Clodronate disodium does not produce measurable effects on bone metabolism in an exercising, juvenile, large animal model.

Bisphosphonates are commonly used to treat and prevent bone loss, but their effects in active, juvenile populations are unknown. This study examined the effects of intramuscular clodronate disodium (CLO) on bone turnover, serum bone biomarkers (SBB), bone mineral density (BMD), bone microstructure, biomechanical testing (BT), and cartilage glycosaminoglycan content (GAG) over 165 days. Forty juvenile sheep (253 ± 6 days of age) were divided into four groups: Control (saline), T0 (0.6 mg/kg CLO on day 0), T84 (0.6 mg/kg CLO on day 84), and T0+84 (0.6 mg/kg CLO on days 0 and 84). Sheep were exercised 4 days/week and underwent physical and lameness examinations every 14 days. Blood samples were collected for SBB every 28 days. Microstructure and BMD were calculated from tuber coxae (TC) biopsies (days 84 and 165) and bone healing was assessed by examining the prior biopsy site. BT and GAG were evaluated postmortem. Data, except lameness data, were analyzed using a mixed-effects model; lameness data were analyzed as ordinal data using a cumulative logistic model. CLO did not have any measurable effects on the skeleton of sheep. SBB showed changes over time (p ≤ 0.03), with increases in bone formation and decreases in some bone resorption markers. TC biopsies showed increasing bone volume fraction, trabecular spacing and thickness, and reduced trabecular number on day 165 versus day 84 (p ≤ 0.04). These changes may be attributed to exercise or growth. The absence of a treatment effect may be explained by the lower CLO dose used in large animals compared to humans. Further research is needed to examine whether low doses of bisphosphonates may be used in active juvenile populations for analgesia without evidence of bone changes.

Lumbar vertebral bone density is decreased in horses with pituitary pars intermedia dysfunction.

BACKGROUND: Pathological fractures have been reported in equids with pituitary pars intermedia dysfunction (PPID) but their prevalence and pathogenesis is unknown. OBJECTIVES: To compare: (1) bone mineral density (BMD) in weight bearing and nonweight bearing bones in PPID+ equids and aged and young PPID- controls; and (2) biomechanical properties of the fourth lumbar vertebral body in PPID+ equids and aged PPID- equids. STUDY DESIGN: Case-control study: five PPID+ equids and six aged and four young PPID- control horses. METHODS: PPID status was based on clinical signs and necropsy examination of the pituitary gland (PG). The lumbar vertebral column, right front third metacarpus (MC3), left hind third metatarsus (MT3), and PG were removed after euthanasia. BMD was determined by quantitative computed tomography of regions of interest (ROI) in each bone and biomechanical testing was performed on the fourth lumbar vertebral body. Serum concentrations of parathormone (PTH), ionised Ca++ , 25-hydroxyvitamin D, and osteocalcin (OC) were also measured. Data were analysed using one-way ANOVA and correlation analyses. RESULTS: BMD of trabecular and cortical regions of interest (ROI) of the third, fourth (L4), and fifth lumbar vertebrae were significantly lower in PPID+ equids as compared with aged (p < 0. 001) and young (p < 0.01) PPID- controls. In contrast, no differences were found in BMD of trabecular or cortical ROIs of MC3 and MT3 between groups. No differences were detected in force at fracture, displacement at fracture, Young's modulus or strain of L4 between PPID+ and aged PPID- horses. No differences were found in serum PTH, ionised Ca++ , 25-hydroxyvitamin D, or OC concentrations between groups. MAIN LIMITATIONS: Limited number of equids studied and variation in test results. CONCLUSIONS: BMD of nonweight bearing bones can be decreased with PPID and could increase risk of developing pathological fractures.

Exercising sheep as a preclinical model for musculoskeletal research.

