Abstract
Hyaluronic acid (HA)/chitosan (CHI) complex coacervates have recently gained interest due to the pH-dependent ionization and semiflexibility of the polymers as well as their applicability in tissue engineering. Here, we apply isothermal titration calorimetry (ITC) to understand the apparent thermodynamics of coacervation for HA/CHI as a function of the pH, ionic strength, and chain length. We couple these ITC experiments with the knowledge of the charge states of HA and CHI from potentiometric titration to understand the mechanistic aspects of complex formation. Our data demonstrate that the driving force for the complex coacervation of HA and CHI is entropic in nature and this driving force decreased with increasing ionic strength. We also observed a decrease in the stoichiometry for ion-pairing with increasing ionic strength, which we suggest is a consequence of the changing degree of ionization for HA at higher ionic strengths. An increase in the strength of interactions with pH was hypothesized to also be a result of changes in the degree of ionization of HA, though stronger interactions were observed at the lowest pH tested, likely due to contributions from hydrogen bonding between HA and CHI.
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