Functionalization and peptide-based delivery of magnetic nanoparticles as an intracellular MRI contrast agent

We report the development of functionalized superparamagnetic iron oxide nanoparticles with a PEG-modified, phospholipid micelle coating, and their delivery into living cells. The size of the coated particles, as determined by dynamic light scattering and electron microscopy, was found to be between 12 and 14 nm. The PEG-phospholipid coating resulted in high water solubility and stability, and the functional groups of modified PEG allowed for bioconjugation of various moieties, including a fluorescent dye and the Tat peptide. Efficient delivery of the functionalized nanoparticles into living cells was confirmed by fluorescence microscopy, relaxation time measurements, and magnetic resonance imaging (MRI). This demonstrates the feasibility of using functionalized magnetic nanoparticles with uniform (~10 nm) sizes as an MRI contrast agent for intracellular molecular imaging in deep tissue. These micelle-coated iron oxide nanoparticles offer a versatile platform for conjugation of a variety of moieties, and their small size confers advantages for intracellular molecular imaging with minimal perturbation.Abbreviations CPP cell penetrating peptide - CPMG Carr–Purcell–Meiboom–Gill spin-echo method - CTAB cetyltrimethylammonium bromide - DLS dynamic light scattering - DMEM Dulbeccos modified Eagles medium - DSPE 1,2-distearoyl-sn-glycero-3-phosphoethanolamine - FCS fetal calf serum - FGM-2 fibroblast growth medium 2 - HDF human dermal fibroblast - HS horse serum - MDBK Madin–Darby bovine kidney - MIONs superparamagnetic iron oxide nanoparticles - mMIONs micelle-coated MIONs - MRI magnetic resonance imaging - PBS phosphate-buffered saline - PEG poly(ethylene glycol) - SPDP N-succinimidyl 3-(2-pyridyldithio)propionate - TCEP tris(2-carboxyethyl)phosphine hydrochloride - TEM transmission electron microscopy