Enzymatic Degradation of Glycosaminoglycans and Proteoglycan-Mimetic Materials in Solution and on Polyelectrolyte Multilayer Surfaces.

Proteoglycans (PGs) play many important roles in biology, contributing to the mechanical properties of tissues, helping to organize extracellular matrix components, and participating in signaling mechanisms related to mechanotransduction, cell differentiation, immune responses, and wound healing. Our lab has designed two different types of PG mimics: polyelectrolyte complex nanoparticles (PCNs) and PG-mimetic graft copolymers (GCs), both of which are prepared using naturally occurring glycosaminoglycans. This work evaluates the enzymatic stability of these PG mimics using hyaluronidases (I-S, IV-S, and II), chondroitinase ABC, and lysozyme, for PG mimics suspended in solution and adsorbed onto surfaces. Hyaluronan (HA)- and chondroitin sulfate (CS)-containing PG mimics are degraded by the hyaluronidases. PCNs prepared with CS and GCs prepared with heparin are the only CS- and HA-containing PG mimics protected from chondroitinase ABC. None of the materials are measurably degraded by lysozyme. Adsorption to polyelectrolyte multilayer surfaces protects PG mimics from degradation, compared to when PG mimics are combined with enzymes in solution; all surfaces are still intact after 21 days of enzyme exposure. This work reveals how the stability of PG mimics is controlled by both the composition and macromolecular assembly of the PG mimic and also by the size and specificity of the enzyme. Understanding and tuning these degradation susceptibilities are essential for advancing their applications in cardiovascular materials, orthopedic materials, and growth factor delivery applications.

Polyelectrolyte multilayers containing a tannin derivative polyphenol improve blood compatibility through interactions with platelets and serum proteins.

To develop hemocompatible surfaces, a cationic tannin derivate (TN) was used to prepare polyelectrolyte multilayers (PEMs) with the glycosaminoglycans heparin (HEP) and chondroitin sulfate (CS). The surface chemistry of the PEMs was characterized using X-ray photoelectron spectroscopy and water contact angle measurements. PEMs assembled with chitosan (CHI) and HEP or CS were used as controls. We investigate the hemocompatibility of PEMs by analyzing the adsorption of key blood serum proteins, adhesion and activation of platelets, and blood clotting kinetics. TN- and CHI-based PEMs adsorb similar amounts of albumin, whereas fibrinogen adsorption was more pronounced on TN-based PEMs, due to strong association with catechol groups. However, TN-based PEMs significantly reduce both platelet adhesion and platelet activation, while CHI-based PEMs promote platelet adhesion and activation. The whole-blood clotting kinetics assay also shows lower blood coagulation on TN-based PEMs. TN is an amphoteric, cationic, condensed tannin derivative with resonance structures. It also contains catechol groups, which are similar to those in mussel adhesive protein. These chemical features enable strong association with fibrinogen, which promotes the platelet-repelling effect. This study provides a new perspective for understanding platelet adhesion and activation on biomaterial surfaces, toward the development of new blood-compatible surfaces using a tannin derivative-based polymer.