Autoclaving Achieves pH-Neutralization, Hydrogelation, and Sterilization of Chitosan Hydrogels in One Step

Conventionally, chitosan hydrogels are acidic and contain toxic chemicals because chitosan is soluble only in acidic solvents and requires toxic additives such as chemical crosslinkers and polymerization agents to fabricate chitosan hydrogels. These properties prevent chitosan hydrogels from being used for medical applications. In this study, chitosan hydrogels were prepared by a simple and versatile process using urea hydrolysis by autoclaving (steam sterilization, 121 degrees C, 20 min). When autoclaved, urea hydrolyzes in an acidic chitosan aqueous solution, and ammonia is produced, which increases the pH of the solution, and chitosan becomes insoluble, leading to the formation of a chitosan hydrogel. The pH and osmotic concentration of chitosan hydrogels could be adjusted to be suitable for physiological conditions (pH: 7.0-7.5, and osmotic concentration: 276-329 mOsm/L) by changing the amount of urea added to chitosan solutions (chitosan: 2.5% (w/v), urea: 0.75-1.0% (w/v), pH: 5.5). The hydrogels had extremely low cytotoxicity without the washing process. In addition, not only pure chitosan hydrogels, but also chitosan derivative hydrogels were prepared using this method. The autoclaving technique for preparing low-toxic and wash-free sterilized chitosan hydrogels in a single step is practical for medical applications.