3D Human Neuromuscular Architecture Encoded by Compliant Patterned Hydrogel Scaffolds

Engineered human neuromuscular models are often constrained by a balance between biological accuracy and architectural control. Self-assembled organoids form randomly, while engineered 3D tissues create anisotropy through passive tension at the edges, lacking internal guidance. To address this issue, researchers encode architectural guidance into human neuromuscular tissues by coupling a three-dimensional, cell-laden biomatrix with a xolographically printed hydrogel substrate possessing sub-millimeter surface topography.

They discovered that grooved substrates enhance the alignment and maturation of human primary myotubes within the 3D tissue, while also directing directional axon extension from human iPSC-derived motor neuron spheroids. When these two cell types are aligned along a shared axis, they create functional neuromuscular connections, as evidenced by acetylcholine receptor clustering, glutamate-induced contraction, and its inhibition by d-tubocurarine (curare).

The final construct achieves centimeter-scale innervation coverage, offering a single untethered platform for studying neuromuscular tissue development and function.