Thomas H. Witte

Determination of peak vertical ground reaction force from duty factor in the horse (Equus caballus).

SUMMARY Measurement of peak vertical ground reaction force (GRFz) from multiple limbs simultaneously during high-speed, over-ground locomotion would enhance our understanding of the locomotor mechanics of cursorial animals. Here, we evaluate the accuracy of predicting peak GRFz from duty factor (the proportion of the stride for which the limb is in contact with the ground). Foot-mounted uniaxial accelerometers, combined with UHF FM telemetry, are shown to be practical and accurate for the field measurement of stride timing variables, including duty factor. Direct comparison with the force plate produces a mean error of 2.3 ms and 3.5 ms for the timing of foot on and foot off, respectively, across all gaits. Predictions of peak GRFz from duty factor show mean errors (with positive values indicating an overestimate) of 0.8±0.04 N kg–1 (13%; N=42; mean ± s.e.m.) at walk, –0.3±0.06 N kg–1 (3%; N=75) at trot, –2.3±0.27 N kg–1 (16%; N=18) for the non-lead limb at canter and +2.1±0.7 N kg–1 (19%; N=9) for the lead limb at canter. The substantial over- and underestimate seen at canter, in the lead and non-lead limbs, respectively, is attributed to the different functions performed by the two limbs in the asymmetrical gaits. The difference in load experienced by the lead and non-lead limbs decreased with increasing speed.

Gene therapy in musculoskeletal repair.

Abstract:  Local and regional gene therapy has improved healing in preclinical trials of articular and other muculoskeletal conditions. Combinations of cell supplementation and cells overexpressing growth factor genes have shown promising results for improving cartilage repair, enhancing delayed union of fractures, and driving organized tendon repair. Proof of concept has been developed using viral vectors, predominantly adenovirus, to deliver growth factor genes, such as BMP‐2, TGF‐β1, and IGF‐I. Integrating vectors, such as retrovirus and lentivirus, have improved the duration of gene‐induced repair, however, increased risk factors have limited broad application. Cartilage repair can be improved using chondrocyte or stem cell transplantation with cells expressing IGF‐I, BMP‐2, or FGF‐2. In cartilage injury and secondary osteoarthritis models, a combination of IL‐1 knockdown and growth factor supplementation has restored cartilage matrix and stabilized the osteoarthritic process. Ultimately, nonviral vectors may provide similar control of catabolic activity in cartilage and synovial structures, which may further improve outcome after chondrocyte or mesenchymal stem cell (MSC) implantation. MSCs derived from bone marrow, fat, or other connective tissues provide a multipotent cell source that may be privileged vectors for skeletal gene therapy. MSCs expressing BMP‐2, TGF‐β1, LMP‐1, IGF‐I, or GDF‐5 have enhanced cartilage, bone, and tendon repair. Overall, the field of orthopedic gene therapy for enhanced tissue repair has made significant preclinical advances. Combining existing cell transplant technology to deliver differentiated cells in a minimally invasive way, with genes that improve matrix formation, provides a manageable protocol for a persisting anabolic impact.

Centre of mass movement and mechanical energy fluctuation during gallop locomotion in the Thoroughbred racehorse

SUMMARY During locomotion cyclical interchange between different forms of mechanical energy enhances economy; however, 100% efficiency cannot be achieved and ultimately some mechanical work must be performed de novo. There is a metabolic cost associated with fluctuations in mechanical energy, even in the most efficient animals. In this study we investigate the exchanges between different forms of mechanical energy involved in high-speed gallop locomotion in Thoroughbred race horses during over-ground locomotion using innovative, mobile data collection techniques. We use hoof-mounted accelerometers to capture foot contact times, a GPS data logger to monitor speed and an inertial sensor mounted over the dorsal spinous processes of the fourth to sixth thoracic vertebrae (the withers) of the horse to capture trunk movement with six degrees of freedom. Trunk movement data were used to estimate the movement of the centre of mass (CoM). Linear (craniocaudal, mediolateral and dorsoventral) and rotational (roll, pitch and heading) kinematic parameters (displacement, velocity and acceleration) were calculated for seven horses at gallop speeds ranging from 7 to 17 m s-1 during their regular training sessions. These were used to estimate external mechanical energy (potential energy and linear kinetic energy of the CoM) as well as selected components of internal energy (angular kinetic energy). Elastic energy storage in the limbs was estimated from duty factor, sine wave assumptions and published leg stiffness values. External mechanical energy changes were dominated by changes in craniocaudal velocity. Potential energy change, which was in phase with craniocaudal energy during the front limb stances, was small. Elastic energy storage in the limbs was small compared to the overall amplitude of fluctuation of external mechanical energy. Galloping at high speeds does not therefore fit classical spring mass mechanics.

