title: In vitro assessment of axonal growth using dorsal root ganglia explants in a novel three-dimensional collagen matrix.
abstract: The goal of this study was the development of a bioartificial nerve guide to induce axonal regeneration in the peripheral nervous system (PNS). In this in vitro study, the ability of a novel, 3-dimensional (3D), highly oriented, cross-linked porcine collagen scaffold to promote directed axonal growth has been studied. Collagen nerve guides with longitudinal guidance channels were manufactured using a series of chemical and mechanical treatments with a patented unidirectional freezing process, followed by freeze-drying (pore sizes 20-50 microm). Hemisected rat dorsal root ganglia (DRG) were positioned such that neural and non-neural elements could migrate into the collagen scaffold. After 21 days, S100-positive Schwann cells (SCs) migrated into the scaffold and aligned within the guidance channels in a columnar fashion, resembling "Bands of Büngner." Neurofilament-positive axons (mean length +/- SD 756 microm +/- 318 microm, maximum 1496 microm) from DRG neurons entered the scaffold where the growth within the guidance channels was closely associated with the oriented SCs. This study confirmed the importance of SCs in the regeneration process (neurotrophic theory). The alignment of SCs within the guidance channels supported directional axonal growth (contact guidance theory). The microstructural properties of the scaffold (open, porous, longitudinal pore channels) and the in vitro data after DRG loading (axonal regeneration along migrated and columnar-aligned SCs resembling "Band of Büngner") suggest that this novel oriented 3D collagen scaffold serves as a basis for future experimental regeneration studies in the PNS.
authors.affiliation: Department of Plastic and Hand Surgery, Burn Center, University Hospital, RWTH Aachen University, Aachen, Germany. abozkurt@ukaachen.de
authors.forename: Ahmet
authors.initials: A
authors.lastname: Bozkurt
authors.forename: Gary A
authors.initials: GA
authors.lastname: Brook
authors.forename: Sven
authors.initials: S
authors.lastname: Moellers
authors.forename: Franz
authors.initials: F
authors.lastname: Lassner
authors.forename: Bernd
authors.initials: B
authors.lastname: Sellhaus
authors.forename: Joachim
authors.initials: J
authors.lastname: Weis
authors.forename: Michael
authors.initials: M
authors.lastname: Woeltje
authors.forename: Julian
authors.initials: J
authors.lastname: Tank
authors.forename: Christina
authors.initials: C
authors.lastname: Beckmann
authors.forename: Paul
authors.initials: P
authors.lastname: Fuchs
authors.forename: Leon Olde
authors.initials: LO
authors.lastname: Damink
authors.forename: Frank
authors.initials: F
authors.lastname: Schügner
authors.forename: Ingo
authors.initials: I
authors.lastname: Heschel
authors.forename: Norbert
authors.initials: N
authors.lastname: Pallua
mesh_terms: D000818:Animals; D001369:Axons / Q000502:physiology / Q000648:ultrastructure*; D018929:Cell Culture Techniques / Q000379:methods; D049108:Cell Enlargement; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism*; D005727:Ganglia, Spinal / Q000166:cytology* / Q000502:physiology / Q000637:transplantation*; D048091:Guided Tissue Regeneration / Q000295:instrumentation / Q000379:methods*; D009416:Nerve Regeneration / Q000502:physiology*; D051381:Rats; D011917:Rats, Inbred Lew; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D003094:Collagen