NASC advances equine joint supplement substantiation using CytoSolve®’s systems architecture for in silico combination screening across four core pathways

National Animal Supplement Council (NASC)
The National Animal Supplement Council is a nonprofit trade association focused on responsible animal supplement use through scientific substantiation, quality standards, and industry accountability. To strengthen evidence-based evaluation of multi-ingredient equine joint formulations, NASC engaged CytoSolve®’s Product Testing Division to perform an independent, mechanistically grounded in silico assessment of combination efficacy.

Challenge

Equine joint health is shaped by multiple interconnected processes—inflammation, oxidative stress, cartilage degeneration, and cartilage regeneration—that evolve together over time. Yet joint supplements are typically multi-ingredient combinations, making it difficult to isolate and quantify:

Single-ingredient vs. synergistic effects across molecular pathways

Cross-talk and interdependence among biological systems (e.g., inflammation amplifying oxidative stress and degeneration)

Time-dependent outcomes that are costly and complex to measure experimentally with sufficient control and repeatability

NASC required a rigorous systems-level approach that could evaluate whether combination formulations provide measurable synergy beyond individual ingredient effects while maintaining mechanistic traceability suitable for scientific substantiation.

How CytoSolve Helped

CytoSolve® applied its computational systems biology platform to execute a controlled, repeatable in silico combination screening of an Equine Joint Formula containing chondroitin sulfate, glucosamine, sodium hyaluronate, methylsulfonylmethane (MSM), and vitamin C.

Key elements of the systems architecture and workflow included:

Systematic literature review to define the pathway blueprint governing equine joint health mechanisms

Translation into individually validated mathematical models representing four major biological systems:

  • Inflammation: PGE2 dynamics via arachidonic acid metabolism
  • Oxidative stress: reactive oxygen species (ROS) generation and modulation
  • Cartilage degeneration: MMP-13 expression as a degradative mediator
  • Cartilage regeneration: collagen-II synthesis as a regenerative readout
Dynamic integration of validated pathway models within CytoSolve® to preserve molecular interdependencies and pathway cross-talk

Dose-relevant simulation design: modeling individual ingredient effects at recommended dose levels

Combination screening simulations: quantifying synergy for multi-ingredient formulations over a 30-day simulated period

Controlled comparisons: direct, repeatable evaluation of single-ingredient vs. combination performance within one unified computational framework

Key Benefits Realized

  • Identified four core molecular systems governing equine joint health in a coherent mechanistic framework
  • Enabled quantitative comparison of individual ingredient effects versus combination outcomes
  • Demonstrated synergistic reductions in inflammation and oxidative stress biomarkers (PGE2, ROS)
  • Showed coordinated downregulation of cartilage degeneration signaling via reduced MMP-13 expression
  • Indicated enhanced cartilage regeneration, reflected by increased collagen-II synthesis under combination conditions
  • Delivered an independent, objective assessment aligned with industry-wide expectations for scientific substantiation

Outcome

CytoSolve®’s in silico combination screening showed that the Equine Joint Formula ingredients act synergistically, producing substantially stronger improvements in joint health biomarkers than individual ingredients alone. Across the 30-day simulations, the combination produced 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.

For NASC, the study provided a mechanistically validated, systems-level assessment of combination efficacy—reinforcing that multi-ingredient equine joint supplements should be evaluated as integrated biological systems rather than isolated components, and positioning CytoSolve®’s platform as a scalable approach for combination screening and rigorous scientific substantiation in animal health supplements.