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<p>Exosomes mediate signaling that guides differentiation by delivering factors that influence cellular fate and gene pathways. Three-dimensional (3D)–printed scaffolds provide customizable matrices that create precise microenvironments for differentiation and can synergize with exosome cues to enhance lineage specification and tissue regeneration. This study evaluated the neurogenic differentiation potential of exosomes derived from neural stem cells (NSCs-exs) on adipose-derived mesenchymal stem cells (ADMSCs) cultured on a polyacrylonitrile (PAN) scaffold prepared by 3D printing. NSCs-exs were characterized by transmission electron microscopy (TEM), Western blotting, and dynamic light scattering (DLS). The PAN scaffold was fabricated by a solution-printing method using a 3D printer to deposit a 10% PAN polymer mixture. ADMSCs were cultured on the scaffold and treated with 10&#xa0;µg/mL NSCs-exs for 14 days. Differentiation was assessed by examining the expression of Nestin, Map2, Tuj-1, and NF using immunocytochemistry and RT-PCR. NSCs-exs were characterized by a diameter of 44.79&#xa0;nm, a lipid bilayer structure, and expression of the markers CD81, CD9, and CD63, with a total protein concentration of 160&#xa0;µg/mL. Microstructural analysis of the PAN scaffold revealed a homogeneous and organized architecture with an average fiber diameter of approximately 470&#xa0;nm. The scaffold exhibited appropriate chemical bonding, sufficient mechanical strength, and biocompatibility. A significant increase (<i>P</i> ≤ 0.05) in neural-marker expression indicated successful neural differentiation of ADMSCs under NSCs-exs treatment in 3D culture. Exosome-guided differentiation on 3D PAN scaffolds promotes neural lineage commitment, reinforcing exosomes as reparative tools for neural regeneration.</p>

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Fabrication and characterization of 3D-printed polyacrylonitrile scaffolds for the neural differentiation of mesenchymal stem cells via exosomes

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年4月3日

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  • Elham Hoveizi,
  • Mostafa Sayahi

摘要

Exosomes mediate signaling that guides differentiation by delivering factors that influence cellular fate and gene pathways. Three-dimensional (3D)–printed scaffolds provide customizable matrices that create precise microenvironments for differentiation and can synergize with exosome cues to enhance lineage specification and tissue regeneration. This study evaluated the neurogenic differentiation potential of exosomes derived from neural stem cells (NSCs-exs) on adipose-derived mesenchymal stem cells (ADMSCs) cultured on a polyacrylonitrile (PAN) scaffold prepared by 3D printing. NSCs-exs were characterized by transmission electron microscopy (TEM), Western blotting, and dynamic light scattering (DLS). The PAN scaffold was fabricated by a solution-printing method using a 3D printer to deposit a 10% PAN polymer mixture. ADMSCs were cultured on the scaffold and treated with 10 µg/mL NSCs-exs for 14 days. Differentiation was assessed by examining the expression of Nestin, Map2, Tuj-1, and NF using immunocytochemistry and RT-PCR. NSCs-exs were characterized by a diameter of 44.79 nm, a lipid bilayer structure, and expression of the markers CD81, CD9, and CD63, with a total protein concentration of 160 µg/mL. Microstructural analysis of the PAN scaffold revealed a homogeneous and organized architecture with an average fiber diameter of approximately 470 nm. The scaffold exhibited appropriate chemical bonding, sufficient mechanical strength, and biocompatibility. A significant increase (P ≤ 0.05) in neural-marker expression indicated successful neural differentiation of ADMSCs under NSCs-exs treatment in 3D culture. Exosome-guided differentiation on 3D PAN scaffolds promotes neural lineage commitment, reinforcing exosomes as reparative tools for neural regeneration.