Biodegradable AuNP-Based Plasmonic Nanogels as Contrast Agents for Computed Tomography and Photoacoustics

Gold nanoparticles (AuNP) are well-established contrast agents in computed tomography (CT) and photoacoustic imaging (PAI). A wide variety of AuNP sizes, shapes, and coatings have been reported for these applications. However, for clinical translation, AuNP should be excretable to avoid long-term accumulation and possible side effects. Sub-5 nm AuNP have the benefit to be excretable through kidney filtration, therefore their loading in biodegradable nanogels holds promise to result in contrast agents that have long circulation times in the vasculature and subsequent biodegradation for excretion. Polyphosphazenes are intrinsically biodegradable polymers capable of forming nanogels with high payloads, and to release their payloads upon degradation. The significant development in polyphosphazenes that have tailored degradation kinetics, and their formulation with drugs or contrast agents, has shown potential as a biodegradable platform for imaging vasculature and endogenous molecules, by combination of CT and PA modalities. Therefore, we herein present methods for the formulation of AuNP assemblies loaded in nanogels composed of biodegradable polyphosphazenes, with a size range from 50 to 200 nm. We describe protocols for their characterization by UV-vis spectroscopy, Fourier-transform infrared spectroscopy, various microscopy techniques, elemental quantification by induced coupling plasma optical emission spectroscopy and contrast production in both CT and PAI. Finally, we detail the methods to investigate their effect on cells, distribution in cells and imaging properties for detection of endogenous molecules.