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<p>Oligodendrocytes enable rapid central nervous system signaling by myelinating axons. Here, to model key biomechanical cues regulating myelination, we developed a tunable hydrogel-based micropillar array system that mimics the three-dimensional architecture and softness of axons. This platform supports the long-term culture of oligodendrocytes and robust formation of multilayered compact myelin by rodent and human oligodendrocytes. Using confocal and transmission electron microscopy, we observed a strong linear correlation between immunostained myelin thickness and the number of myelin wraps, enabling high-content quantification of myelination. Systematic variation of pillar stiffness, diameter and surface chemistry within pathophysiological ranges revealed that both mechanical and geometric properties of axon-like substrates critically regulate oligodendrocyte differentiation and myelin wrapping. Importantly, we demonstrate that pharmacological agents exhibit stiffness-dependent effects on myelination, suggesting that overly rigid in vitro models may yield false-positive drug hits. This platform offers a physiologically relevant, high-throughput assay for dissecting oligodendrocyte biology and discovering remyelinating therapies for diseases such as multiple sclerosis.</p>

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Tunable hydrogel-based micropillar arrays for myelination studies

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年3月30日

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  • Soufian Lasli,
  • Claire Vinel,
  • Ayushi Agrawal,
  • Yousef Javanmardi,
  • Paola Pedarzani,
  • Beatriz Garcia Diaz,
  • Juan Antonio Garcia-Leon,
  • Boris Djordjevic,
  • Ian J. White,
  • Graham K. Sheridan,
  • William D. Richardson,
  • Emad Moeendarbary

摘要

Oligodendrocytes enable rapid central nervous system signaling by myelinating axons. Here, to model key biomechanical cues regulating myelination, we developed a tunable hydrogel-based micropillar array system that mimics the three-dimensional architecture and softness of axons. This platform supports the long-term culture of oligodendrocytes and robust formation of multilayered compact myelin by rodent and human oligodendrocytes. Using confocal and transmission electron microscopy, we observed a strong linear correlation between immunostained myelin thickness and the number of myelin wraps, enabling high-content quantification of myelination. Systematic variation of pillar stiffness, diameter and surface chemistry within pathophysiological ranges revealed that both mechanical and geometric properties of axon-like substrates critically regulate oligodendrocyte differentiation and myelin wrapping. Importantly, we demonstrate that pharmacological agents exhibit stiffness-dependent effects on myelination, suggesting that overly rigid in vitro models may yield false-positive drug hits. This platform offers a physiologically relevant, high-throughput assay for dissecting oligodendrocyte biology and discovering remyelinating therapies for diseases such as multiple sclerosis.