Cytokine and catabolic enzyme expression in synovium, synovial fluid and articular cartilage of naturally osteoarthritic equine carpi.

REASONS FOR PERFORMING STUDY Understanding the expression of catabolic and anabolic genes during osteoarthritis progression should help to identify the major mediators of the disease. OBJECTIVE To compare the cytokine and anabolic marker concentrations in synovium, synovial fluid and cartilage between normal and osteoarthritic joints. METHODS Carpi from horses age 2-11 years were used. Tissues were harvested at the time of surgery or euthanasia, and RNA was isolated for RT-PCR analysis. Tumour necrosis factor alpha (TNFα), interleukin-1beta (IL-1β), aggrecanase 1 (ADAMTS-4), aggrecanase 2 (ADAMTS-5), matrix metalloproteinase-13 (MMP-13), interleukin 17 (IL-17) and collagen type I alpha 1(Col-1) expression were determined in synovium. TNFα, IL-1β, ADAMTS-4, ADAMTS-5, MMP-13, IL-17, collagen type IIB (Col-2B), and aggrecan expression were determined in cartilage. TNFα concentration in the synovial fluid was determined by enzyme-linked immunosorbent assay (ELISA). RESULTS Expression of TNFα, ADAMTS-5 and MMP-13 was significantly increased in synovial tissue from OA joints. Synovial membrane IL-1β abundance showed only moderate elevations in OA, without reaching significant levels. Cytokine expression was increased significantly in OA cartilage samples, particularly TNFα, IL-1β, ADAMTS-4 and MMP-13; and collagen type I expression was significantly increased in synovial tissues from OA groups. Collagen type II message was diminished in mild and moderate stages of OA, but rebounded to significant elevations in severely degenerate joints. Conversely, aggrecan levels significantly declined in cartilage from all OA groups. Synovial fluid TNFα peptide concentration was significantly increased in severe OA cases. CONCLUSION TNFα was increased in all degrees of equine OA, and was abundantly expressed in synovial membrane and cartilage. IL-1β was overexpressed in OA cartilage, but not to a significant extent in synovium. POTENTIAL RELEVANCE Control of TNFα should be considered further as a target in the treatment of OA. ADAMTS-4 may be the primary aggrecanase causing cartilage breakdown in OA.

Effect of speed on stride parameters in racehorses at gallop in field conditions

SUMMARY Stride duration, stance duration and protraction duration are key variables when describing the gaits of terrestrial animals. Together, they determine the duty factor (the fraction of the stride for which the limb maintains contact with the ground surface), from which the peak vertical force can be estimated. When an animal changes speed, these variables change at different proportions. Limited measurements of these variables and predictions of peak limb force have been undertaken for large mammals performing high-speed over-ground exercise. This study set out to make such measurements, employing a previously validated system consisting of limb-mounted accelerometers and a Global Positioning System data logger. Measurements were made on nine elite Thoroughbred racehorses during gallop locomotion over a range of speeds from 9 to 17 m s-1. No statistically significant differences were seen in any variables between the lead and non-lead limbs for either the fore or hind pairs of limbs. Mean stance durations of 131 and 77 ms in the forelimbs and 143 and 94 ms in the hindlimbs were recorded at speeds of 9 and 17 ms-1, respectively. Equivalent values for protraction duration were 364 and 342 (fore) and 355 and 326 ms (hind). Peak limb forces (from duty factor) at 17 ms-1 were 24.7 N kg-1 body weight (range 22.6 to 26.0 N kg-1 body weight) for the forelimbs and 15.3 N kg-1 (range 13.7-16.2 N kg-1 body weight) for the hindlimbs. The duration of the aerial phase of the stride (when no limbs are in contact with the ground) was independent of speed. Overlap time (when more than one leg is on the ground) dropped with speed and approached zero at maximum speed.

