Impact of Long-Term Storage on Mid-Infrared Spectral Patterns of Serum and Synovial Fluid of Dogs with Osteoarthritis

Mid-infrared spectral (MIR) patterns of serum and synovial fluid (SF) are candidate biomarkers of osteoarthritis (OA). The impact of long-term storage on MIR spectral patterns was previously unknown. MIR spectra of canine serum (52 knee-OA, 49 control) and SF (51 knee-OA, 51 control) were obtained after short-term and long-term storage in −80 °C. Multilevel simultaneous component analysis and partial least squares discriminant analysis were used to evaluate the effect of time and compare the performance of predictive models for discriminating OA from controls. The median interval of storage between sample measurements was 5.7 years. Spectra obtained at two time points were significantly different (p < 0.0001); however, sample aging accounted for only 1.61% and 2.98% of the serum and SF profiles’ variability, respectively. Predictive models for discriminating serum of OA from controls for short-term storage showed 87.3 ± 3.7% sensitivity, 88.9 ± 2.4% specificity, and 88.1 ± 2.3% accuracy, while for long-term storage, they were 92.5 ± 2.6%, 97.1 ± 1.7%, and 94.8 ± 1.4%, respectively. Predictive models of short-term stored SF spectra had 97.3 ± 1.6% sensitivity, 89.4 ± 2.6% specificity, and 93.4 ± 1.6% accuracy, while for long-term storage they were 95.7 ± 2.1%, 95.7 ± 0.8%, and 95.8 ± 1.1%, respectively. Long-term storage of serum and SF resulted in significant differences in MIR spectral variables without significantly altering the performance of predictive algorithms for discriminating OA from controls.