A polyelectrolyte multilayer platform for investigating growth factor delivery modes in human liver cultures

Abstract

Polyelectrolyte multilayers (PEMs) of chitosan and heparin are useful for mimicking growth factor (GF) binding to extracellular matrix (ECM) as in vivo. Here, we developed a PEM platform for delivering bound/adsorbed GFs to monocultures of primary human hepatocytes (PHHs) and PHH/non-parenchymal cell (NPC) co-cultures, which are useful for drug development and regenerative medicine. The effects of ECM protein coating (collagen I, fibronectin, and Matrigel®) and terminal PEM layer on PHH attachment/functions were determined. Then, heparin-terminated/fibronectin-coated PEMs were used to deliver varying concentrations of an adsorbed model GF, transforming growth factor β (TGFβ), to PHH monocultures while using soluble TGFβ delivery via culture medium as the conventional control. Soluble TGFβ delivery caused a severe, monotonic, and sustained downregulation of all PHH functions measured (albumin and urea secretions, cytochrome-P450 2A6 and 3A4 enzyme activities), whereas adsorbed TGFβ delivery caused transient upregulation of 3 out of 4 functions. Finally, functionally stable co-cultures of PHHs and 3T3-J2 murine embryonic fibroblasts were created on the heparin-terminated/fibronectin-coated PEMs modified with adsorbed TGFβ to elucidate similarities and differences in functional response relative to the monocultures. In conclusion, chitosan-heparin PEMs constitute a robust platform for investigating the effects of GF delivery modes on PHH monocultures and PHH/NPC co-cultures. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 971-984, 2018.

Progress in the Development of Chitosan-Based Biomaterials for Tissue Engineering and Regenerative Medicine

“…The polycationic nature of CS is used to prepare polyelectrolyte complexes which spontaneously form upon mixing. For example, heparin is widely used with CS polyelectrolyte complex due to its ability to bind growth factors and cytokines [113][114][115][116][117]. In a recent study, CS-heparin NPs were used for the delivery of siRNA against vascular endothelial growth factor in human retinal epithelial cells (ARPE-19) with a 2-fold higher transfection efficiency in comparison to carrying plasmid DNA alone [118].…”

Biocompatible Crosslinked Nanofibers of Poly(Vinyl Alcohol)/Carboxymethyl‐Kappa‐Carrageenan Produced by a Green Process

“…The ECM is composed of several types of proteins, polysaccharides and glycoproteins, such as hyaluronic acid, fibrous collagen, laminin, fibronectin, and proteoglycans. [1,2] These components are continuously synthesized, secreted, arranged, and tailored by the cells that they support. Considering the fundamental roles of ECM in cell and tissue processes, the structure and chemistry of the ECM serves as a model for fabricating tissue engineering scaffolds and materials for studying cell-ECM interactions.…”

Enhanced hemocompatibility and antibacterial activity on titania nanotubes with tanfloc/heparin polyelectrolyte multilayers

“…PEMs can be easily and reproducibly prepared, to achieve control over coating thickness and surface chemistry, without the use of hazardous organic solvents (Almodóvar, Place, Gogolski, Erickson, & Kipper, ). Among the polycation‐polyanion polymer pairs recently used, heparin and chitosan PEMs have generated particular interest for biological activation of surfaces (Almodóvar et al, ; Lin et al, ; Zomer Volpato et al, ). Heparin (HP) is a highly negatively charged polymer and a natural glycosaminoglycan (GAG), which in the body prevents blood coagulation by inhibiting the thrombin activation (Zhang et al, ).…”