Relationship between biochemical markers and radiographic scores in the evaluation of the osteoarticular status of Warmblood stallions

Establishing the osteoarticular status of the horse is often performed by means of radiological screening of the animals. Widespread blood sampling could reveal to be an interesting alternative to this procedure which is time consuming and sometimes technically difficult. The aim of this study was to investigate the relationship between the radiological status of the horses and the levels of biochemical markers of cartilage degradation and synovial inflammation. A specific radiological scoring and classification system was therefore developed and applied on 63 stallions presented for studbook admission. Additionally, groups of horses were established according to the occurrence of osteochondrosis, degenerative joint disease and distal interphalangeal joint effusion. Insulin growth factor-I, myeloperoxidases, Coll2-1 and Coll2-1NO₂ were used as blood markers. The combination of the blood parameters did not seem to correlate with the used scoring system. Coll2-1NO₂ levels however tended to increase with poorer radiological class and this could therefore potentially be a useful predictor of the osteoarticular status in the horse. Coll2-1 levels were significantly higher in the degenerative joint disease group. A high percentage of horses with distal interphalangeal joint effusion was present in this study and was associated with decreased IGF-I and increased Coll2-1 levels.     

Method for co-purification of equine neutrophil elastase and myeloperoxidase from a limited blood volume

Neutrophil myeloperoxidase (MPO) and elastase can be released in severe inflammatory diseases and cause tissue injuries. Equine enzymes have already been individually purified from large blood quantities. We describe the isolation of both enzymes from a same limited blood volume. Both MPO and elastase were extracted by crushing PMN isolated by centrifugation on a percoll-gradient from a 460 ml blood collection. MPO and elastase were separated by an ionic exchange chromatography phase and further purified by gel filtration chromatography on Superdex 200 and 75, respectively. Enzymes were identified in the collected fractions by specific enzymatic assays. The final purity was verified by electrophoresis. Specific activity was improved to 19.92 and 34.3× for elastase (final yield: 340 μg) and MPO (final yield: 130 μg), respectively, during the procedure. Results show the possibility of isolating both enzymes from the same blood sample with a sufficient yield and purity for future studies on their implication and interaction during inflammatory diseases.