Transplantable living scaffolds comprised of micro-tissue engineered aligned astrocyte networks to facilitate central nervous system regeneration

摘要: Abstract Neurotrauma, stroke, and neurodegenerative disease may result in widespread loss of neural cells as well the complex interconnectivity necessary for proper central nervous system function, generally resulting permanent functional deficits. Potential regenerative strategies involve recruitment endogenous stem and/or directed axonal regeneration through use tissue engineered “living scaffolds” built to mimic features three-dimensional (3-D) vivo migratory or guidance pathways. Accordingly, we devised a novel biomaterial encasement scheme using tubular hydrogel-collagen micro-columns that facilitated self-assembly seeded astrocytes into 3-D living scaffolds consisting long, cable-like aligned astrocytic networks. Here, robust astrocyte alignment was achieved within micro-column inner diameter (ID) 180 μm 300–350 μm but not 1.0 mm, suggesting radius curvature dictated extent alignment. Moreover, small ID micro-columns, >70% assumed bi-polar morphology, versus ∼10% larger planar surfaces. Cell–cell interactions also influenced architecture, extensive astrocyte-collagen contraction at high (9–12 × 105 cells/mL) lower (2–6 × 105 cells/mL) seeding densities. This density led formation ultra-dense “bundles” collagen measuring up 150 μm yet extending remarkable length over 2.5 cm. Importantly, co-seeded neurons extended neurites directly along bundles, demonstrating permissive cues neurite extension. These transplantable networks structurally glial tube guides neuronal progenitor migration rostral stream, therefore be useful guide repopulate sites neurodegeneration. Statement Significance manuscript details our development micro-tissue engineering techniques generate longitudinally hydrogels. We report miniature scaffold” constructs physically emulate – pathway brain facilitate repair. The size self-contained design these will permit minimally invasive transplantation models injury future studies.