Temperature-sensitive micrometer-thick layers of hyaluronan grafted on microspheres

The giant polyelectrolyte glycosaminoglycan hyaluronan (1-10 MDa) is a major component of the pericellular coat on a variety of cells, where it is an important modulator and mediator of early cell adhesion events. This pericellular layer can reach 5 mu m thickness on cells that produce cartilage (chondrocytes), and up to 2 mu m on Xenopus laevis kidney epithelial cells (M). We are interested in generating model systems for the pericellular coat in order to learn more about the structure and function of hyaluronan on biological or artificial surfaces. We report here the synthesis of model systems where a coat of coordinatively cross-linked hyaluronan of up to 2 mu m thickness was covalently photografted onto polystyrene microspheres. The hydrated coat was imaged directly by environmental scanning electron microscopy (ESEM) at close to 100% relative humidity. The key feature of the procedure is the reversible reverse-temperature phase transition of hyaluronan induced by trivalent lanthanide cations, which is exploited to achieve sufficient density for grafting of thick layers. The microsphere-grafted coat shows a temperature-dependent swelling when labeled with lanthanide ions (Gd3+ or Tb3+). We directly observed a volume contraction of 20% with increasing temperature between 1 and 11 degrees C by wet-mode ESEM.