causal events associated with calving weaken the connective tissue of the hoof suspensory apparatus, leading to increased susceptibility to clinical lameness associated with sole ulcers and white line disease.     

Equine laminitis caused by distal displacement of the distal phalanx: 12 cases (1976-1985)

Clinical data from 12 cases of equine laminitis characterized by distal displacement of the distal phalanx (P3) were reviewed. Clinical features of horses that survived the syndrome were compared with the nonsurvivors to obtain prognostic indicators. Animals affected included 8 Quarter Horses, 2 Welsh ponies, 1 Thoroughbred, and 1 Arabian. Eight of the animals were females (67%), 2 were stallions, and 2 were geldings. The mean age of affected animals was 8.6 years (2 to 14 years), and the mean body weight was 442 kg. The survivors weighed less than the nonsurvivors (384 kg vs 473 kg, respectively), suggesting that body weight may be of prognostic value for horses affected with distal displacement of P3. Ten of the 12 animals (83%) were admitted because of a disorder other than laminitis, but subsequently developed laminitis during the treatment period. All affected animals had clinical evidence of endotoxemia and/or sepsis before the onset of laminitis. Cavitation or depression of the dorsal coronary band was detected in all animals and was the most reliable clinical indicator of distal displacement of P3. Five horses had fluid (blood or serum) ooze from their coronary bands and 2 of these sloughed one or more of their hooves. Necropsy findings of the 8 horses that were euthanatized included severe hemorrhagic, congested laminae and complete detachment of P3 from the hoof wall. Histologic examination of affected laminae revealed vascular thrombosis and multifocal areas of hemorrhage and necrosis. Radiography failed to reveal distal displacement of P3 in 8 animals, but the remaining 4 animals had an accentuation of the dorsal proximal hoof wall and cavitation of the coronary band visible on lateral radiographs.     

Fenestration of the abaxial hoof wall and implantation of gentamicin-impregnated collagen sponges for the treatment of septic arthritis of the distal interphalangeal joint in cattle.

A novel surgical approach to the distal interphalangeal joint of cattle, through the abaxial hoof wall, for the treatment of septic arthritis is described. In seven cattle a rectangular piece of horn (15 x 40 mm) was excised from the abaxial hoof wall and the joint was lavaged with Ringer's solution. Infected and/or necrotic tissue was removed from the joint and gentamicin-impregnated collagen sponges were introduced. The piece of horn was reattached, a rubber block was glued to the contralateral claw, and a bandage applied. Aftercare consisted of changes of bandage, systemic administration of antimicrobial drugs and box rest. At follow-up, six of the cattle were sound but one cow which had been slaughtered five months after surgery had remained slightly lame.     

INTRAARTERIAL CONTRAST-ENHANCED COMPUTED TOMOGRAPHY OF THE EQUINE DISTAL EXTREMITY

A technique for regional delivery of contrast medium to the foot of the horse to increase the diagnostic utility of computed tomography (CT) for the characterization of soft tissue structures within the hoof capsule was developed. An intraarterial catheter was placed under ultrasound guidance into the medial palmar artery at the level of the carpus to facilitate a steady-state infusion of iodinated contrast medium. Two 5 mm collimated contiguous acquisition CT scans were performed in 10 horses without lameness or radiographic evidence of distal sesamoid bone degeneration. CT examination was performed before and during regional arterial infusion of iodinated contrast medium administered at a rate of 2 ml/s. Cross-sectional area and mean pre- and post-contrast attenuation of the deep digital flexor tendon and the collateral ligaments of the distal interphalangeal joint were quantified from CT images. Soft tissue anatomy of the foot was also qualitatively characterized from pre- and postcontrast images. Catheterization was successful and without complication in all horses. The evaluated tendons and ligaments were clearly visible and had a small (8-20 Hounsfield Unit) but significant (P < 0.05, paired t-test) increase in attenuation during the steady-state infusion of contrast medium. This study should enhance the diagnostic potential of CT by providing baseline quantitative data for comparison with horses affected with soft tissue injuries in the distal extremity causing lameness that is alleviated with a palmar digital nerve block.     

