OBJECTIVE To investigate the effect of lipopolysaccharide (LPS) on type VII collagen– cleaving matrix metalloproteinases (MMPs) in the lamellar tissue of extracorporeally perfused equine limbs. SAMPLE 10 right forelimbs and 3 left forelimbs collected from 10 adult horses after slaughter at a licensed abattoir. PROCEDURES Extracorporeal perfusion of the isolated equine limbs was performed for 10 hours under physiologic conditions (control-perfused limbs; n = 5) and with the addition of 80 ng of LPS/L of perfusate (LPS-perfused limbs; 5). Lamellar tissue specimens were then collected from the dorsal aspect of the hooves. Additionally, corresponding control specimens were collected from the 3 nonperfused left forelimbs. Immunohistochemical analysis was performed on paraffin-embedded tissue blocks with antibodies against total (latent and active) MMP-1, MMP-2, MMP-8, and MMP-9 as well as antibody against active MMP-9. Intensity of immunohistochemical staining was scored, and stain distribution in the lamellar tissue was noted. RESULTS Staining intensity of total and active MMP-9 was significantly increased in LPS-perfused versus control-perfused limbs. No such difference was identified for MMP-1, MMP-2, and MMP-8. CONCLUSIONS AND CLINICAL RELEVANCE Of the 4 MMPs that are capable of degrading type VII collagen, MMP-9 was the only one for which production increased in the lamellar tissue of isolated equine limbs perfused with versus without a clinically relevant concentration of LPS. These results suggested that MMP-9 may be involved in initiation of pathological changes in lamellar tissue in endotoxin-induced laminitis, whereas MMP-1, MMP-2, and MMP-8 may be less relevant.

The digital cushion (DC) plays a role in absorbing and dampening forces applied to the foot and therefore supports internal structures such as navicular bone; yet, its architecture is not well-known. The goal of this study was to characterize the microanatomical structure of the DC in horses with clinically sound hooves. Both forefeet from the cadavers of 12 adult Quarter horses were cut and sectioned and samples of the following 4 regions of the DC were obtained: axial proximal (AxProx), axial distal (AxDis), abaxial lateral (AbxLat), and abaxial medial (AbxMed). The samples were processed and stained with hematoxylin and eosin, Masson's trichrome, and Weigert's elastic stains. On each slide, 2 central 3- × 3-mm areas were microscopically assessed and all measurements were done within the 9-mm2 area. The number of detected collagen bundles, nerve fascicles, vessels, and the diameter of wall thickness and lumen of blood vessels were measured. Elastic fiber profiles were categorized based on relative density of elastic fibers detected in the field. The percentage of samples in which chondrocytes and adipose tissues were either present or absent was calculated. Significant structural differences were identified among the 4 regions of the DC. The AxDis region contained more collagen bundles (P < 0.0001) and less elastic fiber profiles than the AxProx region (P < 0.0001). The AxDis also contained more collagen bundles than the AbxMed and AbxLat (P < 0.0001) regions. Our findings provide insight into the structure of the DC of mature Quarter horses. The structural differences in the various regions of the DC are presumably related to the different functional properties of those regions; yet more research is warranted.

OBJECTIVE To determine the effects of racetrack surface and shoe characteristics on formation of wear grooves in the horseshoes of racehorses. SAMPLES 1,121 horseshoes from 242 Thoroughbred racehorses collected during routine horseshoeing procedures at 4 racetracks with dirt or synthetic surfaces. PROCEDURES Data for 1,014 horseshoes from 233 racehorses were analyzed. Horseshoes were photographed, and length and width of grooves formed at the heels of the solar surface of horseshoes were measured on the photographs. Effects of racetrack, racetrack surface, and shoe characteristics (eg, shoe size, clips, and nails) on length and width of grooves were assessed by use of a mixed-model anova. RESULTS Length and width of wear grooves differed significantly on the basis of racetrack, nail placement, and limb side (left vs right). Differences in groove dimensions between types of racetrack surface (dirt vs synthetic) were less apparent than differences among racetracks. CONCLUSIONS AND CLINICAL RELEVANCE Measurements of the length and width of wear grooves in the horseshoes of racehorses may be useful for understanding some aspects of hoof interactions with various racetrack surfaces. Interpretation of differences in wear grooves for various racetrack surfaces will likely require quantitation of the mechanical behavior of the surfaces.

