Encapsulating a Single Nanoprobe in a Multifunctional Nanogel for High-Fidelity Imaging of Caspase Activity in Vivo.

The formation of a protein corona on nanoprobes in the blood can not only reduce the delivery efficiency to their destination but also inhibit the functions of the nanoprobes. Herein, we report a multifunctional nanogel that can shield a single gold nanoparticle (AuNP) probe from interaction with the serum proteins, virtually eliminating protein corona formation on the nanoprobes. As a result, the delivery efficiency of the nanogel-encapsulated nanoprobes to tumors was dramatically enhanced. When the probes are delivered into target cells, the nanogel shells are degraded in acidic endosomes, where a proton sponge effect occurs instantaneously to release the AuNP probes into the cytoplasm to realize their bioimaging functions. We demonstrated the applicability of these probes for high-fidelity, noninvasive imaging of caspase activity in both cancer cells and in tumors. This strategy offers an exciting opportunity to design high-efficacy nanoprobes for in vivo imaging.

Self-Assembly of Biocompatible FeSe Hollow Nanostructures and 2D CuFeSe Nanosheets with One- and Two-Photon Luminescence Properties.

Transition metal chalcogenides are investigated for catalyst, intermediary agency, and particular optical properties because of their distinguished electron-vacancy-transfer (EVT) process toward different applications. In this work, one convenient approach for making pure-phased FeSe nanocrystals (NCs) and doped CuFeSe nanosheets (NSs) through a wet chemistry method in mixed solvents is illustrated. The surface modification of each product is realized by using a peptide molecule glutathione (GSH), in which the thiol group (-SH) is ascribed to be the in situ reducer and bonding agency between the crystalline surface and surfactant in whole constructing processes. Due to the functional groups in biological GSH, highly aggregated NCs are rebuilt in the form of an FeSe hollow structure through amino and carboxyl cross-linking functions through a spontaneous assembly procedure. Owing to the coupling procedure of Cu and Fe in the growth process, it generates enhanced EVT. Additionally, it shows the emission spectra of λEM-PL = 436 nm (FeSe) and 452 nm (CuFeSe) while λEX-PL = 356 nm, it also conveys two-photon phenomenon while λEX-PL = 720 nm. Moreover, it also shows strong off-resonant luminescence due to two-photon absorption, which should be valuable for biological applications.

Reversibly extracellular pH controlled cellular uptake and photothermal therapy by PEGylated mixed-charge gold nanostars.

Shielding nanoparticles from nonspecific interactions with normal cells/tissues before they reach and after they leave tumors is crucial for the selective delivery of NPs into tumor cells. By utilizing the reversible protonation of weak electrolytic groups to pH changes, long-chain amine/carboxyl-terminated polyethylene glycol (PEG) decorated gold nanostars (GNSs) are designed, exhibiting reversible, significant, and sensitive response in cell affinity and therapeutic efficacy to the extracellular pH (pHe) gradient between normal tissues and tumors. This smart nanosystem shows good dispersity and unimpaired photothermal efficacy in complex bioenvironment at pH 6.4 and 7.4 even when their surface charge is neutral. One PEGylated mixed-charge GNSs with certain surface composition, GNS-N/C 4, exhibits high cell affinity and therapeutic efficacy at pH 6.4, and low affinity and almost "zero" damage to cells at pH 7.4. Remarkably, this significant and sensitive response in cell affinity and therapeutic efficacy is reversible as local pH alternated. In vivo, GNS-N/C 4 shows higher accumulation in tumors and improved photothermal therapeutic efficacy than pH-insensitive GNSs. This newly developed smart nanosystem, whose cell affinity reversibly transforms in response to pHe gradient with unimpaired biostability, provides a novel effective means of tumor-selective therapy.

Engineered Cancer-Derived Small Extracellular Vesicle-Liposome Hybrid Delivery System for Targeted Treatment of Breast Cancer.

