In Situ Labeling of the Aqueous Compartment of Extracellular Vesicles with Luminescent Gold Nanoclusters.

Extracellular vesicles (EVs) are well-known membrane-limited particles secreted by both healthy and cancerous cells. They are considered as biomarkers for early cancer diagnosis and are involved in many pathologies and physiological pathways. They could serve as diagnostic tools in liquid biopsies, as therapeutics in regenerative medicine, or as drug delivery vehicles. Our aim is here to encapsulate luminescent nanoprobes in the aqueous compartment of human EVs extracted from reproductive fluids. The analysis and labeling of the EVs content with easily detectable luminescent nanoparticles could enable a powerful tool for early diagnosis of specific diseases and also for the design of new therapeutics. In this view, gold nanoclusters (AuNCs) appear as an attractive alternative as nontoxic fluorophore probes because of their luminescence properties, large window of fluorescence lifetimes (1 ns-1 µs), ultrasmall size (<2 nm), good biocompatibility, and specific ability as X-ray photosensitizers. Here, we investigated an attractive method that uses fusogenic liposomes to deliver gold nanoclusters into EVs. This approach guarantees the preservation of the EVs membrane without any breakage, thus maintaining compartmental integrity. Different lipid compositions of liposomes preloaded with AuNCs were selected to interact electrostatically with human EVs and compared in terms of fusion efficiency. The mixture of liposomes and EVs results in membrane mixing as demonstrated by FRET experiments and fusion revealed by flux cytometry and cryo-TEM. The resulting fused EVs exhibit typical fluorescence of the AuNCs together with an increased size in agreement with fusion. Moreover, the fusion events in mixtures of EVs and AuNCs preloaded liposomes were analyzed by using cryo-electron microscopy. Finally, the ratio of released AuNCs during the fusion between the fusogenic liposomes and the EVs was estimated to be less than 20 mol % by Au titration using ICP spectroscopy.

Internalization of Pegylated Er:Y2O3 Nanoparticles inside HCT-116 Cancer Cells: Implications for Imaging and Drug Delivery.

Lanthanide-doped nanoparticles, featuring sharp emission peaks with narrow bandwidth, exhibit high downconversion luminescence intensity, making them highly valuable in the fields of bioimaging and drug delivery. High-crystallinity Y2O3 nanoparticles (NPs) doped with Er3+ ions were functionalized by using a pegylation procedure to confer water solubility and biocompatibility. The NPs were thoroughly characterized using transmission electron microscopy (TEM), inductively coupled plasma mass spectrometry (ICP-MS), and photoluminescence measurements. The pegylated nanoparticles were studied both from a toxicological perspective and to demonstrate their internalization within HCT-116 cancer cells. Cell viability tests allowed for the identification of the "optimal" concentration, which yields a detectable fluorescence signal without being toxic to the cells. The internalization process was investigated using a combined approach involving confocal microscopy and ICP-MS. The obtained data clearly indicate the efficient internalization of NPs into the cells with emission intensity showing a strong correlation with the concentrations of nanoparticles delivered to the cells. Overall, this research contributes significantly to the fields of nanotechnology and biomedical research, with noteworthy implications for imaging and drug delivery applications.

Supramolecular Detection of a Sub-ppm Nerve Agent Simulant by a Smartphone Tool.

The widespread use of smartphones and related tools is extending their applications in several fields. Herein, we report a reusable smartphone coupled portable detection system for the sensing of sub-ppm level of a nerve agent mimic (dimethylmethylphosphonate) in the gas phase. The detection system is based on multiple hydrogen-bond interactions of the vapor analyte with an ad-hoc functionalized Bodipy chromophore scaffold. The multitopic approach used for the molecular recognition of DMMP leads to the highest binding constant values, high selectivity, and low limits of detection.

Supramolecular Sensing of Chemical Warfare Agents.

Chemical warfare agents are a class of organic molecules used as chemical weapons due to their high toxicity and lethal effects. For this reason, the fast detection of these compounds in the environment is crucial. Traditional detection methods are based on instrumental techniques, such as mass spectrometry or HPLC, however the use of molecular sensors able to change a detectable property (e. g., luminescence, color, electrical resistance) can be cheaper and faster. Today, molecular sensing of chemical warfare agents is mainly based on the "covalent approach", in which the sensor reacts with the analyte, or on the "supramolecular approach", which involves the formation of non-covalent interactions between the sensor and the analyte. This Review is focused on the recent developments of supramolecular sensors of organophosphorus chemical warfare agents (from 2013). In particular, supramolecular sensors are classified by function of the sensing mechanism: i) Lewis Acids, ii) hydrogen bonds, iii) macrocyclic hosts, iv) multi-topic sensors, v) nanosensors. It is shown how the supramolecular non-covalent approach leads to a reversible sensing and higher selectivity towards the selected analyte respect to other interfering molecules.