Ultrastructure of the horse tongue: further observations on the lingual integumentary architecture

This investigation examined primarily epidermal specializations of the adult horse tongue by light, scanning and transmission electron microscopy. Samples were collected from seven regions of the normal tongue of various breeds of horse. The filiform papillae, present on the dorsal and lateral aspects but not the ventral aspect of the tongue, were short, slender and finger-like structures with variable-shaped terminae. The epidermal thickness and height of dermal ridges were reduced on fungiform and vallate papillae, but tissue architecture and keratinocyte ultrastructure of most of the lingual epidermis corresponded to the common mammalian epidermal paradigm. One unique finding was the highly localized clustering of epidermal cells with exceptionally high content of PAS-negative trichohyalin cytoplasmic granules at a location atop the dermal ridges and beneath the base of filiform papillae. These granular cells were immediately subjacent to clusters of clear, non-granulated epidermal cells. It is believed that this integumentary specialization may enhance the structural strength at this localized site of the tissue architecture, in relationship to the mechanical papillae.     

A gross and histopathological study of an ectopic white line development in equine laminitis

In horses with chronic laminitis, an abnormal horny structure called lamellar wedge, is generated between the hoof wall and the laminar epidermis. To be able to manage horses with chronic laminitis correctly, more information about the pathological state of this abnormal horn is required. The aim of this study was to collect and analyze objective morphological data about the abnormal horn in order to understand its morphology and development. In the study, the abnormal horn was grossly visible on the sagittal hoof section from approximately 20 days after the onset of disease. In the histological observations, the structural characteristics of this abnormal horn were similar to the white line tissue, suggesting it is an ectopic white line. Mean value of the cross-sectional area of the abnormal horn against the distal phalanx section area (A/D) was 0.29 cm(2) SD +/- 0.14 and it finally showed an eight-fold increase over the mean value of normal white line section area against the distal phalanx section area. In conclusion, a large amount of the ectopic white line is thought to be finally able to inhibit normal hoof wall growth, so that it should be resected at the optimum time when would be after one month from the onset of the disease.     

Magnetic resonance microscopy of the equine hoof wall: a study of resolution and potential

REASONS FOR PERFORMING STUDY: Obtaining magnetic resonance images of the inner hoof wall tissue at the microscopic level would enable early accurate diagnosis of laminitis and therefore more effective therapy. OBJECTIVES: To optimise magnetic resonance imaging (MRI) parameters in order to obtain the highest possible resolution of the structures beneath the equine hoof wall. METHODS: Magnetic resonance microscopy (MRM) was performed in front feet from 6 cadaver horses using T2-weighted fast spin echo (FSE-T2), and T1-weighted gradient echo (GRE-T1) sequences. RESULTS: In T2 weighted FSE images most of the stratum medium showed no signal, however the coronary, terminal and sole papillae were visible. The stratum lamellatum was clearly visible and primary epidermal lamellae could be differentiated from dermal lamellae. CONCLUSION: Most structures beneath the hoof wall were differentiated. Conventional scanners for diagnostic MRI in horses are low or high field. However this study used ultra-high field scanners currently not available for clinical use. Signal-to-noise ratio (S/N) increases as a function of field strength. An increase of spatial resolution of the image results in a decreased S/N. S/N can also be improved with better coils and the resolution of high field MRI scanners will increase as technology develops and surface array coils become more readily available. POTENTIAL RELEVANCE: Although MR images with microscopic resolution were obtained ex vivo, this study demonstrates the potential for detection of lamellar pathology as it occurs. Early recognition of the development of laminitis to instigate effective therapy at an earlier stage and may improve the outcome for laminitic horses. Clinical MR is now readily available at 3 T, while 4 T, 7 T and 9 T systems are being used for human whole body applications.     

Equine laminitis: Ultrastructural lesions detected in ponies following hyperinsulinaemia

