Alkyl‐Capped Silicon Nanocrystals Lack Cytotoxicity and have Enhanced Intracellular Accumulation in Malignant Cells via Cholesterol‐Dependent Endocytosis

Nanocrystals of various inorganic materials are being considered for application in the life sciences as fluorescent labels and for such therapeutic applications as drug delivery or targeted cell destruction. The potential applications of the nanoparticles are critically compromised due to the well‐documented toxicity and lack of understanding about the mechanisms involved in the intracellular internalization. Here intracellular internalization and toxicity of alkyl‐capped silicon nanocrystals in human neoplastic and normal primary cells is reported. The capped nanocrystals lack cytotoxicity, and there is a marked difference in the rate and extent of intracellular accumulation of the nanoparticles between human cancerous and non‐cancerous primary cells, the rate and extent being higher in the malignant cells compared to normal human primary cells. The exposure of the cells to the alkyl‐capped nanocrystals demonstrates no evidence of in vitro cytotoxicity when assessed by cell morphology, apoptosis, and cell viability assays. The internalization of the nanocrystals by Hela and SW1353 cells is almost completely blocked by the pinocytosis inhibitors filipin, cytochalasin B, and actinomycin D. The internalization process is not associated with any surface change in the nanoparticles, as their luminescence spectrum is unaltered upon transport into the cytosol. The observed dramatic difference in the rate and extent of internalization of the nanocrystals between malignant and non‐malignant cells therefore offers potential application in the management of human neoplastic conditions.     

Super‐Efficient Exciton Funneling in Layer‐by‐Layer Semiconductor Nanocrystal Structures

In semiconductor nanocrystals the electronic energy gap is determined not only by the material but also by the size of the nanocrystals. This allows the construction of an energy‐gap gradient normal to multiple layers of nanocrystals where the diameters of the nanocrystals are monotonically increasing or decreasing in subsequent layers. In such devices we observe a highly efficient funneling of excitation energy from layers comprising smaller nanocrystals towards the layer with the largest nanocrystals in the center of the funnel. Most importantly, not only are excitons in radiative states transferred, but also excitons from trapped states, usually lost for luminescence, can be effectively recycled, hence increasing the overall luminescence yield.     

3D Mimicry of Native‐Tissue‐Fiber Architecture Guides Tendon‐Derived Cells and Adipose Stem Cells into Artificial Tendon Constructs

Tendon and ligament (T/L) function is intrinsically related with their unique hierarchically and anisotropically organized extracellular matrix. Their natural healing capacity is, however, limited. Here, continuous and aligned electrospun nanofiber threads (CANT) based on synthetic/natural polymer blends mechanically reinforced with cellulose nanocrystals are produced to replicate the nanoscale collagen fibrils grouped into microscale collagen fibers that compose the native T/L. CANT are then incrementally assembled into 3D hierarchical scaffolds, resulting in woven constructions, which simultaneously mimic T/L nano‐to‐macro architecture, nanotopography, and nonlinear biomechanical behavior. Biological performance is assessed using human‐tendon‐derived cells (hTDCs) and human adipose stem cells (hASCs). Scaffolds nanotopography and microstructure induce a high cytoskeleton elongation and anisotropic organization typical of tendon tissues. Moreover, the expression of tendon‐related markers (Collagen types I and III, Tenascin‐C, and Scleraxis) by both cell types, and the similarities observed on their expression patterns over time suggest that the developed scaffolds not only prevent the phenotypic drift of hTDCs, but also trigger tenogenic differentiation of hASCs. Overall, these results demonstrate a feasible approach for the scalable production of 3D hierarchical scaffolds that exhibit key structural and biomechanical properties, which can be advantageously explored in acellular and cellular T/L TE strategies.     

