No abstract

Immune cells take up nanoscale materials and can be programmed to release it again, which has important implications for understanding cellular functions, biocompatibility as well as biomedical applications.

Transport and delivery of (nanoscale) materials are crucial for many applications in biomedicine. However, controlled uptake, transport and triggered release of such cargo remains challenging. In this study, we use human immune cells (neutrophilic granulocytes, neutrophils) and program them to perform these tasks in vitro. For this purpose, we let neutrophils phagocytose a nanoscale cargo. As an example, we used DNA-functionalized single-walled carbon nanotubes (SWCNT) that fluoresce in the near infrared (980 nm) and serve as sensors for small molecules. Cells still migrate, follow chemical gradients and respond to inflammatory signals after uptake of the cargo. To program release, we make use of neutrophil extracellular trap formation (NETosis), a novel cell death mechanism that leads to chromatin swelling and subsequent rupture of the cellular membrane and release of the cell's whole content. By using the process of NETosis we can program the time point of cargo release via the initial concentration of stimuli such as phorbol 12-myristate-13-acetate (PMA) or lipopolysaccharide (LPS). At intermediate stimulation with LPS (100 µg/ml), cells continue to migrate, follow gradients and surface cues for around 30 minutes and up to several hundred micrometers until they stop and release their cargo. The transported and released SWCNT sensor cargo is still functional as shown by subsequent detection of the neurotransmitter dopamine and reactive oxygen species (H 2 O 2 ). In summary, we hijack a biological process (NETosis) and demonstrate how neutrophils can be used for programmed transport and delivery of functional nanomaterials. 3