Cancer-derived small extracellular vesicles (sEVs) may be a promising drug delivery system that targets cancer cells due to their unique features, such as native homing ability, biological barrier crossing capability, and low immune response. However, the oncogenic cargos within them pose safety concerns, hence limiting their application thus far. We proposed using an electroporation-based strategy to extract the endogenous cargos from cancer-derived sEVs and demonstrated that their homing ability was still retained. A membrane fusion technique was used to fuse these sEVs with liposomes to form hybrid particles, which possessed both benefits of sEVs and liposomes. Anti-EGFR monoclonal antibodies were modified on the hybrid particles to improve their targeting ability further. The engineered hybrid particles showed higher drug loading ability that is 33.75 and 43.88% higher than that of liposomes and sEVs, respectively, and improved targeting ability by 52.23% higher than hybrid particles without modification. This delivery system showed >90% cell viability and enhanced treatment efficiency with 91.58 and 79.26% cell migration inhibition rates for the miR-21 inhibitor and gemcitabine, respectively.

Using self-polymerized dopamine to modify the antifouling property of oligo(ethylene glycol) self-assembled monolayers and its application in cell patterning.

We report a one-step, mild method to modify antifouling oligo(ethylene glycol)-terminated self-assembled monolayers. We demonstrate for the first time that self-polymerized dopamine, previously reported as an underwater adhesive, can be patterned on typical antifouling surfaces by microfluidic patterning or microcontact printing. The patterns can be applied in spatiotemporal cell patterning.

Release Strategies of Silver Ions from Materials for Bacterial Killing.

Silver-based antibacterial agents have attracted much attention in biomedical fields owing to their high antibacterial activity. Silver ions (Ag+) play a significant role in killing bacteria because they can readily adsorb to most biomolecules (DNA, membrane protein, enzymes, or intracellular cofactors) in bacteria to inactivate their functions. In this review, recent advances in the ways of Ag+ release from silver-contained materials were discussed. As a start, the antibacterial mechanisms of Ag+ was summarized. Then, the release strategies of Ag+ from silver-carrying materials have been categorized into two types: (1) passive release and (2) stimuli-responsive release. In particular, the stimuli-responsive materials relying on either endogenous or exogenous stimuli have been rationally designed for eliminating bacteria with high efficacy and selectivity. Future studies should focus on enhancing the controllability of Ag+ release in the infected sites of the human organs.

In vitro model on glass surfaces for complex interactions between different types of cells.

This report establishes an in vitro model on glass surfaces for patterning multiple types of cells to simulate cell-cell interactions in vivo. The model employs a microfluidic system and poly(ethylene glycol)-terminated oxysilane (PEG-oxysilane) to modify glass surfaces in order to resist cell adhesion. The system allows the selective confinement of different types of cells to realize complete confinement, partial confinement, and no confinement of three types of cells on glass surfaces. The model was applied to study intercellular interactions among human umbilical vein endothelial cells (HUVEC), PLA 801 C and PLA801 D cells.

Reversible Self-Assembly of Nanoprobes in Live Cells for Dynamic Intracellular pH Imaging.

Self-assembly is a powerful tool to organize the elementary molecular units into functional nanostructures, which provide reversible stimulus-responsive systems for a variety of purposes. However, the ability to modulate the reversible self-assembly in live systems remains a great challenge owing to the chemical complexity of intracellular environments, which often damage synthetic assembled superstructures. Herein, we describe a robust reversible self-assembly system that is composed of a hydrophobic gold nanoparticle (AuNP) core and a shell of pH-responsive dye-incorporated block copolymers. The reversible assembly-disassembly processes were precisely controlled through mediating the molecular interactions between the copolymers and AuNPs. More importantly, the major endogenous biospecies such as proteins will not impair the reversible self-assembly, which was supported by free-energy calculations. The reversible pH-responsive nanostructures were employed as "smart" probes for visualizing the subtle dynamic pH changes among different intracellular compartments, facilitating the study of pH influence on biological processes.

An Ultrasensitive Biosensing Platform Employing Acetylcholinesterase and Gold Nanoparticles.

Enzyme-linked immunosorbent assay (ELISA) is a well-known strategy for biomarker detection with a color change, which can be seen by the naked eyes. However, the moderate sensitivity of conventional ELISA limits its applications in many cases where the concentrations of biomarker are very low, such as cancer diagnosis. Here we describe an ultrasensitive colorimetric assay based on acetylcholinesterase (AChE)-catalyzed hydrolysis reaction, whose products trigger the aggregation of gold nanoparticles (AuNPs), causing a distinct color change of the solution from red to purple. This enhanced colorimetric immunoassay offers extremely high sensitivity and specificity. In this study, we employed enterovirus 71 (EV71), the major cause of hand, foot, and mouth disease (HFMD), as a model to evaluate the analytical performance of the plasmonic immunoassay.