Worm‐Shape Pt Nanocrystals Grown on Nitrogen‐Doped Low‐Defect Graphene Sheets: Highly Efficient Electrocatalysts for Methanol Oxidation Reaction

Although direct methanol fuel cell offers high energy use efficiency and low pollution emission, the lack of suitable electrode materials poses a great challenge to its commercial application. Herein, a facile and scalable approach is developed to fabricate a hybrid electrocatalyst consisting of strongly coupled worm‐shape Pt nanocrystals and nitrogen‐doped low‐defect graphene (N‐LDG) sheets. Interestingly, it is found that the formation of Pt nanoworms (NWs) is induced by the N atoms in the high‐quality carbon matrix, which also allows the integration of their respective structural advantages and leads to a strong synergetic coupling effect. As a result, the obtained Pt NW/N‐LDG catalyst exhibits an extremely high mass activity of 1283.1 mA mg^?1 toward methanol oxidation reaction, accompanied by reliable long‐term stability and good antipoisoning ability, which are dramatically enhanced as compared with conventional Pt nanoparticle catalysts dispersed on undoped LDG, reduced graphene oxide, and commercial carbon black supports.     

Luminescence Lifetime–Based In Vivo Detection with Responsive Rare Earth–Dye Nanocomposite

For years, luminescence lifetime imaging has served as a quantitative tool in indicating intracellular components and activities. However, very few studies involve the in vivo study of animals, especially in vivo stimuli‐responsive activities of animals, as both excitation and emission wavelengths should fall into the near‐infrared (NIR) optical transparent window (660–950 and 1000–1500 nm). Herein, this work reports a lifetime‐responsive nanocomposite with both excitation and emission in the NIR I window (800 nm) and lifetime in the microsecond region. The incorporation of Tm³⁺‐doped rare‐earth nanocrystals and NIR dye builds an efficient energy transfer pathway that enables a tunable luminescence lifetime range. The NaYF₄:Tm nanocrystal, which absorbs and emits photons at the same energy level, is found to be 33 times brighter than optimized core–shell upconversion nanocrystals, and proved to be an effective donor for NIR luminescence resonance energy transfer (LRET). The anti‐interference capability of luminescence lifetime signals is further confirmed by luminescence and lifetime imaging. In vivo studies also verify the lifetime response upon stimulation generated in an arthritis mouse model. This work introduces an intriguing tool for luminescence lifetime–based sensing in the microsecond region.     

Biomimetic hydroxyapatite coating on glass coverslips for the assay of osteoclast activity in vitro

The osteoclast (OC) is the cell type responsible for the resorption of bone. The activity of this cell is important in the aetiology of a large number of skeletal pathologies, and also for the biocompatibility and osseointegration of orthopaedic implant materials. OC mediated acid hydrolysis of calcium phosphate from the bone matrix offers a prime means of studying the biology and activity of this cell type. We have developed a method of coating glass coverslips with a hydroxyapatite (HA)-like mineral, using a biomimetic approach. Hydroxylation followed by formation of a self assembled monolayer (SAM) using the surfactant triethoxysilylpropyl succinic anhydride (TESPSA), allowed biomimetic deposition of HA-like mineral from a simulated body fluid (SBF). The biocompatibility of the TESPSA SAM-HA coated glass coverslips was tested by culturing human mature OC present in samples of giant cell tumour of bone (GCT). Parameters of OC activity were assayed, including F-actin ring formation, release of calcium and formation of osteoclastic resorption pits, confirming that OC were able to attach to and resorb the coated surface. This approach for the preparation of HA coatings on glass coverslips could have wide applicability for the study of osteoclast behaviour in vitro. Gerald J. Atkins and Peter Majewski share senior author status.     

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 λ_EM‐PL = 436 nm (FeSe) and 452 nm (CuFeSe) while λ_EX‐PL = 356 nm, it also conveys two‐photon phenomenon while λ_EX‐PL = 720 nm. Moreover, it also shows strong off‐resonant luminescence due to two‐photon absorption, which should be valuable for biological applications.     

