Fluorescence imaging in genipin crosslinked chitosan-poly(vinyl pyrrolidone) hydrogels

Abstract

Recent research has identified genipin as a promising natural crosslinking agent for biocompatible hydrogels as genipin is significantly less cytotoxic than current synthetic crosslinking agents, such as glutaraldehyde. Conveniently, fluorophores can be produced when genipin crosslinks. In this study, fluorescence intensity measurements of genipin crosslinked chitosan-poly(vinyl pyrrolidone) hydrogels have been explored as a dynamic, in situ method for tracing sol-gel transition. These pH-responsive smart materials have a future in medical applications, in particular in tissue engineering and drug delivery, where methods to follow the process in situ and in real-time are crucial for future advancement. Samples were prepared using deionised water, pH 4, and pH 10 solutions, and studied at 24 and 37 °C over a 24 h period. Both temperature and pH have been found to affect sol-gel transition in the hydrogels studied. The transition from acidic (pH 4) to basic (pH 10) solution resulted in reduced fluorescence intensity suggesting that, under more basic conditions, genipin molecules self-polymerise, reducing the number of molecules available for reaction with the amino groups of chitosan. Three-dimensional representations of the fluorescence present in a hydrogel sample have also been produced from the data, enabling the visualisation of variation in fluorescence with time at the surface of the hydrogel.