Vaccine delivery: Nanopatch‐Targeted Skin Vaccination against West Nile Virus and Chikungunya Virus in Mice (Small 16/2010)

The ‘Nanopatch’ (NP) comprises arrays of densely packed projections with a defined geometry and distribution designed to physically target vaccines directly to thousands of epidermal and dermal antigen presenting cells (APCs). These miniaturized arrays are two orders of magnitude smaller than standard needles—which deliver most vaccines—and are also much smaller than current microneedle arrays. The NP is dry‐coated with antigen, adjuvant, and/or DNA payloads. After the NP was pressed onto mouse skin, a protein payload co‐localized with 91.4 ± 4.1 APC mm⁻² (or 2925 in total) representing 52% of the delivery sites within the NP contact area, agreeing well with a probability‐based model used to guide the device design; it then substantially increases as the antigen diffuses in the skin to many more cells. APC co‐localizing with protein payloads rapidly disappears from the application area, suggesting APC migration. The NP also delivers DNA payloads leading to cutaneous expression of encoded proteins within 24 h. The efficiency of NP immunization is demonstrated using an inactivated whole chikungunya virus vaccine and a DNA‐delivered attenuated West Nile virus vaccine. The NP thus offers a needle‐free, versatile, highly effective vaccine delivery system that is potentially inexpensive and simple to use.     

High‐Performance Integrated Enzyme Cascade Bioplatform Based on Protein–BiPt Nanochain@Graphene Oxide Hybrid Guided One‐Pot Self‐Assembly Strategy

Nanozymes provide new opportunities for facilitating next generation artificial enzyme cascade platforms. However, the fabrication of high‐performance integrated artificial enzyme cascade (IAEC) bioplatforms based on nanozymes remains a great challenge. A facile and effective self‐assembly strategy for constructing an IAEC system based on an inorganic/protein hybrid nanozyme, β‐casein‐BiPt nanochain@GO (CA‐BiPtNC@GO) nanohybrid with unique physicochemical surface properties and hierarchical structures, is introduced here. Due to the synergetic effect of the protein, GO, and Bi³⁺, the hybrid acts as highly adaptable building blocks to immobilize natural enzymes directly and noncovalently without the loss of enzyme activity. Simultaneously, the CA‐BiPtNC@GO nanohybrid exhibits outstanding peroxidase‐mimicking activity and works well with natural oxidases, resulting in prominent activity in catalyzing cascade reactions. As a result, the proposed IAEC bioplatform exhibits excellent sensitivity with a wide linear range of 0.5 × 10^‐6 to 100 × 10^‐6 m and a detection limit of 0.05 × 10^‐6 m for glucose. Meticulous design of ingenious hierarchically nanostructured nanozymes with unique physicochemical surface properties can provide a facile and efficient way to immobilize and stabilize nature enzymes using self‐assembly instead of chemical processes, and fill the gap in developing robust nanozyme–triggered IAEC systems with applications in the environment, sensing, and synthetic biology.     

Protein‐Corona‐by‐Design in 2D: A Reliable Platform to Decode Bio–Nano Interactions for the Next‐Generation Quality‐by‐Design Nanomedicines

Hard corona (HC) protein, i.e., the environmental proteins of the biological medium that are bound to a nanosurface, is known to affect the biological fate of a nanomedicine. Due to the size, curvature, and specific surface area (SSA) 3‐factor interactions inherited in the traditional 3D nanoparticle, HC‐dependent bio–nano interactions are often poorly probed and interpreted. Here, the first HC‐by‐design case study in 2D is demonstrated that sequentially and linearly changes the HC quantity using functionalized graphene oxide (GO) nanosheets. The HC quantity and HC quality are analyzed using NanoDrop and label‐free liquid chromatography–mass spectrometry (LC‐MS) followed by principal component analysis (PCA). Cellular responses (uptake and cytotoxicity in J774 cell model) are compared using imaging cytometry and the modified lactate dehydrogenase assays, respectively. Cellular uptake linearly and solely correlates with HC quantity (R² = 0.99634). The nanotoxicity, analyzed by retrospective design of experiment (DoE), is found to be dependent on the nanomaterial uptake (primary), HC composition (secondary), and nanomaterial exposure dose (tertiary). This unique 2D design eliminates the size–curvature–SSA multifactor interactions and can serve as a reliable screening platform to uncover HC‐dependent bio–nano interactions to enable the next‐generation quality‐by‐design (QbD) nanomedicines for better clinical translation.     

