Journal
CARBOHYDRATE POLYMERS
Volume 91, Issue 1, Pages 410-418Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.carbpol.2012.08.066
Keywords
Hyaluronan; Hydrogel; Degree of cross-linking; Effective cross-linker ratio; Degree of modification; Characterization; Determination
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Definitions and methods for the quantification of degree of modification and cross-linking in cross-linked hyaluronic acid (HA) hydrogels are outlined. A novel method is presented in which the HA hydrogel is degraded by the enzyme chondroitinase AC and the digest product analyzed by size exclusion chromatography combined with electrospray ionization mass spectrometry (SEC-ESI-MS). This method allows for the determination of effective cross-linker ratio (CrR) which together with the degree of modification (MoD), determined by, e.g. H-1 NMR spectroscopy, enables the calculation of the degree of substitution (DS) and degree of cross-linking (CrD). The method, could be applicable to the major cross-linked HA hydrogels currently on the market, and is exemplified here by application to two HA hydrogels. The definitions and methods presented are important contributions in attempts to find relationships between MoD, DS and CrD to mechanical properties as well as to biocompatibility of HA hydrogels. (C) 2012 Elsevier Ltd. All rights reserved.
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