Silencing Red Blood Cell Recognition toward Anti-A Antibody by Means of Polyelectrolyte Layer-by-Layer Assembly in a Two-Dimensional Model System

Abstract

Silencing the antigenic response of red blood cells (RBCs) is a prerequisite toward the development of universal blood transfusion. Using a two-dimensional (2D) model whereby nonfixed RBCs are adsorbed on a human fibronectin (HFN)-coated surface, we demonstrate that the layer-by-layer (LbL) assembly technique of biocompatible polyelectrolytes can be employed to achieve the immunocamouflage of RBCs against the Anti-A antibody while maintaining the integrity and viability of the cells. The multilayered film consisted of a protecting shell (P-shell), containing five bilayers of chitosan-graft-phosphorylcholine (CH-PC) and sodium hyaluronate (HA), covered by a camouflage shell (C-shell) made up of five bilayers of poly-(L-lysine)-graft-poly(ethylene glycol) (PLL-PEG) and alginate (AL). Control experiments in which RBCs were coated by (CH-PC/HA)(10) bilayers indicated that the two polyelectrolytes alone did not prevent immunorecognition. The LbL film formation on RBCs and model substrates was monitored by quartz crystal microbalance with dissipation factor (QCM-D) and analyzed through zeta-potential measurements, atomic force microscopy (AFM), and optical microscopy. Antibody interaction with the coated RBCs was investigated by QCM-D, fluorescence microscopy, and hemolysis assays. Results from these measurements demonstrated that the hybrid LbL system built-up with different sets of polyelectrolytes was able to protect the RBCs from hemolysis and recognition by the Anti-A antibody.

Supported Lipid Bilayers on Biocompatible Polysaccharide Multilayers

“…22, 23 The biocompatibility, ready availability and low cost of charged polysaccharides have made them attractive choices for a range of applications, including drug delivery vehicles based on lipid-polyelectrolyte particles, biocompatible surfaces for controlling cell growth and tissue biomimetics. [23][24][25][26][27][28] We have examined the suitability of PEM films constructed from alternating layers of chitosan (CHI) and hyaluronic acid (HA) as polymer cushions for supported lipid bilayers.…”

Enhanced MC3T3 preosteoblast viability and adhesion on polyelectrolyte multilayer films composed of glycol‐modified chitosan and hyaluronic acid

“…Recent work with cast films composed of water‐soluble phosphorylcholine‐modified chitosan (Pc‐CHI) has indicated significantly improved bone marrow cellular adhesion properties in comparison to unmodified CHI 19. Meanwhile, initial studies of LbL films composed of Pc‐CHI and HA have demonstrated biocompatibility 20, 21. The more commercially available glycol‐modified chitosan (Glyc‐CHI) also exhibits significantly improved water solubility and biocompatibility, features that have led to its wide deployment in drug delivery systems, as well as in gene therapy and tissue engineering applications 22–25.…”

Molecular Engineering of Cell and Tissue Surfaces with Polymer Thin Films

“…Tabrizian and colleagues [64,97] have recently taken a different polymer film engineering approach to generating a universal RBC. Instead relying on a single polymer layer to form a steric barrier between RBC surface antigens and antibodies, they instead assembled a PEM thin film on the surface of RBCs.…”