Reasons for performing study: Anatomical changes in the hoof lamellar tissue induced by prolonged hyperinsulinaemia have not been described previously. Analysis of the induced lesions may promote understanding of hyperinsulinaemic laminitis pathogenesis and produce clinical benefit. Objectives: To use light and transmission electron microscopy (TEM) to document hoof lamellar lesions in ponies clinically lame after prolonged hyperinsulinaemia. Methods: Nine clinically normal, mature ponies were allocated randomly to either a treatment group (n = 5) or control group (n = 4). The treatment group received insulin via a modified, prolonged euglycaemic hyperinsulinaemic clamp technique (EHCT) and were subjected to euthanasia when clinical signs of Obel grade II laminitis occurred. The control group was sham treated with an equivalent volume of 0.9% saline and killed at 72 h. Lamellar tissues of the right front feet were harvested and processed for TEM. Results: Lamellae from insulin treated ponies were attenuated and elongated with many epidermal basal cells (EBC) in mitosis. Unlike carbohydrate induced laminitis in horses there was no global separation at the lamellar dermal/epidermal interface among ponies. Sporadic EBC basement membrane (BM) separation was associated with the proximity of infiltrating leucocytes. In 2 ponies, the lamellar BM was thickened. The number of hemidesmosomes/μm of BM was decreased in all insulin treated ponies. Conclusions: Prolonged hyperinsulinaemia causes unique lamellar lesions normally characteristic of acute and chronic laminitis. Lamellar proliferation may be an insulin effect through its mitogenic pathway. Aberrant lamellar mitosis may lengthen and weaken the lamellar, distal phalanx attachment apparatus and contribute to the clinical signs that developed. Potential relevance: The study shows that insulin alone, in higher than normal circulating concentrations, induces profound, changes in lamellar anatomy. Medical control of insulin resistance and hyperinsulinaemia may ameliorate lesions and produce clinical benefit.     

A preliminary study into the correlation of stiffness of the laminar junction of the equine hoof with the length density of its secondary lamellae

Reason for performing study: The relationship between mechanical behaviour and microscopic structure of the laminar junction of equine hooves under testing conditions requires elucidation. Objectives: To determine mechanical parameters and 2D length density of profiles of secondary lamellae of the laminar junction in the dermal region and to assess possible correlations. Methods: Specimens (25 samples in total) of the laminar junction were taken from front, quarter and heel parts from 3 equine hooves and exposed to a uniaxial tensile test until rupture to obtain Young's moduli of elasticity, ultimate stress and strain. Neighbouring specimens to those used for the biomechanical experiment were processed histologically to assess the length density of laminar junction basement membrane using stereological grids. Results: The estimated median (interquartile range) length density of the laminar junction basement membrane was 0.024 (0.020–0.027)/µm. Young's modulus of elasticity was 0.15 (0.11–0.35) MPa in the small deformation region, and 7.58 (6.14–8.68) MPa in the linear region was. The ultimate stress was 1.67 (1.41–2.67) MPa, and the ultimate strain was 0.50 (0.38–0.70). The Young's modulus of elasticity in the region of small deformations has a moderate correlation with the length density of the laminar junction basement membrane. Conclusions: As with most soft biological tissues, the laminar junction has a nonlinear mechanical behaviour. Within the range of small deformations, which correspond to physiological loading of the laminar junction, a higher length density of the laminar junction basement membrane is correlated with a higher resistance of the laminar junction against high stresses transmitted from the distal phalanx to the hoof wall. Potential relevance: The condition of the laminar junction apparatus may be easily quantified as the length density of profiles of secondary dermal lamellae. This quantification provides a simple tool that could be used for comparing the proneness of the various parts of the laminar junction to initial stages of laminitis.     

Accuracy of open magnetic resonance imaging for guiding injection of the equine deep digital flexor tendon within the hoof

Lesions of the distal deep digital flexor tendon (DDFT) are frequently diagnosed using MRI in horses with foot pain. Intralesional injection of biologic therapeutics shows promise in tendon healing; however, accurate injection of distal deep digital flexor tendon lesions within the hoof is difficult. The aim of this experimental study was to evaluate accuracy of a technique for injection of the deep digital flexor tendon within the hoof using MRI‐guidance, which could be performed in standing patients. We hypothesized that injection of the distal deep digital flexor tendon within the hoof could be accurately guided using open low‐field MRI to target either the lateral or medial lobe at a specific location. Ten cadaver limbs were positioned in an open, low‐field MRI unit. Each distal deep digital flexor tendon lobe was assigned to have a proximal (adjacent to the proximal aspect of the navicular bursa) or distal (adjacent to the navicular bone) injection. A titanium needle was inserted into each tendon lobe, guided by T1‐weighted transverse images acquired simultaneously during injection. Colored dye was injected as a marker and postinjection MRI and gross sections were assessed. The success of injection as evaluated on gross section was 85% (70% proximal, 100% distal). The success of injection as evaluated by MRI was 65% (60% proximal, 70% distal). There was no significant difference between the success of injecting the medial versus lateral lobe. The major limitation of this study was the use of cadaver limbs with normal tendons. The authors conclude that injection of the distal deep digital flexor tendon within the hoof is possible using MRI guidance.