In silico Combination Screening for Animal Health — CytoSolve’s Equine Joint Health Analysis for NASC

The National Animal Supplement Council (NASC) is a nonprofit trade association dedicated to advancing the responsible use of animal supplements through scientific substantiation and quality standards. To support evidence-based evaluation of multi-ingredient equine joint formulations, NASC commissioned CytoSolve’s Product Testing Division to conduct an independent, in silico combination screening analysis focused on equine joint health mechanisms.

Challenge

Equine joint health is governed by multiple interconnected physiological processes, including inflammation, oxidative stress, cartilage degeneration, and cartilage regeneration. While equine joint supplements often combine several bioactive ingredients, the individual versus synergistic effects of these ingredients across molecular pathways are difficult to isolate using traditional in vivo or in vitro approaches. NASC required a rigorous, systems-level method to evaluate whether combination formulations deliver synergistic efficacy beyond single-ingredient effects, while maintaining biological relevance and mechanistic clarity.

How CytoSolve Helped

CytoSolve applied its patented computational systems biology platform to perform a combination screening analysis of an Equine Joint Formula containing chondroitin sulfate, glucosamine, sodium hyaluronate, methylsulfonylmethane (MSM), and vitamin C. The methodology included:

  • Conducting a systematic literature review to identify molecular pathways governing equine joint health
  • Translating four major biological systems into individually validated mathematical models:
    • Inflammation (PGE2 via arachidonic acid metabolism)
    • Oxidative stress (reactive oxygen species, ROS)
    • Cartilage degeneration (MMP-13 expression)
    • Cartilage regeneration (collagen-II synthesis)
  • Integrating validated pathway models within the CytoSolve platform to preserve molecular interdependencies
  • Simulating individual ingredient effects at recommended dose levels
  • Performing combination screening simulations to quantify synergistic effects of multi-ingredient formulations over a 30-day period

This approach enabled controlled, repeatable comparison of single-ingredient and combination outcomes within a unified biological framework.

Key Benefits Realized

  • Identification of four core molecular systems governing equine joint health
  • Quantitative comparison of individual versus combination ingredient effects
  • Demonstration of synergistic reductions in inflammation and oxidative stress biomarkers
  • Evidence of coordinated downregulation of cartilage degeneration pathways
  • Evidence of enhanced cartilage regeneration through combination effects
  • Independent, objective validation suitable for industry-wide scientific standards

Outcome

CytoSolve’s in silico combination screening demonstrated that the Equine Joint Formula ingredients act synergistically, producing significantly greater improvements in joint health biomarkers than individual ingredients alone. Combination simulations showed marked reductions in inflammatory mediators (PGE2), substantial suppression of oxidative stress (ROS), downregulation of cartilage-degrading enzymes (MMP-13), and increased collagen-II production supporting cartilage regeneration.

This study provided NASC with a mechanistically validated, systems-level assessment of combination efficacy in equine joint supplements. The results underscore the importance of evaluating multi-ingredient formulations as integrated biological systems rather than isolated components, and highlight CytoSolve’s platform as a powerful tool for combination screening and scientific substantiation in animal health supplements.