OBJECTIVE: To establish an orthopedic, preclinical, ovine model of controlled exercise using an equine walker. ANIMALS: 20 Dorset-Polypay sheep. PROCEDURES: Sheep underwent 11 weeks of exercise, 4 days per week. Exercise duration and intensity increased until sheep performed 25 minutes at 1.3 m/s and 5 min at 2.0 m/s. Physical/lameness examinations were conducted every 14 days. Blood was collected every 28 days for analysis of serum bone biomarkers (SBB): bone alkaline phosphatase (BALP), procollagen type I amino-terminal propeptide (PINP), carboxy-telopeptide of type I collagen cross-links (CTX-I), tartrate-resistant acid phosphatase 5b (TRAP5b), and receptor activator of nuclear factor-kß ligand (RANKL). RESULTS: Sheep adapted easily to group exercise. Animals grew taller (P = .006) but had a 4% weight loss (P = .003). RANKL was reduced on days 28 and 84 compared to day 56 (P < .05), CTX-1 was reduced on days 28 and 84 compared to days 0 and 56 (P < .05), and TRAP5b was greater on day 28 compared to day 0 (P = .009). BALP and PINP did not change. CLINICAL RELEVANCE: The described preclinical model of exercising sheep has distinct advantages including ease of handling, an established lameness scale, commercially available ovine SBB assays, and the ability to alter footing characteristics and complete circular exercise. Decreasing CTX-I and RANKL with no change in BALP and PINP suggests reduced bone resorption over the study period. Future studies may include a sedentary group or utilize adult animals to alleviate any influence of growth on SBB.

Pharmacokinetics and plasma protein binding of a single dose of clodronate disodium are similar for juvenile sheep and horses.

OBJECTIVE: To determine the single-dose pharmacokinetics of clodronate disodium (CLO) in juvenile sheep and the plasma protein binding (PPB) of CLO in juvenile sheep and horses. ANIMALS: 11 juvenile crossbred sheep (252 ± 6 days) for the pharmacokinetic study. Three juvenile crossbred sheep (281 ± 4 days) and 3 juvenile Quarter Horses (599 ± 25 days) for PPB analysis. METHODS: CLO concentrations were determined using liquid chromatography-mass spectrometry. Pharmacokinetic parameters were calculated by noncompartmental analysis from plasma samples obtained at 0, 0.5, 1, 3, 6, 12, 24, 48, and 72 hours after CLO administered IM at 0.6 mg/kg. PPB was determined using equine and ovine plasma in a single-use rapid equilibrium dialysis system. RESULTS: The mean and range for maximum plasma concentration (Cmax: 5,596; 2,396-8,613 ng/mL), time of maximal concentration (Tmax: 0.5; 0.5-1.0 h), and area under the curve (AUCall: 12,831; 7,590-17,593 h X ng/mL) were similar to those previously reported in horses. PPB in sheep and horses was moderate to high, with unbound fractions of 26.1 ± 5.1% in sheep and 18.7 ± 7.5% in horses, showing less than a 1.4-fold difference. CLINICAL RELEVANCE: The pharmacokinetic parameters and PPB of CLO in juvenile sheep were similar to those previously reported in horses. The results suggest that juvenile sheep can be utilized as an animal model for studying the potential risks and/or benefits of bisphosphonate use in juvenile horses.

Clodronate disodium is neither cytotoxic nor cytoprotective to normal and recombinant equine interleukin-1ß-treated joint tissues in vitro.

OBJECTIVE: To determine the effects of clodronate disodium (CLO) on control and recombinant equine interleukin-1ß (IL-1ß)-treated equine joint tissues. STUDY DESIGN: In vitro experimental study. SAMPLE POPULATION: Cartilage explants, chondrocytes, and synoviocytes (n = 3 horses). METHODS: Monolayer cultures of chondrocytes and synoviocytes from three horses were subjected to: control media (CON), 5 ng/ml CLO (C/low), 50 ng/ml CLO (C/med), 100 ng/ml CLO (C/high), with and without IL-1ß, and 10 ng/ml IL-1ß (IL) alone for 72 hours. Cartilage explants from three horses were subjected to CON, IL, C/low, and C/med with and without IL-1ß for 72 hours. Culture media was analyzed for prostaglandin-E2 (PGE2 ), interleukin-6 (IL-6), and nitric oxide (NO). Explant media was analyzed for glycosaminoglycan (GAG) content and NO. At 72 hours, explant and monolayer culture viability were assessed, and explant GAG content was measured. RESULTS: IL-1ß treatment resulted in higher media concentrations of GAG, NO, PGE2 , and IL-6 compared to the CON treatment (p < .05), demonstrating a catabolic effect of IL-1ß on explants and monolayer cultures. CLO treatments did not increase media concentrations of GAG, NO, PGE2 , or IL-6 compared to CON, indicating no cytotoxic effect. Nevertheless, CLO treatments administered to IL-1ß-treated monolayer cultures and explants did not significantly reduce the inflammatory response regardless of concentration. CONCLUSION: CLO did not demonstrate cytotoxic nor cytoprotective effects in normal and IL-1ß-stimulated chondrocytes, synoviocytes or explants in culture. CLINICAL SIGNIFICANCE: This study does not support the use of CLO as an anti-inflammatory treatment. Further research is necessary to confirm any anti-inflammatory effects of CLO on joint tissues.