A Self‐Assembled DNA Origami‐Gold Nanorod Complex for Cancer Theranostics

A self‐assembled DNA origami (DO)‐gold nanorod (GNR) complex, which is a dual‐functional nanotheranostics constructed by decorating GNRs onto the surface of DNA origami, is demonstrated. After 24 h incubation of two structured DO‐GNR complexes with human MCF7 breast cancer cells, significant enhancement of cell uptake is achieved compared to bare GNRs by two‐photon luminescence imaging. Particularly, the triangle shaped DO‐GNR complex exhibits optimal cellular accumulation. Compared to GNRs, improved photothermolysis against tumor cells is accomplished for the triangle DO‐GNR complex by two‐photon laser or NIR laser irradiation. Moreover, the DO‐GNR complex exhibits enhanced antitumor efficacy compared with bare GNRs in nude mice bearing breast tumor xenografts. The results demonstrate that the DO‐GNR complex can achieve optimal two‐photon cell imaging and photothermal effect, suggesting a promising candidate for cancer diagnosis and therapy both in vitro and in vivo.     

Lanthanide‐Doped Core@Multishell Nanoarchitectures: Multimodal Excitable Upconverting/Downshifting Luminescence and High‐Level Anti‐Counterfeiting

The development of luminescent materials with concurrent multimodal emissions is a great challenge to improve security and data storage density. Lanthanide‐doped nanocrystals are particularly appropriate for such applications for their abundant intermediate energy states and distinguishable spectroscopic profiles. However, traditional lanthanide luminescent nanoparticles have a limited capacity for information storage or complexity to shield against counterfeiting. Herein, it is demonstrated that the combination of upconverting and downshifting emissions in a particulate designed lanthanide‐doped core@multishell nanoarchitecture allows the generation of multicolor dual‐modal luminescence over a wide spectral range for complex information storage. Precise control of lanthanide dopants distribution in the core and distinct shells enables simultaneous excitation of 980/808 nm focusing/defocusing laser and 254 nm light and produces complex upconverting emissions from Er, Tm, Eu, and Tb via multiphoton energy transfer processes and downshifting emissions from Eu and Tb via efficient energy transfer from Ce to Eu/Tb in Gd‐assisted lattices. It is experimentally proven that multiple visualized anti‐counterfeit and information encryption with facile decryption and authentication using screen‐printing inks containing the present core@multishell nanocrystals are practically applicable by selecting different excitation modes.     

Recent Progress in Time‐Resolved Biosensing and Bioimaging Based on Lanthanide‐Doped Nanoparticles

Luminescent nanomaterials have attracted great attention in luminescence‐based bioanalysis due to their abundant optical and tunable surface physicochemical properties. However, luminescent nanomaterials often suffer from serious autofluorescence and light scattering interference when applied to complex biological samples. Time‐resolved luminescence methodology can efficiently eliminate autofluorescence and light scattering interference by collecting the luminescence signal of a long‐lived probe after the background signals decays completely. Lanthanides have a unique [Xe]4f^N electronic configuration and ladder‐like energy states, which endow lanthanide‐doped nanoparticles with many desirable optical properties, such as long luminescence lifetimes, large Stokes/anti‐Stokes shifts, and sharp emission bands. Due to their long luminescence lifetimes, lanthanide‐doped nanoparticles are widely used for high‐sensitive biosensing and high‐contrast bioimaging via time‐resolved luminescence methodology. In this review, recent progress in the development of lanthanide‐doped nanoparticles and their application in time‐resolved biosensing and bioimaging are summarized. At the end of this review, the current challenges and perspectives of lanthanide‐doped nanoparticles for time‐resolved bioapplications are discussed.     