Accuracy and precision of hind limb foot contact timings of horses determined using a pelvis-mounted inertial measurement unit

Gait analysis using small sensor units is becoming increasingly popular in the clinical context. In order to segment continuous movement from a defined point of the stride cycle, knowledge about footfall timings is essential. We evaluated the accuracy and precision of foot contact timings of a defined limb determined using an inertial sensor mounted on the pelvis of ten horses during walk and trot at different speeds and in different directions. Foot contact was estimated from vertical velocity events occurring before maximum sensor roll towards the contralateral limb. Foot contact timings matched data from a synchronised hoof mounted accelerometer well when velocity minimum was used for walk (mean (SD) difference of 15 (18)ms across horses) and velocity zero-crossing for trot (mean (SD) difference from -4 (14) to 12 (7)ms depending on the condition). The stride segmentation method also remained robust when applied to movement data of hind limb lame horses. In future, this method may find application in segmenting overground sensor data of various species.

Racing performance after combined prosthetic laryngoplasty and ipsilateral ventriculocordectomy or partial arytenoidectomy: 135 Thoroughbred racehorses competing at less than 2400 m (1997-2007)

REASONS FOR PERFORMING STUDY The success of combined prosthetic laryngoplasty with ipsilateral ventriculocordectomy (LPVC) has not been compared to that of partial arytenoidectomy (PA) in a clinical population. HYPOTHESES In Thoroughbred (TB) racehorses: (1) earnings after LPVC are unaffected by the severity of recurrent laryngeal neuropathy (RLN) (laryngeal grade III vs. grade IV); (2) LPVC and PA yield similar results in the treatment of grade III RLN; (3) performance outcome following PA is independent of diagnosis (RLN vs. unilateral arytenoid chondritis [UAC]); and (4) neither LPVC nor PA returns horses to the level of performance of controls. METHODS Medical and racing records of 135 TB racehorses undergoing LPVC or PA for the treatment of grade III or IV RLN or UAC were reviewed. Racing records of age and sex matched controls were also reviewed. RESULTS After LPVC, horses with grade III RLN performed better compared to those with grade IV RLN. Furthermore, horses treated for grade III RLN by LPVC showed post operative earnings comparable to controls. Rate of return to racing were similar for PA and LPVC, although LPVC resulted in higher post operative earnings. Performance after PA was similar regardless of diagnosis (UAC or RLN). Finally, neither LPVC when performed for grade IV RLN, nor PA performed for either diagnosis restored post operative earnings to control levels. CONCLUSIONS Thoroughbred racehorses treated by LPVC for grade III RLN show significantly better post operative earnings compared to horses treated for grade IV disease. In grade III RLN, LPVC returns earning potential to control levels. PA and LPVC lead to similar success in terms of rate of return to racing, but PA leads to inferior earnings after surgery. POTENTIAL RELEVANCE Laryngoplasty should be recommended for all TB racehorses with grade III RLN to maximise return to racing at a high level. This contradicts the common approach of waiting for complete paralysis.

Equine Laryngoplasty Sutures Undergo Increased Loading During Coughing and Swallowing

OBJECTIVES To report (1) the force required on a single laryngoplasty suture to achieve optimal abduction of the left arytenoid cartilage, (2) peak forces experienced by the suture during induced swallowing and coughing, and during 24-hour resting activity in a stall, and (3) peak forces during induced swallowing and coughing after left recurrent laryngeal nerve blockade. STUDY DESIGN Experimental study. ANIMALS Horses (n=8). METHODS Each laryngoplasty suture was instrumented with an E-type buckle force transducer to measure the force required for optimal intraoperative left arytenoid cartilage abduction. This was correlated with abduction observed postoperatively. Change in suture force from baseline was measured during induced coughing and swallowing, and during normal stall activity. RESULTS Optimal intraoperative arytenoid abduction was achieved with a mean (±SD) force of 27.6±7.5 N. During saline-induced swallowing and coughing mean force on the suture increased by 19.0±5.6 N (n=233 measurements; 7 horses) and 12.1±3.6 N (n=31; 4 horses), respectively. Sutures underwent increased loading a mean of 1152 times in 24 hours. No change in suture force was observed with respiratory rhythm. CONCLUSION Swallowing increases laryngoplasty suture force to a greater extent than coughing.

