Preparation of chitin hydrogel under mild conditions

Both the amount of water and the number of calcium ions are main factors affecting the dissolution of chitin in calcium chloride dihydrate-saturated methanol (calcium solvent). The higher degree of N-acetylation of the chitin was also indicated by its higher solubility in calcium solvent. The chitin hydrogel was prepared by adding a large excess of water to the chitin solution with vigorous stirring, followed by extensive dialysis against water or by filtration to remove the methanol and calcium ions. The water content of the chitin hydrogel was approximately 94-96% (w/v) and could be controlled by centrifugation. The chitin gel was also prepared by the addition of a large excess of alcohol, such as ethanol and iso-propanol, and these protocols were found to be effective under anhydrous conditions because the alcohols were exchangeable with other organic solvents in solution. The chitin hydrogel was more susceptible to lysozyme than to chitinase, and showed and a poor susceptibility to chitosanase. A alpha-chitin-type crystalline structure was regenerated from chitin sheets prepared from both alpha-chitin and beta-chitin solutions in calcium solvent, but the beta-chitin-type sheet was formed from the beta-chitin hydrogel prepared by mechanical agitation in water. The alpha-chitin hydrogel solidified when thawed after freezing, but the beta-chitin hydrogel prepared by mechanical agitation maintained its gel form even after prolonged freezing. Animal studies revealed a low toxicity for the chitin sheet and an acceleration of epidermal cell regeneration.