Color Patterning of Luminescent Perovskites via Light‐Mediated Halide Exchange with Haloalkanes

Lead halide perovskite possesses a semiconductor bandgap that is readily tunable by a variation in its halide composition. Here, a photo‐activated halide exchange process between perovskite nanocrystals and molecular haloalkanes is reported, which enables the perovskite luminescence to be controllably shifted across the entire visible spectrum. Mechanistic investigations reveal a mutual exchange of halogens between the perovskite crystal surface and a chemisorbed haloalkane, yielding nanocrystals and haloalkanes with mixed halide contents. Exchange kinetics studies involving primary, secondary, and tertiary haloalkanes show that the rate of reaction is governed by the activation barrier in the breakage of the covalent carbon–halogen (C? X) bond, which is a function of the C? X bond energy and carbon radical stability. Employing this halide exchange approach, a micrometer‐scale trichromatic patterning of perovskites is demonstrated using a light‐source‐integrated inkjet printer and tertiary haloalkanes as color‐conversion inks. The haloalkanes volatilize after halide exchange and leave no residues, thereby offering significant processing advantage over conventional salt‐based exchange techniques. Beyond the possible applications in new‐generation micro‐LED and electroluminescent quantum dot displays, this work exemplifies the rich surface and photochemistry of perovskite nanocrystals, and could lead to further opportunities in perovskite‐based photocatalysis and photochemical sensing.     

Size‐Dependent Oxidation of Monodisperse Silicon Nanocrystals with Allylphenylsulfide Surfaces

The synthesis and characterization of size‐separated silicon nanocrystals functionalized with a heteroatom‐substituted organic capping group, allylphenylsulfide, via photochemical hydrosilylation are described for the first time. These silicon nanocrystals form colloidally stable and highly photoluminescent dispersions in non‐polar organic solvents with an absolute quantum yield as high as 52% which is 20% above that of the allylbenzene analogue. Solutions of the size‐separated fractions are characterized over time to monitor the effect of aging in air by following the change of their photoluminescence and absolute quantum yields, supplemented by transmission electron microscopy.     

Ultrafast Single‐Band Upconversion Luminescence in a Liquid‐Quenched Amorphous Matrix

Achieving single‐band upconversion is a challenging but rewarding approach to attain optimal performance in diverse applications, such as multiplexed molecular imaging, security coding, and nonlinear photonic devices. Here, highly efficient single‐band upconversion luminescence in the green spectral regime (16.4 times increase in emission at 525 nm) accomplished by realizing minimal energy loss from two‐photon upconversion in a newly synthesized liquid‐quenched amorphous matrix is reported. In contrast to previously reported single‐band upconversion, this phenomenon originates from the elevated transition probability of the host sensitive transition via changes in the host matrix's microstructure. The elevated transition probability facilitates ultrafast decay of upconversion luminescence with decay times as short as 0.2 µs, the fastest decay ever reported. The material in this study therefore has strong potential for use in photonic devices demanding high upconversion efficiency with a fast response time, which to date has been inaccessible using upconversion materials.     

Toward High‐Dimensional Single‐Cell Analysis of Graphene Oxide Biological Impact: Tracking on Immune Cells by Single‐Cell Mass Cytometry

Considering the potential exposure to graphene, the most investigated nanomaterial, the assessment of the impact on human health has become an urgent need. The deep understanding of nanomaterial safety is today possible by high‐throughput single‐cell technologies. Single‐cell mass cytometry (cytometry by time‐of flight, CyTOF) shows an unparalleled ability to phenotypically and functionally profile complex cellular systems, in particular related to the immune system, as recently also proved for graphene impact. The next challenge is to track the graphene distribution at the single‐cell level. Therefore, graphene oxide (GO) is functionalized with AgInS₂ nanocrystals (GO–In), allowing to trace GO immune–cell interactions via the indium (¹¹⁵In) channel. Indium is specifically chosen to avoid overlaps with the commercial panels (>30 immune markers). As a proof of concept, the GO–In CyTOF tracking is performed at the single‐cell level on blood immune subpopulations, showing the GO interaction with monocytes and B cells, therefore guiding future immune studies. The proposed approach can be applied not only to the immune safety assessment of the multitude of graphene physical and chemical parameters, but also for graphene applications in neuroscience. Moreover, this approach can be translated to other 2D emerging materials and will likely advance the understanding of their toxicology.     

