Posterolateral spinal fusion (PLF) is a procedure used for the treatment of degenerative spine disease. In this study we evaluated Osteogrow-C, a novel osteoinductive device comprised of recombinant ...human Bone morphogenetic protein 6 (rhBMP6) dispersed in autologous blood coagulum with synthetic ceramic particles, in the sheep PLF model. Osteogrow-C implants containing 74-420 or 1000-1700 µm ceramic particles (TCP/HA 80/20) were implanted between L4-L5 transverse processes in sheep (Ovis Aries, Merinolaandschaf breed). In the first experiment (n = 9 sheep; rhBMP6 dose 800 µg) the follow-up period was 27 weeks while in the second experiment (n = 12 sheep; rhBMP6 dose 500 µg) spinal fusion was assessed by in vivo CT after 9 weeks and at the end of the experiment after 14 (n = 6 sheep) and 40 (n = 6 sheep) weeks. Methods of evaluation included microCT, histological analyses and biomechanical testing. Osteogrow-C implants containing both 74-420 and 1000-1700 µm ceramic particles induced radiographic solid fusion 9 weeks following implantation. Ex-vivo microCT and histological analyses revealed complete osseointegration of newly formed bone with adjacent transverse processes. Biomechanical testing confirmed that fusion between transverse processes was complete and successful. Osteogrow-C implants induced spinal fusion in sheep PLF model and therefore represent a novel therapeutic solution for patients with degenerative disc disease.
Abstract
Autologous bone graft substitute (ABGS) containing rhBMP6 in autologous blood coagulum (Osteogrow) is a novel therapeutic solution for bone regeneration. This study is aimed to investigate ...the long-term outcome of ABGS with synthetic ceramics (Osteogrow-C) in rabbit posterolateral spinal fusion (PLF) model. Osteogrow-C implants were implanted bilaterally between rabbit lumbar transverse processes. We compared the outcome following implantation of ABGS with ceramic particles of different chemical composition (TCP and biphasic ceramics containing both TCP and HA) and size (500–1700 µm and 74–420 µm). Outcome was analyzed after 14 and 27 weeks by microCT, histology, and biomechanical analyses. Successful bilateral spinal fusion was observed in all animals at the end of observation period. Chemical composition of ceramic particles has impact on the PLF outcome via resorption of TCP ceramics, while ceramics containing HA were only partially resorbed. Moreover, persistence of ceramic particles subsequently resulted with an increased bone volume in implants with small particles containing high proportion of HA. ABGS (rhBMP6/ABC) with various synthetic ceramic particles promoted spinal fusion in rabbits. This is the first presentation of BMP-mediated ectopic bone formation in rabbit PLF model with radiological, histological, and biomechanical features over a time course of up to 27 weeks.
In human and veterinary medicine calprotectin is most widely used in diagnosing different gastro-intestinal diseases. The aim of this study was to assess the stability of canine calprotectin (cCP) in ...serum after storage at low temperatures and imprecision of the method.
Blood samples were collected from dogs with different clinical diagnoses. Twenty-two dogs were included in this study. Calprotectin concentration was measured 4 hours after serum separation (T0), and after being frozen at - 80 °C for 8 (T1) and 16 weeks (T2). The maximum permissible difference (MPD) was derived from the equation for calculating total error (TE) TE = %Bias + (1.96 x %CV), where bias and coefficient of variation (CV) were defined by the manufacturer. The dogs enrolled in this study were patients admitted during the morning (9-12 a.m.), on the day the first measurement was performed. All sample analysis for determination of stability were done in duplicates. For determination of within-run precision, the two patients' serum samples were analyzed in 20 replicates. Imprecision was assessed by analyzing 20 replicates on one plate on two samples where high and low concentrations were anticipated.
The calculated value of MPD was 32.52%. Median calprotectin concentrations were higher at T1 114.08 μg/L (IQR = 55.05-254.56) and T2 133.6 μg/L (IQR = 100.57-332.98) than at T0 83.60 μg/L (IQR = 50.38-176.07). Relative and absolute bias at T1 (49.3%; 45.98 μg/L) and T2 (109.93%; 94.09 μg /L) have shown that calprotectin concentrations increase after long term storage at - 80 °C.
The results of the present study indicate that c-CP was not stable for 16 weeks at low storage temperature (- 80 °C). Considering the observed change in the concentration of c-CP at T1, a storage time of 8 weeks should be safely applied. The method imprecision was not satisfactory, especially in the lower concentration range.