Is the Use of Bisphosphonates Putting Horses at Risk? An Osteoclast Perspective.

Osteoclasts are unique and vital bone cells involved in bone turnover. These cells are active throughout the individual's life and play an intricate role in growth and remodeling. However, extra-label bisphosphonate use may impair osteoclast function, which could result in skeletal microdamage and impaired healing without commonly associated pain, affecting bone remodeling, fracture healing, and growth. These effects could be heightened when administered to growing and exercising animals. Bisphosphonates (BPs) are unevenly distributed in the skeleton; blood supply and bone turnover rate determine BPs uptake in bone. Currently, there is a critical gap in scientific knowledge surrounding the biological impacts of BP use in exercising animals under two years old. This may have significant welfare ramifications for growing and exercising equids. Therefore, future research should investigate the effects of these drugs on skeletally immature horses.

Tracking the Impact of Weather on Equine Activity While Pastured.

Keeping horses outdoors on pasture full-time with free access to shelter holds numerous advantages over housing in stalls, promoting both better mental and physical health. One reason for these benefits is the potential for increased physical activity in horses outdoors on pasture versus those confined to stalls. However, it is not guaranteed the horse will take advantage of this opportunity for greater movement. For this reason, it is important to understand the various reasons why horse activity patterns change. The objective of this study was to investigate how various weather factors - including temperature, humidity, precipitation, and wind speed - directly affect equine movement. To achieve this, horses on two similarly-managed farms were equipped with triaxial accelerometers during five independent time periods from January to August. These devices tracked number of steps, standing time, time lying down, and number of lying bouts. The movement data were then compared to the corresponding weather conditions. No strong correlations were found between the recorded movement of the horses and any of the environmental conditions. However, differences in average number of steps and average time lying down were observed between farms and across testing periods, suggesting other influences such as ground conditions and the use of blankets. Further studies are needed to determine the best management practices to encourage pasture activity and support optimal equine physical health.

Average stride length and stride rate of Thoroughbreds and Quarter Horses during racing.

The main factors influencing speed in athletes are stride length (SL) and stride rate (SR). However, conflict remains whether SL or SR is the key determinant of higher speeds. Quarter Horses (QH) generally reach higher speeds in their races than do Thoroughbreds (TB). However, the influence of SL and SR on this greater speed is unclear. Therefore, the main objective of this study was to compare SL and SR in QH and TB raced in short (sprint) and long (classic) distances. We hypothesized that QH have a higher SR in comparison to TB, and SR decreases as distance increases. Two race distances were analyzed for each breed: QH races of 100.6 and 402.3 m, and TB races of 1,207.0 and 2,011.7 m. Data from 20 horses were obtained, consisting of five horses from each race distance (10 QH and 10 TB). Five individuals watched recordings of each race three times counting the number of strides taken by each winning horse. The SR was calculated using the average number of strides over a given race duration, and SL was determined by calculating the total number of strides over the distance covered. Speed was calculated by dividing the distance by the time of the winning horse. The PROC Mixed Procedure was used to identify statistical differences between breeds, and between distances within the same breed. Results showed that although the SL of the TB was longer in comparison with the QH (P < 0.001), the average SR in QH was higher than in TB (2.88 vs. 2.34 + 0.03 strides/s; P < 0.001). Furthermore, QH classic distance demonstrated a faster speed than TB at either distance (P < 0.001). In conclusion, QH achieve a higher SR in comparison to TB (between 14% and 20% more than TB), confirming the importance of SR in achieving high racing speeds.