Gold nanoparticles (AuNPs) hold great promise in nanomedicine, yet their successful clinical translation has not been realized. Some challenges include effective AuNP targeting and delivery to improve modulation of immune cells of interest while limiting potential adverse effects. In order to overcome these challenges, we must fully understand how AuNPs impact different immune cell subsets, particularly within the dendritic cell and T cell compartments. Herein, we show that polyethylene glycol coated (PEG) gold nanorods (AuNRs) and PEG AuNRs covered with a thin layer of silver (AuNR/Ag) may enhance the immune response towards immune suppression or activation. We also studied the ability to enhance CD4+ Foxp3+ Tregs in vitro using AuNRs functionalized with interleukin 2 (IL2), a cytokine that is important in Treg development and homeostasis. Our results indicate that AuNRs enhance different immune cells and that NP composition matters in immune targeting. This knowledge will help us understand how to better design AuNRs to target and enhance the immune system.
Plasmonic gap-enhanced Raman tags (GERTs) are new emerging nanoprobes that, based on their unique surface-enhanced Raman spectroscopy (SERS) signal, can play a major role in complex imaging and detection of biological systems. GERTs are generated from a metal core nanostructure and layered with one or more metal nanosized layers, encasing a Raman active molecule. The advantages of GERTs are enhanced surface plasmon and electromagnetic resonance, as well as inherent protection of the Raman active molecule from environmental deterioration that could reduce their spectroscopic signatures over time. In this study, we used in vitro three-dimensional (3D) spheroid cultures to demonstrate these advantages. 3D spheroids mimic the in vivo tumor microenvironment better than 2D culture, with abundant extracellular matrix and hypoxia inducing variability of pH and enzymatic reactions. Here, we report the use of GERTs in large pancreatic 3D spheroids (>500 μm in apparent diameter) for complex penetration visualization. Our combined imaging technique of enhanced darkfield microscopy and SERS was able to identify the presence and distribution of the GERTs within the 3D spheroid structure. The distribution of GERTs 2 hours after the nanorods’ incubation indicated accumulation, generally in the outermost layer of the spheroids but also, more randomly, in non-uniform patterns in deep layers of the 3D spheroids. These observations bring into question the mechanism of uptake and flow of the nanoparticles in function of their incubation time while demonstrating the promising potential of our approach. Additionally, the SERS signal was still detectable after 24 hours of incubation of GERTs with the 3D culture, indicating the stability of the Raman signal.
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