Over a year into the COVID-19 pandemic, there is growing evidence that SARS-CoV-2 infections among dogs are more common than previously thought. In this study, the prevalence of SARS-CoV-2 antibodies ...was investigated in two dog populations. The first group was comprised of 1069 dogs admitted to the Veterinary Teaching Hospital for any given reason. The second group included dogs that shared households with confirmed COVID-19 cases in humans. This study group numbered 78 dogs. In COVID-19 infected households, 43.9% tested ELISA positive, and neutralising antibodies were detected in 25.64% of dogs. Those data are comparable with the secondary attack rate in the human population. With 14.69% of dogs in the general population testing ELISA positive, there was a surge of SARS-CoV-2 infections within the dog population amid the second wave of the pandemic. Noticeably seroprevalence of SARS-CoV-2 in the dog and the human population did not differ at the end of the study period. Male sex, breed and age were identified as significant risk factors. This study gives strong evidence that while acute dog infections are mostly asymptomatic, they can pose a significant risk to dog health. Due to the retrospective nature of this study, samples for viral isolation and PCR were unavailable. Still, seropositive dogs had a 1.97 times greater risk for developing central nervous symptoms.
A special part of archaeology, so-called archaeozoopathology or veterinary paleopathology is dedicated to studies of paleopathological changes in animal remains and contributes to the knowledge of ...ancient veterinary medicine and the history of diseases. In our study, we analyze paleopathological changes determined by gross observation and diagnostic imaging in the animal material originating from eight archaeological sites in Croatia. A standard archaeozoological analysis was carried out and specimens with visually detected macrostructural changes were radiographed. In total, 50 animal remains with altered macrostructure were identified in the archaeozoological material excavated from 2010 to 2022 at eight archaeological sites in Croatia. According to the taxonomic analysis, most of the bones with macrostructural changes originated from cattle (N = 27, 54% of the total number of bones with macrostructural changes), followed by the bones of small ruminants (N = 12, 24%) and pigs (N = 8, 16%). The horse, carnivore and chicken were represented with one bone each (2%). Radiological examination showed that three samples (6%) had a regular bone macrostructure, i.e., no pathological changes were visible upon radiological examination. The majority (64%) of pathologically altered bones are a consequence of keeping/working, followed by traumatic causes (20%). Changes in the oral cavity were found in 10% of specimens. Our study showed that gross examination will continue to be the primary method for the identification of pathologically altered remains in archaeozoological material. However, diagnostic imaging techniques such as radiography should be implemented to confirm or exclude suspected alterations and to help the classification of the specimen by etiology.
This study aimed to evaluate if exercise-induced acute phase response (APR) occurs in endurance horses in response to the race. The study included 23 horses competing in an endurance competition with ...a successfully passed clinical examination before the race. Blood samples were collected before the start and within 30 min after the end of the race. Haematological and biochemical tests were performed and correlated to acute phase biomarkers changes. Values of calprotectin and haptoglobin (Hp) decreased after the races compared to values before, while concentrations of ceruloplasmin and albumin recorded a significant increase. Greater changes in calprotectin values were noted in Arabian horses compared to other breeds. Values of Hp showed a significantly greater decrease after longer races. Based on study results, endurance racing induces APR in horses characterised by significant changes in selected acute phase biomarkers. More pronounced changes were noted at races with higher average speeds, suggesting the need for thorough horse monitoring during exhausting races.
Oxidative stress (OS) associated with an intense exercise may have a negative influence on equine health. The aim of this study was to determine the effects of endurance races on oxidative and ...antioxidative status of horses by evaluating changes in reactive oxygen metabolites (d-ROMs), malondialdehyde (MDA), biological antioxidant potential (BAP) and oxidative stress index (OSI) values. The study was carried out on 53 race starts (28 individual horses) competing at different endurance races according to distance (40 and 80 km) and difficulty (easy and demanding). Blood samples were taken before and after the race.
Compared to levels of OS serum biomarkers before the race, an increase in values of d-ROMs (P < 0.01), MDA (P < 0.01), and BAP (P < 0.001), and a decrease in OSI (P < 0.001) have been noted after the race. Contrary to other measured biomarkers, BAP did not show significant individual effects of horses. Horses competing at shorter races have shown a significant change in d-ROMs (P = 0.002), BAP (P < 0.001) and OSI (P = 0.004), whereas those competing at longer races in MDA (P = 0.002), BAP (P < 0.001) and OSI (P < 0.001) post-race values. Endurance racing induced changes in values of d-ROMs, BAP and OSI during both easy and demanding races.