Objective—To determine intralimb orientation changes with an inertial measurement unit (IMU) in hooves of horses at a walk and trot after induction of weight-bearing single forelimb lameness and to determine whether hoof orientations are similar to baseline values following perineural anesthesia. Animals—6 clinically normal horses. Procedures—3-D hoof orientations were determined with an IMU mounted on the right forelimb hoof during baseline conditions, during 3 grades of lameness (induced by application of pressure to the sole), and after perineural anesthesia. Linear acceleration profiles were used to segment the stride into hoof breakover, stance, initial swing, terminal swing, and total swing phases. Intralimb data comparisons were made for each stride segment. A repeated-measures mixed-model ANOVA was used for data analysis. Results—Lameness resulted in significant changes in hoof orientation in all planes of rotation. A significant increase in external rotation and abduction and a significant decrease in sagittal plane rotation of the hoof were detected at hoof breakover during lameness conditions. For sagittal plane orientation data, the SDs determined following perineural anesthesia were higher than the SDs for baseline and lameness conditions. Conclusions and Clinical Relevance—Results of this study indicated the IMU could be used to detect 3-D hoof orientation changes following induction of mild lameness at a walk and trot. An increase in data variability for a sagittal orientation may be useful for assessment of local anesthesia for hooves. The IMU should be further evaluated for use in clinical evaluation of forelimb lameness in horses.

Objective: To validate an equine inertial measurement unit (IMU) system rigidly attached to a hoof against a 3-D optical kinematics system in horses during walking and trotting. Animals: 5 clinically normal horses. Procedures: 5 swing phases of the hooves of the right forelimb and hind limb were collected via both 3-D optical and IMU systems from 5 horses during walking and trotting. Linear and angular positions, velocities, and accelerations were compared between the 2 systems. Results: Of the 55 variables compared between the 2 systems, 25 had high correlations (r > 0.8) and 18 had moderate correlations (r > 0.5). Root mean squared errors were lowest in the sagittal plane and orientation (1.1 to 4.4 cm over a range of 1.5 to 1.9 m in the cranial-caudal direction and 2.5° to 3.5° over a range of 88° to 110° rotating around the medial-lateral axis). There were more differences between the 2 systems during small changes in motion, such as in the medial-lateral and proximal-distal directions and in the angular measures around the cranial-caudal and proximal-distal axes. Conclusions and Clinical Relevance: The equine IMU system may be appropriate for rigid attachment to a hoof of a horse and use in examination of linear and angular motion in the sagittal plane of the hoof during the swing phase while walking and trotting. Although promising in many respects, the IMU system cannot currently be considered clinically useful for lameness evaluation because of limitations in accuracy, attachment method, and lack of stance phase evaluation.

Objective: To determine the expression and distribution of a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), its substrates aggrecan and versican, and their binding partner hyaluronan in laminae of healthy horses. Sample: Laminae from the forelimb hooves of 8 healthy horses. Procedures: Real-time quantitative PCR assay was used for gene expression analysis. Hyaluronidase, chondroitinase, and keratanase digestion of lamina extracts combined with SDS-PAGE and western blotting were used for protein and proteoglycan analysis. Immunofluorescent and immunohistochemical staining of tissue sections were used for protein and hyaluronan localization. Results: Genes encoding ADAMTS-4, aggrecan, versican, and hyaluronan synthase II were expressed in laminae. The ADAMTS-4 was predominantly evident as a 51-kDa protein bearing a catalytic site neoepitope indicative of active enzyme and in situ activity, which was confirmed by the presence of aggrecan and versican fragments bearing ADAMTS-4 cleavage neoepitopes in laminar protein extracts. Aggrecan, versican, and hyaluronan were localized to basal epithelial cells within the secondary epidermal laminae. The ADAMTS-4 localized to these cells but was also present in some cells in the dermal laminae. Conclusions and Clinical Relevance: Within digital laminae, versican exclusively and aggrecan primarily localized within basal epithelial cells and both were constitutively cleaved by ADAMTS-4, which therefore contributed to their turnover. On the basis of known properties of these proteoglycans, it is possible that they can protect the basal epithelial cells of horses from biomechanical and concussive stress.

Objective—To quantify changes in hoof wall strain distribution associated with exercise and time in Standardbreds. Animals—18 young adult Standardbreds. Procedures—9 horses were exercised 4 d/wk for 30 to 45 minutes at a medium trot for 4 months; 9 nonexercised horses served as the control group. Rosette strain gauges were used to measure the principal surface strains at the toe, lateral quarter of the hoof wall (LQ), and medial quarter of the hoof wall (MQ) of the right forefoot at the beginning and end of the experiment. Midstance maximal (msepsilon1) and minimal (msepsilon2) principal and peak minimal principal (pkepsilon2) surface strains were measured; SDs of each of those variables were also calculated. Results were compared through ANOVA of time and exercise effects between and within the groups. Results—Both the exercised and nonexercised groups had changes in strain distribution in their hooves over time. The msepsilon1 did not change significantly with exercise; however, it changed significantly in both groups at both hoof quarters over time. At the beginning of the study, mean msepsilon2 and pkepsilon2 values were significantly higher in the exercised group than in the control group at the MQ and LQ but not at the toe. At the end of the study, these values were significantly higher in the control group than in the exercised group at the toe but not at the MQ or LQ. Conclusions and Clinical Relevance—Detected changes in hoof wall surface strain may indicate the ability of hoof capsule material to respond to exercise. A better understanding of hoof adaptation to applied forces may allow implementation of proper trimming and shoeing techniques to promote adaptation to exercise loads in horses.