Polycationic Nanoparticles: (1) Synthesis of a Polylysine-MION Conjugate and its Application in Labeling Fibroblasts

Abstract

Nanoparticles are increasingly used to label cells to track them by imaging or to quantify them in vivo. However, normal cellular uptake mechanisms are inadequate to load cells with tracking label. We propose a simple method to coat nanoparticles, such as monocrystalline iron oxide nanoparticle (MION), with the transfection agent polylysine in order to facilitate rapid, uniform, and heavy labeling of fibroblasts. The method is based on commercially available reagents, requires no more than 1 h of laboratory contact time, and can be accomplished safely without a chemical hood. A suspension of MION was treated by addition of solid sodium periodate to oxidize glucose residues of dextran and introduced aldehyde groups to the dextran coat surrounding MION's crystalline magnetite core. After a 30-min incubation to effect oxidation, unreacted periodate was quenched with glycerol. The preparation was dialyzed to remove reactants and diluted to a final concentration of 2 mg Fe/ml. Poly-L-lysine was added to the oxidized MION (MION-A) to form reversible covalent Schiff base linkages. The resulting conjugate, a polylysine iron oxide nano-particle is abbreviated PLION. NIH3T3 fibroblasts labeled with either MION, MION-A, or MION plus polylysine showed minimal uptake of iron while cells labeled with PLION acquired a brown hue demonstrating strong labeling with iron. Microscopic assessment of iron labeling was confirmed using Prussian blue staining. In some cells, the concentration of iron was sufficiently high and localized to suggest association with cytoplasmic vacuoles. The nucleus of the cell was not labeled. Cell labeling increased when the ratio of polylysine to MION increased and with increasing amount of PLION.