Chitin-based tubes for tissue engineering in the nervous system

The purpose of this study was to investigate chitin and chitosan as potential materials for biodegradable nerve guides. Transparent chitin hydrogel tubes were synthesized, for the first time, from chitosan solutions using acylation chemistry and mold casting techniques. Alkaline hydrolysis of chitin tubes resulted in chitosan tubes, with the extent of hydrolysis controlling the resulting amine content. This, in turn, impacted compressive strength and cell adhesion. Chitosan tubes were mechanically stronger than their chitin origins, as measured by the transverse compressive test, where tubes having degrees of acetylation of 1%, 3%, 18% (i.e. chitosan) and 94% (i.e. chitin) supported loads at a 30% displacement of 40.6±4.3, 25.3±4.5, 10.6±0.8, and 8.7±0.4 g, respectively. However, the chitin processing methodology could be optimized for compressive strength, by either incorporating reinforcing coils in the tube wall, or air-drying the hydrogel tubes. Chitin and chitosan supported adhesion and differentiation of primary chick dorsal root ganglion neurons in vitro. Chitosan films showed significantly enhanced neurite outgrowth relative to chitin films, reflecting the dependence of nerve cell affinity on the amine content in the polysaccharide: neurites extended 1794.7±392.0 μm/mm2 on chitosan films vs. 140.5±41.6 μm/mm2 on chitin films after 2 days of culture. This implies that cell adhesion and neurite extension can be adjusted by amine content, which is important for tissue engineering in the nervous system. The methods for easy processing and modification of chitin and chitosan described herein, allow the mechanical properties and cyto-compatibility to be controlled and provide a means for a broader investigation into their use in biomedical applications.