Homogeneous films from amphiphilic hyaluronan and their characterization by confocal microscopy and nanoindentation

Self-supporting films from amphiphilic hyaluronan are suitable for medical applications like wound dressings or resorbable implants. These films are typically cast from water/alcohol solutions. However, when the mixed solvent evaporates in ambient air, convection flows develop in the solution and become imprinted in the film, potentially compromising its properties. Consequently, we developed a novel film manufacturing method: drying in a closed box under saturated vapour conditions. Using this approach, we prepared a series of optically clear lauroyl-hyaluronan (LHA) films with uniform thickness and compared them to their air-dried counterparts. We first evaluated swelling ratios and elastic moduli for LHA films with varying degrees of substitution. The box-dried films swelled significantly less and were 1-2 orders of magnitude stiffer than air-dried films from the same LHA sample. Confocal microscopy revealed that box-dried films exhibited a regular microstructure, while air-dried films displayed a pore-size gradient and strong microstructure modulation due to convection flows. Local elastic modulus variations arising from these microstructures were assessed using nanoindentation mapping. Importantly, achieving the desired film stiffness requires much lower polymer modification when box-drying is used, enhancing the biological response to the material. These findings have implications for all polysaccharide formulations that utilize mixed solvents.