title: Spinal cord organotypic slice cultures for the study of regenerating motor axon interactions with 3D scaffolds.
abstract: Numerous in-vitro techniques exist for investigating the influence of 3D substrate topography on sensory axon growth. However, simple and cost-effective methods for studying post-natal motor axon interactions with such substrates are lacking. Here, spinal cord organotypic slice cultures (OSC) from post-natal day 7-9 rat pups were presented with spinal nerve roots, or blocks of fibrin hydrogel or 3D microporous collagen scaffolds to investigate motor axon-substrate interactions. By 7-14 days, axons from motor neuronal pools extended into the explanted nerve roots, growing along Schwann cell processes and demonstrating a full range of axon-Schwann cell interactions, from simple ensheathment to concentric wrapping by Schwann cell processes and the formation of compact myelin within a basal lamina sheath. Extensive motor axon regeneration and all stages of axon-Schwann interactions were also supported within the longitudinally orientated microporous framework of the 3D collagen scaffold. In stark contrast, the simple fibrin hydrogel only supported axon growth and cell migration over its surface. The relative ease of demonstrating such motor axon regeneration through the microporous 3D framework by immunofluorescence, two-photon microscopy and transmission electron microscopy strongly supports the adoption of this technique for assaying the influence of substrate topography and functionalization in regenerative bioengineering.
authors.affiliation: Institute of Neuropathology, Uniklinik RWTH Aachen, Aachen 52074, Germany; Department of Translational Neuroscience, Maastricht University Medical Center, Maastricht 6229 ER, The Netherlands; European Graduate School of Neuroscience (EURON), Maastricht 6229 ER, The Netherlands. Electronic address: jgerardonava@ukaachen.de.
authors.forename: Jose
authors.initials: J
authors.lastname: Gerardo-Nava
authors.affiliation: Institute of Neuropathology, Uniklinik RWTH Aachen, Aachen 52074, Germany.
authors.forename: Dorothee
authors.initials: D
authors.lastname: Hodde
authors.affiliation: Institute of Neuropathology, Uniklinik RWTH Aachen, Aachen 52074, Germany.
authors.forename: Istvan
authors.initials: I
authors.lastname: Katona
authors.affiliation: Department of Plastic, Hand and Burns Surgery, Uniklinik RWTH Aachen University, Aachen 52074, Germany.
authors.forename: Ahmet
authors.initials: A
authors.lastname: Bozkurt
authors.affiliation: Department of Neurology, Ruhr-University Bochum, BG-Kliniken Bergmannsheil GmbH, Bochum 44789, Germany.
authors.forename: Torsten
authors.initials: T
authors.lastname: Grehl
authors.affiliation: Department of Translational Neuroscience, Maastricht University Medical Center, Maastricht 6229 ER, The Netherlands; European Graduate School of Neuroscience (EURON), Maastricht 6229 ER, The Netherlands.
authors.forename: Harry W M
authors.initials: HW
authors.lastname: Steinbusch
authors.affiliation: Institute of Neuropathology, Uniklinik RWTH Aachen, Aachen 52074, Germany; European Graduate School of Neuroscience (EURON), Maastricht 6229 ER, The Netherlands.
authors.forename: Joachim
authors.initials: J
authors.lastname: Weis
authors.affiliation: Institute of Neuropathology, Uniklinik RWTH Aachen, Aachen 52074, Germany; European Graduate School of Neuroscience (EURON), Maastricht 6229 ER, The Netherlands.
authors.forename: Gary A
authors.initials: GA
authors.lastname: Brook
mesh_terms: D000818:Animals; D001369:Axons / Q000473:pathology* / Q000648:ultrastructure; D018920:Coculture Techniques; D003094:Collagen / Q000378:metabolism; D005337:Fibrin / Q000494:pharmacology; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000494:pharmacology; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D009046:Motor Neurons / Q000187:drug effects / Q000473:pathology*; D009410:Nerve Degeneration / Q000473:pathology / Q000503:physiopathology; D009416:Nerve Regeneration / Q000187:drug effects; D009924:Organ Culture Techniques; D051381:Rats; D013116:Spinal Cord / Q000187:drug effects / Q000473:pathology / Q000503:physiopathology* / Q000648:ultrastructure; D013126:Spinal Nerve Roots / Q000187:drug effects / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D005337:Fibrin; D003094:Collagen
keywords: Collagen; Fibrin; In vitro test; Nerve regeneration; Scaffold