Changes in all measured OS biomarkers indicate that prolonged aerobic exercise during endurance race could contribute to the imbalance between oxidants and antioxidants in horses, mainly characterised by a pronounced antioxidant response. Biological antioxidant potential was found to be the most reliable biomarker of OS in endurance horses in the present study.
Management of large segmental defects is one of the most challenging issues in bone repair biology. Autologous bone graft substitute (ABGS) containing rhBMP6 within autologous blood coagulum (ABC) ...with synthetic ceramics is a novel biocompatible therapeutic solution for bone regeneration.
A 2-year old dog was brought to the veterinary clinics due to pain and bleeding from the right front leg after being unintendedly hit by a gunshot. Radiological examination revealed a large, 3 cm long multisegmental defect of the humerus on the right front leg with a loss of anatomical structure in the distal portion of the bone. The defect was treated surgically and an external fixator was inserted to ensure immobilization. Complete lack of bone formation 3 months following surgery required a full reconstruction of the defect site with a novel ABGS (rhBMP6 in ABC with ceramic particles) to avoid front leg amputation. The healing was then followed for the next 16 months. The callus formation was observed on x-ray images 2 months following ABGS implantation. The bone segments progressively fused together leading to the defect rebridgment allowing removal of the external fixator by 4 months after the reconstruction surgery. At the end of the observation period, the function of the leg was almost fully restored while analyses of the humeral CT sections revealed restoration and cortices rebridgment with a renewal of uniform medullary canal including structural reconstruction of the distal humerus.
This large humeral gunshot segmental defect of the front leg in a dog was saved from amputation via inducing bone regeneration using a novel ABGS osteoinductive device containing BMP6 in ABC.
The beneficial effect of physical activity on the musculoskeletal health in dogs is well recognized, but the level of intensity, duration, and frequency of exercise is not fully described. ...Measurement of serum markers of bone metabolism (bone alkaline phosphatase and osteocalcin as bone formation markers and C-terminal telopeptide as bone resorption marker) during four months of organized moderate-intensity physical training in Labrador retriever and Golden retriever dogs aged between 11.7–24.4 months, showed variations of bone metabolism. Dogs were included in treadmill running sessions for 25 min, three times per week. Blood samples were taken at the beginning of the program (baseline), after two months (mid-term) and at the end of the study after four months. The values of bone alkaline phosphatase and osteocalcin significantly decreased following two months of exercise program. Bone alkaline phosphatase increased by the end of four-month training cycle, but did not reach baseline value. Osteocalcin levels continued to decrease towards the end of the study. C-terminal telopeptide concentrations did not significantly change throughout the study duration. The results of this study show that aerobic exercise of moderate-intensity caused an initial decrease in bone formation followed by an increase of bone alkaline phosphatase and a further decrease of osteocalcin concentration. The response of two formation markers can be explained by the different stage of osteoblast activity that they express. In summary, moderate exercise resulted in no change in bone resorption, and a mild bone formation in young developing dogs.
During intensive physical activity horses are exposed to thermolysis, electrolyte loss and rising amounts of catabolic products, which results in alterations of biochemical blood parameters due to ...the horse’s adaptation to metabolic stress. The aim of the present study was to determine the effect of intensive physical activity on serum biochemical parameters in horses competing in 40 and 80 km endurance races. Blood samples were taken from 28 horses before and after the race over four competitions, with a total of 53 samples analyzed. Biochemical parameters studied included creatine kinase, lactate dehydrogenase, aspartate aminotransferase, urea, creatinine, gamma-glutamyl transferase, glucose, triglycerides, cholesterol, sodium, potassium, calcium and magnesium. The extent of change in pre- and post-race values was compared between categories of age, gender, breed, distance and average speed of horses. Creatine kinase and blood glucose values prior to the race were higher than the reference values. Values of renal parameters and parameters of muscle damage increased after the races, and the degree of change was more pronounced at longer racing lengths, as well as at lower average speed. Electrolyte loss was more prominent during longer races. This study demonstrated that endurance races cause evident changes in serum electrolyte concentrations, renal parameters and markers of muscle damage in horses. These changes are observable in horses at both short and long duration endurance races.
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