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Synovial Fluid Metabolites Differentiate between Septic and Nonseptic Joint Pathologies

ABSTRACT: Osteoarthritis (OA), osteochondrosis (OC), and synovial sepsis in horses cause loss of function and pain. Reliable biomarkers are required to achieve accurate and rapid diagnosis, with synovial fluid (SF) holding a unique source of biochemical information. Nuclear magnetic resonance (NMR) spectroscopy allows global metabolite analysis of a small volume of SF, with minimal sample preprocessing using a noninvasive and nondestructive method. Equine SF metabolic profiles from both nonseptic joints (OA and OC) and septic joints were analyzed using 1D 1H NMR spectroscopy. Univariate and multivariate statistical analyses were used to identify differential metabolite abundance between groups. Metabolites were annotated via 1H NMR using 1D NMR identification software Chenomx, with identities confirmed using 1D 1H and 2D 1H 13C NMR. Multivariate analysis identified separation between septic and nonseptic groups. Acetate, alanine, citrate, creatine phosphate, creatinine, glucose, glutamate, glutamine, glycine, phenylalanine, pyruvate, and valine were higher in the nonseptic group, while glycylproline was higher in sepsis. Multivariate separation was primarily driven by glucose; however, partial-least-squares discriminant analysis plots with glucose excluded demonstrated the remaining metabolites were still able to discriminate the groups. This study demonstrates that a panel of synovial metabolites can distinguish between septic and nonseptic equine SF, with glucose the principal discriminator.

INSTRUMENT(S): Bruker

SUBMITTER: Marie Margaret Phelan

PROVIDER: MTBLS543 | MetaboLights | 2018-07-20

REPOSITORIES: MetaboLights

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Project description:<h4><strong>BACKGROUND: </strong>Palmar osteochondral disease (POD) is a common cause of lameness in competition horses. Magnetic resonance imaging (MRI) is the most sensitive diagnostic modality currently available, however it may not be financially or logistically practical for routine screening of POD. There is increasing interest in the use of metabolomics for diagnosis prior to progression to irreversible damage.</h4><h4><strong>OBJECTIVES: </strong>To determine metabolite levels in synovial fluid (SF) of horses with a clinical diagnosis of POD based on diagnostic analgesia and MRI, with the hypothesis that metabolomic profiles differ between diseased and healthy joints.</h4><h4><strong>STUDY DESIGN:</strong> Prospective clinical study.</h4><h4><strong>METHODS: </strong>Synovial fluid was collected from metacarpo/tarsophalangeal joints (MC/TPJ) of 29 horses (n = 51 joints), including 14 controls (n = 26) and 15 cases (n = 25), the latter with lameness localised to the MC/TPJ and MR changes consistent with POD (n = 23). Spectra were produced using <sup>1</sup> H-nuclear magnetic resonance (NMR) spectroscopy and analysed.</h4><h4><strong>RESULTS: </strong>Twenty-five metabolites were recognised associated with various biosynthetic and degradation pathways. The metabolite abundances within the controls demonstrated increased variability compared with the clinical group. The low level of variance between the spectra of the two groups was explained by five principal components. Cross-validation of the cohort demonstrated modest separation of predictive power (R<sup>2</sup> = 0.67; Q<sup>2</sup> = 0.34). Although statistical significance was not achieved, the most influential metabolites were glucose and lactate.</h4><h4><strong>MAIN LIMITATIONS:</strong> The modest sample size and variation in signalment, background and presenting condition of the controls may have impacted the discriminative power of the constructed models. The lack of matched controls, differences in time of fluid collection and freezing times may have also reduced accuracy when representing metabolite profiles.</h4><h4><strong>CONCLUSIONS:</strong> This study identified and quantified metabolites present in MC/TPJ SF of clinical cases with POD.</h4>

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Synovial Fluid Metabolites Differentiate between Septic and Nonseptic Joint Pathologies

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