Engineering Sensory Ganglion Multicellular System to Model Tissue Nerve Ingrowth

Discogenic pain is associated with deep nerve ingrowth in annulus fibrosus tissue (AF) of intervertebral disc (IVD). To model AF nerve ingrowth, primary bovine dorsal root ganglion (DRG) micro-scale tissue units are spatially organised around an AF explant by mild hydrodynamic forces within a collagen matrix. This results in a densely packed multicellular system mimicking the native DRG tissue morphology and a controlled AF-neuron distance. Such a multicellular organisation is essential to evolve populational-level cellular functions and in vivo-like morphologies. Pro-inflammatory cytokine-primed AF demonstrates its neurotrophic and neurotropic effects on nociceptor axons. Both effects are dependent on the AF-neuron distance underpinning the role of recapitulating inter-tissue/organ anatomical proximity when investigating their crosstalk. This is the first in vitro model studying AF nerve ingrowth by engineering mature and large animal tissues in a morphologically and physiologically relevant environment. The new approach can be used to biofabricate multi-tissue/organ models for untangling pathophysiological conditions and develop novel therapies.This is the first in vitro annulus fibrosus (AF) nerve ingrowth model where the AF's neurotrophic and neurotropic effects are demonstrated in a mature large animal dorsal root ganglion (DRG) system. Acoustic generated mild hydrodynamic forces orchestrate physiological-relevant multicellular DRG morphology and anatomical-relevant AF-DRG distance, which determines their evolved functions.image