Cartilage-Sparing Properties of EOC®
Abstract
This study aimed to determine the anti-inflammatory and/or chondroprotective effects of Equine Omega Complete® (EOC®) on cartilage explants stimulated with lipopolysaccharide (LPS).
Explants were aseptically prepared from the intercarpal joints of 17 market-weight pigs and placed in culture at 37°C for a total of 120 hours. For the final 96 hours, explants were conditioned with a simulated digestion extract of EOC® (0, 36 or 180 μL/mL), and for the final 48 hours, explants were stimulated with LPS (0 or 15µg/mL).
Media was removed and replaced every 24 hours. Samples from the final 48 hours were analyzed for biomarkers of cartilage inflammation [prostaglandin E2 (PGE2) and nitric oxide (NO)] and cartilage structure [glycosaminoglycan (GAG)].
At the end of the culture period, cartilage explants were stained for an estimate of cell viability. Stimulation of unconditioned explants with LPS significantly increased media concentrations of PGE2, GAG and NO compared with that from unstimulated explants. LPS stimulation did not significantly affect cell viability.
Both concentrations of EOC® prevented significant LPS-stimulated cartilage release of GAG without impairing chondrocyte viability. No other effects of treatment were observed.
These data provide evidence for EOC®’s non-cytotoxic, chondroprotective effect in cartilage. This in vitro experiment supports the use of EOC in protecting against the detrimental effects of inflammation on cartilage structure.
Equine Omega Complete® is an all-in-one prevention supplement for horses. Targeting joint, gut, respiratory, reproduction muscle and skin and coat health in the horse, the supplement has received proven results in a recent joint study at the University of Guelph, Department of Animal Biosciences.
The research results from their recent trial at the University of Guelph, entitled “Cartilage-sparing properties of Equine Omega Complete® (EOC®) in an organ culture model of cartilage inflammation,” was conducted by Dr. Wendy Pearson, Ph.D.; student Anna Garland, and undergraduate student Corina Wierenga, in the Department of Animal Biosciences.
The study was designed to determine the ability of EOC® to protect cartilage from the damaging effects of inflammation. The researchers pre-treated live cartilage pieces with EOC®, while some were left untreated, and then all the cultures were stimulated to ‘behave’ arthritic.
In cartilage pieces that were not pre-treated with EOC®, the stimulus caused the breakdown of cartilage structure, which was measured by the loss of an important building block of cartilage (called ‘GAG’) from the cartilage pieces. This loss of cartilage building blocks was prevented by pre-treating the cartilage with EOC® at a dose approximating 60 and 180mL per day for a 500kg horse.
Furthermore, exposing cartilage to EOC® at these doses was safe for the cells within the cartilage pieces.
The study provides evidence for the ability of EOC® to protect the cartilage structure of horses at risk for joint inflammation and/or arthritis and supports the use of the product to preserve the health of joints.
Figure 1: GAG loss from cartilage between 0 and 48 h of culture with an inflammatory stimulus. Loss of GAG indicates damage to cartilage structure; this loss was lower when cartilage was exposed to EOC at 1 and 3 times the manufacturer recommended dose.
Figure 2: Small discs of cartilage are removed from the joint surface (A) and placed into a nutrient broth to keep the cartilage pieces alive (B).
Figure 3: GAG analysis is conducted by placing a small volume of the nutrient broth pictured in 2B above and applying a blue stain which attaches to the GAG molecules present in the broth. When the stain attaches, the blue colour changes to pink – the more GAG present, the more pink the sample looks. An instrument called a spectrophotometer reads the change in colour, and the absorbance from the spectrophotometer is converted to a concentration of GAG in the sample.