Layer-by-Layer Surface Modification of Alendronate-Loaded Polyester Microparticles—Enabling Protein Immobilization

Abstract

The highly inert surface of polyester micro- and nano- drug carriers is a challenging substrate for further modification. The presence of surface moieties suitable for macromolecule coupling is crucial in the development of targeted drug delivery systems. Among available methods of surface activation, those based on adsorption of charged macromolecules may be carried out in mild conditions. In this work, alendronate-loaded microcores of three polyesters: poly-ε-caprolactone (PCL), poly(l-lactide-co-ε-caprolactone) (PLA-co-PCL) and poly(lactic-co-glycolic acid) (PLGA) were coated with three polyelectrolyte shells composed of chitosan/heparin (CHIT/HEP), polyallylamine/heparin (PAH/HEP), and polyethyleneimine/heparin (PEI/HEP) via the layer-by-layer method. Subsequently, the feasibility of model protein immobilization on obtained shells was assessed. Electrokinetic potential measurements confirmed the possibility of deposition of all investigated coating variants, and a positive correlation between initial core ζ potential and intensity of charge alterations after deposition of subsequent layers was identified. PEI/HEP assembly was stable in physiological-like conditions, while PAH/HEP multilayers disassembled in presence of phosphate ions, and CHIT/HEP shell showed limited stability in pH 7.4. Fluorescence assays of fluorescein tagged lysozyme surface coupled via ethylcarbodiimide hydrochloride/N-Hydroxysuccinimide (EDC/NHS) click reaction with all shell variants indicated satisfying reaction efficiency. Poly-ε-caprolactone cores coated with CHIT/HEP tetralayer were selected as suitable for model IgG surface immobilization. Antibodies immobilized on the shell surface exhibited a moderate degree of affinity to fluorescent IgG binding protein.