Changes in biomarkers in equine synovial fluid two weeks after intra-articular hyaluronan treatment: a randomised double-blind clinical trial

Abstract

Background: Inflammatory and degenerative activity inside the joint can be studied in vivo via analysis of synovial fluid (SF) biomarkers, which are molecular markers of inflammatory processes and tissue turnover. The aim of this study was to investigate the response of selected biomarkers in the SF after an intra-articular (IA) high-molecular-weight non-animal stabilized hyaluronic acid (NASHA) treatment. Our hypothesis was that prostaglandin E₂ (PGE₂), substance P, aggrecan chondroitin sulfate 846 epitope (CS846), and carboxypeptide of type II collagen (CPII) concentrations in SF would decrease more in the NASHA than in the placebo group. Twenty-eight clinically lame horses with positive responses to diagnostic IA anaesthesia of the metacarpophalangeal or metatarsophalangeal joints were randomized into treatment (n = 15) and control (n = 13) groups. After collection of baseline SF samples followed by IA diagnostic anaesthesia, horses in the treatment group received 3 ml of a NASHA product IA. Those in the placebo group received an equivalent volume of sterile 0.9% saline solution. The horses were re-evaluated and a second SF sample was obtained after a 2-week period.

Results: CS846 concentration decreased in the NASHA group only (P = 0.010). Both PGE₂ and CPII concentrations decreased within the groups (PGE₂, P = 0.010 for the NASHA group; P = 0.027 for the placebo group; CPII, P < 0.001 for NASHA group; P = 0.009 for placebo group). No significant treatment effect for any biomarker was found between groups. NASHA induced an increase in white blood cell count; this was significant compared with baseline (P = 0.021) and the placebo group (P = 0.045).

Conclusions: Although the SF concentration of the cartilage-derived biomarker CS846 decreased in the NASHA group, no statistically significant treatment effect of any of the biomarkers were observed between treatment groups. The significant increase in SF white blood cell count after IA NASHA may indicate a mild inflammatory response. However, as no clinical adverse effects were observed, we conclude that IA NASHA was well tolerated.

Keywords: Aggrecan chondroitin sulfate 846 epitope; Biomarkers; Carboxypeptide of type II collagen; Clinical study; Non-animal stabilized hyaluronic acid (NASHA); Placebo-controlled; Prostaglandin E2; Substance P.

Fig. 1

Synovial fluid white blood cell count (a), concentrations of total protein (b), prostaglandin E₂ (outliers > 1000 pg/ml are removed to clarify figure) (c), substance P (d), aggrecan chondroitin sulfate 846 epitope (e) and carboxypeptide of type II collagen (f) before and after intra-articular NaCl (placebo) or non-animal stabilized hyaluronic acid (NASHA) injection. First and third quartiles are defined by the boxes and median by the band inside. The 2.5th and the 97.5th percentiles are shown as the lower and upper whiskers. *P < 0.05, **P < 0.001 between first and second sample in NaCl (placebo) and NASHA group respectively, † P < 0.05 NaCl (placebo) and NASHA groups