A Single‐Cell Assay for Time Lapse Studies of Exosome Secretion and Cell Behaviors

To understand the inhomogeneity of cells in biological systems, there is a growing demand on the capability of characterizing the properties of individual single cells. Since single‐cell studies require continuous monitoring of the cell behaviors, an effective single‐cell assay that can support time lapsed studies in a high throughput manner is desired. Most currently available single‐cell technologies cannot provide proper environments to sustain cell growth and, proliferation of single cells and convenient, noninvasive tests of single‐cell behaviors from molecular markers. Here, a highly versatile single‐cell assay is presented that can accommodate different cellular types, enable easy and efficient single‐cell loading and culturing, and be suitable for the study of effects of in vitro environmental factors in combination with drug screening. One salient feature of the assay is the noninvasive collection and surveying of single‐cell secretions at different time points, producing unprecedented insight of single‐cell behaviors based on the biomarker signals from individual cells under given perturbations. Above all, the acquired information is quantitative, for example, measured by the number of exosomes each single‐cell secretes for a given time period. Therefore, our single‐cell assay provides a convenient, low‐cost, and enabling tool for quantitative, time lapsed studies of single‐cell properties.     

Near‐Infrared Light‐Sensitive Polyvinyl Alcohol Hydrogel Photoresist for Spatiotemporal Control of Cell‐Instructive 3D Microenvironments

Advanced hydrogel systems that allow precise control of cells and their 3D microenvironments are needed in tissue engineering, disease modeling, and drug screening. Multiphoton lithography (MPL) allows true 3D microfabrication of complex objects, but its biological application requires a cell‐compatible hydrogel resist that is sufficiently photosensitive, cell‐degradable, and permissive to support 3D cell growth. Here, an extremely photosensitive cell‐responsive hydrogel composed of peptide‐crosslinked polyvinyl alcohol (PVA) is designed to expand the biological applications of MPL. PVA hydrogels are formed rapidly by ultraviolet light within 1 min in the presence of cells, providing fully synthetic matrices that are instructive for cell‐matrix remodeling, multicellular morphogenesis, and protease‐mediated cell invasion. By focusing a multiphoton laser into a cell‐laden PVA hydrogel, cell‐instructive extracellular cues are site‐specifically attached to the PVA matrix. Cell invasion is thus precisely guided in 3D with micrometer‐scale spatial resolution. This robust hydrogel enables, for the first time, ultrafast MPL of cell‐responsive synthetic matrices at writing speeds up to 50 mm s^?1. This approach should enable facile photochemical construction and manipulation of 3D cellular microenvironments with unprecedented flexibility and precision.     

Direct Bandgap Silicon: Tensile‐Strained Silicon Nanocrystals

Silicon, a semiconductor underpinning the vast majority of microelectronics, is an indirect‐gap material and consequently is an inefficient light emitter. This hampers the ongoing worldwide effort towards the integration of optoelectronics on silicon wafers. Even though silicon nanocrystals are much better light emitters, they retain the indirect‐gap nature. Here, we propose a solution to this long‐standing problem: silicon nanocrystals can be transformed into a material with fundamental direct bandgap via a concerted action of quantum confinement and tensile strain. We document this transformation by DFT calculations mapping the E( k ) band‐structure of Si nanocrystals. The experimental proofs are then given firstly by a 10 000× increase in the photon emission rate of strained silicon nanocrystals together with their altered absorbance spectra, both of which point to direct dipole‐allowed transitions, secondly by single nanocrystal spectroscopy, confirming reduced phonon energies and thus the presence of tensile strain, and lastly by photoluminescence studies under external hydrostatic pressure.     

