Purpose
To quantify direct effects of Hyalcare (HC) on cartilage inflammation in vitro
Background
Non-steroidal anti-inflammatory drugs are frequently administered to horses with articular pain and inflammation. There is evidence that these drugs, whilst being effective pain killers, can interfere with the ability of cartilage to launch a repair response in the face of an inflammatory challenge. This may have important implications to the development of degenerative joint disease in horses.
Methods
Cartilage explants (4mm pieces) were removed aseptically from the intercarpal joint of 6 pigs after humane slaughter in a federally inspected meat processing facility. Explants were placed into a 24-well tissue culture plate for 24 h before simulated digest of Hyalcare (3 doses – HC1, HC2 and HC3) was added to the media. One group of control explants was cultured without any HC . Explants were cultured with their treatments for a total duration of 96 hours, the final 48 h of which they were stimulated with a pro-inflammatory molecule (IL-1). Media was removed and refreshed every 24 hours, and was analyzed for compounds involved in pain signaling [PGE2 (ELISA) and NO (Griess Reaction)], and cartilage structure [GAG (dimethyl methylene blue assay).
Results
Addition of IL-1 to control explants increased media PGE2, GAG and NO. HC prevented IL-1-induced increase in NO and GAG, while increasing PGE2.
Fig 1. Media nitrite (A) and GAG (B) from explants stimulated with IL-1 in the presence or absence of hyalcare (HCsim). * denotes significant effect of IL-1 within treatments; lower-case letters denote significant differences between treatments at a single time point.
Conclusions
These data provide evidence that Hyalcare protects against loss of cartilage structure in the face of an inflammatory challenge. Furthermore the product reduced production of nitric oxide (a molecule involved in pain signaling) which supports the use of Hyalcare to reduce pain.