Intra-articular stabilisation of the equine cricoarytenoid joint.

REASONS FOR PERFORMING STUDY The success of laryngoplasty is limited by abduction loss in the early post operative period. OBJECTIVE To determine the efficacy of polymethylmethacrylate (PMMA) in stabilising the cricoarytenoid joint (CAJ) and reducing the force on the laryngoplasty suture. HYPOTHESIS Injection into the cricoarytenoid joint resists the forces produced by physiological laryngeal air flows and pressures thereby reducing the force experienced by the laryngoplasty suture. METHODS Ten cadaver larynges were collected at necropsy and PMMA was injected into one CAJ at selected random. Each larynx was subjected to physiological conditions with with constant (static) or cycling (dynamic) flow. The specimens were tested sequentially in each of 4 conditions: 1) bilateral full abduction (Control 1); 2) transection of the suture on the side without PMMA; 3) bilateral abduction achieved by replacing the suture (Control 2); and 4) cutting the suture on the PMMA side. Tracheal pressure and flow and pressure in the flow chamber were recorded using pressure and flow transducers. The strain experienced by each suture during bilateral abduction (Controls 1 and 2) was measured. Statistical comparison of the 4 conditions was performed using a mixed effect model with Tukeys post hoc test for multiple comparisons. The strain gauge data were analysed by paired comparison of the regression slopes. RESULTS In the static and dynamic states, tracheal pressure increased and tracheal flow decreased when the suture on the non-cement side was cut (P < 0.05). There was no significant difference in any outcome measure between PMMA injected into the CAJ and bilaterally abducted specimens (Controls 1 and 2) for either condition. The rate of increase in strain with increasing translaryngeal pressure was significantly less on the suture with PMMA placed in the CAJ (P = 0.03). CONCLUSIONS These data provide strong evidence that injecting PMMA into the CAJ resists the collapsing effect of physiological airflows and pressures in vitro and reduces the force experienced by the laryngoplasty suture during maximal abduction. POTENTIAL RELEVANCE Augmentation of prosthetic laryngoplasty with this technique may reduce arytenoid abduction loss in the early post operative period.

Identifying optimal parameters for quantification of changes in pelvic movement symmetry as a response to diagnostic analgesia in the hindlimbs of horses

REASONS FOR PERFORMING STUDY Subjective evaluation of the response to diagnostic analgesia of hindlimb lameness is influenced by expectation bias. Quantification of pelvic movement with inertial measurement units is possible, but it is unclear which measure of movement symmetry best reflects the changes seen after diagnostic analgesia. OBJECTIVES To test our hypothesis that objective measures closely relating to those used for subjective visual lameness scoring (quantifying the difference between sacral upward or downward movement or between movement amplitudes of the left and right tubera coxae) would show the largest and most consistent response. STUDY DESIGN Retrospective analysis of subjective and objective clinical lameness data. METHODS Thirteen horses with hindlimb lameness underwent visual lameness scoring and independent gait assessment with inertial measurement units. Established objective measures were calculated and changes before/after diagnostic analgesia regressed against the change in lameness grade. Slopes of regression lines were calculated and confidence intervals assessed. RESULTS All objective parameters showed variation between horses and across lameness grades. The following 3 measures documented a consistent increase in symmetry for each grade of change in lameness score: difference between sacral displacement minima; upward movement difference; and range of motion difference between the tubera coxae. CONCLUSIONS Several pelvic parameters are suitable for objective quantification of changes after diagnostic analgesia in hindlimb-lame horses. A change of 9-13% per lameness grade can be expected. Upward movement of the tubera coxae is the most sensitive objective measure for quantifying a response to diagnostic analgesia. However, when assessed visually, this measure requires simultaneous focusing on pelvic and limb movements. Other parameters that show a similar response might be more easily perceived, requiring only assessment of tubera coxae range of motion or downward displacement of the sacrum without simultaneous focus on limb movement.