Carbon Nanotubes Multifunctionalized by Rolling Circle Amplification and Their Application for Highly Sensitive Detection of Cancer Markers

There are still challenges for the development of multifunctional carbon nanotubes (CNTs). Here, a multiwalled carbon nanotube (MWCNT)‐based rolling circle amplification system (CRCAS) is reported which allows in situ rolling circle replication of DNA primer on the surface of MWCNTs to create a long single‐strand DNA (ssDNA) where a large number of nanoparticles or proteins could be loaded, forming a nano‐biohybridized 3D structure with a powerful signal amplification ability. In this strategy, the binding ability of proteins, hybridization, replication ability of DNA, and the catalytical ability of enzymes are integrated on a single carbon nanotube. The CRCAS is then used to develop colorimetric and chemiluminescent assays for the highly sensitive and specific detection of cancer protein markers, alpha‐fetoprotein (AFP) and prostate specific antigen (PSA). The colorimetric CRCAS assay is 4000 times more sensitive than a conventional enzyme‐linked immunosorbent assay (ELISA), and its concentration range is 10 000 times wider. Control experiments show that as low as 10 pg mL^?1 AFP or PSA could be detected even in the presence of interfering protein markers with a more than 10⁵‐fold greater concentration in the sample, demonstrating the high specificity of the CRCAS assay. The limit of detection of the chemiluminescent CRCAS assays for AFP and PSA are 5 fg mL^?1 (70 aM) and 10 fg mL^?1 (0.29 fM), respectively, indicating that the sensitivity is much higher than that of the colorimetric CRCAS assay. Importantly, CRCAS works well with real biological samples.     

Toward Precise Manipulation of DNA–Protein Hybrid Nanoarchitectures

Nucleic acids and proteins are the two primary building materials of living organisms. Over the past decade, artificial DNA–protein hybrid structures have been pursued for a wide range of applications. DNA nanotechnology, in particular, has dramatically expanded nanoscale molecule engineering and contributed to the spatial arrangement of protein components. Strategies for designing site‐specific coupling of DNA oligomers to proteins are needed in order to allow for precise control over stoichiometry and position. Efforts have also been focused on coassembly of protein–DNA complexes by engineering their fundamental molecular recognition interactions. This Concept focuses on the precise manipulation of DNA–protein nanoarchitectures. Particular attention is paid to site‐selectivity within DNA–protein conjugates, regulation of protein orientation using DNA scaffolds, and coassembly principles upon unique structural motifs. Current challenges and future directions are also discussed in the design and application of DNA–protein nanoarchitectures.     

Recent Advances in Synthesis and Biomedical Applications of Two‐Dimensional Transition Metal Dichalcogenide Nanosheets

During recent decades, a giant leap in the development of nanotechnology has been witnessed. Numerous nanomaterials with different dimensions and unprecedented features have been developed and provided unimaginably wide scope to solve the challenging problems in biomedicine, such as cancer diagnosis and therapy. Recently, two‐dimensional (2D) transition metal dichalcogenide (TMDC) nanosheets (NSs), including MoS₂, WS₂, and etc., have emerged as novel inorganic graphene analogues and attracted tremendous attention due to their unique structures and distinctive properties, and opened up great opportunities for biomedical applications, including ultrasensitive biosensing, biological imaging, drug delivery, cancer therapy, and antibacterial treatment. A comprehensive overview of different synthetic methods of ultrathin 2D TMDC NSs and their state‐of‐the‐art biomedical applications, especially those that have appeared in the past few years, is presented. At the end of this review, the future opportunities and challenges for 2D TMDC NSs in biomedicine are also discussed.     

