Structure, depolymerization, and cytocompatibility evaluation of glycol chitosan

Abstract

Glycol chitosan, a water soluble chitosan derivative being investigated as a new biomaterial, was fractionated via two different methods. Initial characterization of the glycol chitosan with (1)H NMR spectroscopy illustrated the presence of both secondary and tertiary amine groups, contradictory to its widely accepted structure. Fractionation of glycol chitosan with nitrous acid resulted in a significant reduction in the number average molecular weight, specifically, from 170 to approximately 7 kDa for a pH 3 and below. However, the reaction altered its chemical structure, as the secondary amine groups were converted to N-nitrosamines, which are potentially carcinogenic. An increase in the pH of the reaction limited this formation, but not entirely. Free radical degradation initiated with potassium persulfate was not as effective at reducing the molecular weight as the nitrous acid approach, yielding molecular weights around 12 kDa under the same molar ratio of degrading species, but did retain the structural integrity of the glycol chitosan. Additionally, control of the molecular weight appears feasible with potassium persulfate. When assessed in vitro for cytocompatibility, the polymer exhibited no toxicity on monolayer-cultured chondrocytes, and in fact stimulated cell growth at low concentrations.