Zinc‐Dithizone Complex Engineered Upconverting Nanosensors for the Detection of Hypochlorite in Living Cells

Current chemo/biosensors for hypochlorous acid or hypochlorite detections are usually limited to the submicromolar level because of their insufficient sensitivity, which is a problem because the concentrations in biological matrices is generally on the nanomolar scale or even lower. Developing a probe with a high enough sensitivity remains a challenge. Using the minimal background fluorescence of upconversion nanocrystals to our advantage, we herein report on an energy‐transfer mechanism‐based upconversion luminescent nanosensor for the sensitive and selective detection of hypochlorite in aqueous solution. In this nanosensor water‐dispersible upconversion nanoparticles act as the energy donor and a novel hypochlorite‐responsive coordination complex Zn(DZ)₃ is employed as the energy acceptor. The quenched upconversion luminescence, induced by the Zn(DZ)₃ complex, can be efficiently recovered after addition of hypochlorite through the selective oxidative breakage of the Zn‐S‐C bonds in the Zn(DZ)₃ complex, which was verified by mass spectrometry. The detection limit for hypochlorite of this sensing system is as low as 3 nM. Furthermore, this newly coordination‐complex engineered upconversion nanosensor is successfully applied to image different amounts of exogenous hypochlorite in living HeLa cells.     

Nanobodies and Nanocrystals: Highly Sensitive Quantum Dot‐Based Homogeneous FRET Immunoassay for Serum‐Based EGFR Detection

Semiconductor quantum dot nanocrystals (QDs) for optical biosensing applications often contain thick polyethylene glycol (PEG)‐based coatings in order to retain the advantageous QD properties in biological media such as blood, serum or plasma. On the other hand, the application of QDs in Förster resonance energy transfer (FRET) immunoassays, one of the most sensitive and most common fluorescence‐based techniques for non‐competitive homogeneous biomarker diagnostics, is limited by such thick coatings due to the increased donor‐acceptor distance. In particular, the combination with large IgG antibodies usually leads to distances well beyond the common FRET range of approximately 1 to 10 nm. Herein, time‐gated detection of Tb‐to‐QD FRET for background suppression and an increased FRET range is combined with single domain antibodies (or nanobodies) for a reduced distance in order to realize highly sensitive QD‐based FRET immunoassays. The “(nano)²” immunoassay (combination of nanocrystals and nanobodies) is performed on a commercial clinical fluorescence plate reader and provides sub‐nanomolar (few ng/mL) detection limits of soluble epidermal growth factor receptor (EGFR) in 50 μL buffer or serum samples. Apart from the first demonstration of using nanobodies for FRET‐based immunoassays, the extremely low and clinically relevant detection limits of EGFR demonstrate the direct applicability of the (nano)^2‐ assay to fast and sensitive biomarker detection in clinical diagnostics.     

Bioinspired Diselenide‐Bridged Mesoporous Silica Nanoparticles for Dual‐Responsive Protein Delivery

Controlled delivery of protein therapeutics remains a challenge. Here, the inclusion of diselenide‐bond‐containing organosilica moieties into the framework of silica to fabricate biodegradable mesoporous silica nanoparticles (MSNs) with oxidative and redox dual‐responsiveness is reported. These diselenide‐bridged MSNs can encapsulate cytotoxic RNase A into the 8–10 nm internal pores via electrostatic interaction and release the payload via a matrix‐degradation controlled mechanism upon exposure to oxidative or redox conditions. After surface cloaking with cancer‐cell‐derived membrane fragments, these bioinspired RNase A‐loaded MSNs exhibit homologous targeting and immune‐invasion characteristics inherited from the source cancer cells. The efficient in vitro and in vivo anti‐cancer performance, which includes increased blood circulation time and enhanced tumor accumulation along with low toxicity, suggests that these cell‐membrane‐coated, dual‐responsive degradable MSNs represent a promising platform for the delivery of bio‐macromolecules such as protein and nucleic acid therapeutics.