Controlled formation of cross-linked collagen fibers for neural tissue engineering applications

Fibrous scaffolds engineered to direct the growth of tissues can be important in forming architecturally functional tissue such as aligning regenerating nerves with their target. Collagen is a commonly used substrate used for neuronal growth applications in the form of surface coatings and hydrogels. The wet spinning technique can create collagen fibers without the use of organic solvents and is typically accomplished by extruding a collagen dispersion into a coagulation bath. To create well-controlled and uniform collagen fibers, we developed an automatic wet spinning device with precise control over the spinning and fiber collection parameters. A fiber collection belt allowed the continuous formation of very soft and delicate fibers up to half a meter in length. Wet-spun collagen fibers were characterized by tensile and thermal behavior, diameter uniformity, the swelling response in phosphate buffered saline and their biocompatibility with dorsal root ganglion (DRG) neurons and Schwann cells. Fibers formed from 0.75% weight by volume (w/v) collagen dispersions formed the best fibers in terms of tensile behavior and fiber uniformity. Fibers post-treated with the cross-linkers glutaraldehyde and genipin exhibited increased mechanical stability and reduced swelling. Importantly, genipin-treated fibers were conducive to DRG neurons and Schwann cell survival and growth, which validated the use of this cross-linker for neural tissue engineering applications.