Tunable rigidity of (polymeric core)-(lipid shell) nanoparticles for regulated cellular uptake.

Core-shell nanoparticles (NPs) with lipid shells and varying water content and rigidity but with the same chemical composition, size, and surface properties are assembled using a microfluidic platform. Rigidity can dramatically alter the cellular uptake efficiency, with more-rigid NPs able to pass more easily through cell membranes. The mechanism accounting for this rigidity-dependent cellular uptake is revealed through atomistic-level simulations.

Chelator-free (64)Cu-integrated gold nanomaterials for positron emission tomography imaging guided photothermal cancer therapy.

Using positron emission tomography (PET) imaging to monitor and quantitatively analyze the delivery and localization of Au nanomaterials (NMs), a widely used photothermal agent, is essential to optimize therapeutic protocols to achieve individualized medicine and avoid side effects. Coupling radiometals to Au NMs via a chelator faces the challenges of possible detachment of the radiometals as well as surface property changes of the NMs. In this study, we reported a simple and general chelator-free (64)Cu radiolabeling method by chemically reducing (64)Cu on the surface of polyethylene glycol (PEG)-stabilized Au NMs regardless of their shape and size. Our (64)Cu-integrated NMs are proved to be radiochemically stable and can provide an accurate and sensitive localization of NMs through noninvasive PET imaging. We further integrated (64)Cu onto arginine-glycine-aspartic acid (RGD) peptide modified Au nanorods (NRs) for tumor theranostic application. These NRs showed high tumor targeting ability in a U87MG glioblastoma xenograft model and were successfully used for PET image-guided photothermal therapy.

A microfluidic origami chip for synthesis of functionalized polymeric nanoparticles.

This report demonstrates a microfluidic origami chip to synthesize monodisperse, doxorubicin-loaded poly(lactic-co-glycolic acid) nanoparticles with diameters of ~100 nm, a size optimized for cellular uptake and anticancer efficacy, but difficult to achieve with existing approaches. This three-dimensional design in a microchannel may allow for the fabrication of polymeric nanoparticles in this size regime with ease.

Dual-factor triggered fluorogenic nanoprobe for ultrahigh contrast and subdiffraction fluorescence imaging.

Ultrahigh contrast fluorescence molecular imaging has long been pursued over the past few decades from basic sciences to clinics. Although new classes of near-infrared (NIR) molecular probes are emerging, the requirement of fluorophores with high quantum yield, high signal to noise (S/N) ratio, and being activatable to microenvironment changes can hardly be fulfilled. In this study, a new NIR dye embedded fluorogenic nanoprobe (fg-nanoprobe) was developed for ultrahigh contrast in vitro and in vivo imaging with negligible background interference. The achieved S/N ratio was found to be attributed to the synergistic effects of the cellular compartmental triggered fluorogenicity and pH tunable fluorescence on/off character. In addition, this constructed fluorogenic nanoprobe could be coupled with image processing method for super-resolution subdiffraction imaging. The developed fg-nanoprobe integrated photophysical merits of the synthesized NIR fluorophore and advantages of engineered nanoparticle for enhanced fluorescence molecular imaging. This probe may open another avenue for ultrahigh contrast fluorescence molecular imaging in the future.

Highly robust, recyclable displacement assay for mercuric ions in aqueous solutions and living cells.

We designed a recyclable Hg(2+) probe based on Rhodamine B isothiocyanate (RBITC)-poly(ethylene glycol) (PEG)-comodified gold nanoparticles (AuNPs) with excellent robustness, selectivity, and sensitivity. On the basis of a rational design, only Hg(2+) can displace RBITC from the AuNP surfaces, resulting in a remarkable enhancement of RBITC fluorescence initially quenched by AuNPs. To maintain stability and monodispersity of AuNPs in real samples, thiol-terminated PEG was employed to bind with the remaining active sites of AuNPs. Besides, this displacement assay can be regenerated by resupplying free RBITC into the AuNPs solutions that were already used for detecting Hg(2+). Importantly, the detection limit of this assay for Hg(2+) (2.3 nM) was lower than the maximum limits guided by the United States Environmental Protection Agency as well as that permitted by the World Health Organization. The efficiency of this probe was demonstrated in monitoring Hg(2+) in complex samples such as river water and living cells.