Dual‐Phase CsPbBr3–CsPb2Br5 Perovskite Thin Films via Vapor Deposition for High‐Performance Rigid and Flexible Photodetectors

Inorganic perovskites with special semiconducting properties and structures have attracted great attention and are regarded as next generation candidates for optoelectronic devices. Herein, using a physical vapor deposition process with a controlled excess of PbBr₂, dual‐phase all‐inorganic perovskite composite CsPbBr₃–CsPb₂Br₅ thin films are prepared as light‐harvesting layers and incorporated in a photodetector (PD). The PD has a high responsivity and detectivity of 0.375 A W^?1 and 10¹¹ Jones, respectively, and a fast response time (from 10% to 90% of the maximum photocurrent) of ≈280 µs/640 µs. The device also shows an excellent stability in air for more than 65 d without encapsulation. Tetragonal CsPb₂Br₅ provides satisfactory passivation to reduce the recombination of the charge carriers, and with its lower free energy, it enhances the stability of the inorganic perovskite devices. Remarkably, the same inorganic perovskite photodetector is also highly flexible and exhibits an exceptional bending performance (>1000 cycles). These results highlight the great potential of dual‐phase inorganic perovskite films in the development of optoelectronic devices, especially for flexible device applications.     

Solid‐Supported Biomolecules on Modified Silica Surfaces—A Tool for Fast Physicochemical Characterization and High‐Throughput Screening

Biofunctionalization for a wide variety of applications can be achieved by coating silica surfaces with biomolecules such as lipids or proteins. However, specific surface optimization of the inorganic SiO₂ is necessary to achieve biocompatible surfaces. Surface shielded porous silica beads can be non‐covalently coated with a single lipid bilayer. The lipids retain their fluidity in this handy solid‐supported system, perfectly mimicking the soft‐surface properties of cellular membranes. A supramolecular architecture can also be used for functional immobilization of membrane proteins: An artificial cytosolic compartment can be created with the aid of polymers; coating by lipid membranes and integration of membrane proteins results in a solid‐supported biofunctional cellular surface. Another surface modification enables a direct immobilization of human serum albumin (HSA) molecules onto silica surfaces. The HSA on this otherwise passivated surface provides a convenient material for the investigation of unspecific protein binding of pharmaceuticals on a high‐throughput scale.     

Repurposing Cyanine NIR‐I Dyes Accelerates Clinical Translation of Near‐Infrared‐II (NIR‐II) Bioimaging

The significantly reduced tissue autofluorescence and scattering in the NIR‐II region (1000–1700 nm) opens many exciting avenues for detailed investigation of biological processes in vivo. However, the existing NIR‐II fluorescent agents, including many molecular dyes and inorganic nanomaterials, are primarily focused on complicated synthesis routes and unknown immunogenic responses with limited potential for clinical translation. Herein, the >1000 nm tail emission of conventional biocompatible NIR cyanine dyes with emission peaks at 700–900 nm is systematically investigated, and a type of bright dye for NIR‐II imaging with high potential for accelerating clinical translation is identified. The asymmetry of the π domain in the S₁ state of NIR cyanine dyes is proven to result in a twisted intramolecular charge‐transfer process and NIR‐II emission, establishing a general rule to guide future NIR‐I/II fluorophore synthesis. The screened NIR dyes are identified to possess a bright emission tail in the NIR‐II region along with high quantum yield, high molar‐extinction coefficient, rapid fecal excretion, and functional groups amenable for bioconjugation. As a result, NIR cyanine dyes can be used for NIR‐II imaging to afford superior contrast and real‐time imaging of several biological models, facilitating the translation of NIR‐II bioimaging to clinical theranostic applications.     

High‐Efficiency Solution‐Processed Inorganic Metal Halide Perovskite Light‐Emitting Diodes

This paper reports highly bright and efficient CsPbBr₃ perovskite light‐emitting diodes (PeLEDs) fabricated by simple one‐step spin‐coating of uniform CsPbBr₃ polycrystalline layers on a self‐organized buffer hole injection layer and stoichiometry‐controlled CsPbBr₃ precursor solutions with an optimized concentration. The PeLEDs have maximum current efficiency of 5.39 cd A^?1 and maximum luminance of 13752 cd m^?2. This paper also investigates the origin of current hysteresis, which can be ascribed to migration of Br^? anions. Temperature dependence of the electroluminescence (EL) spectrum is measured and the origins of decreased spectrum area, spectral blue‐shift, and linewidth broadening are analyzed systematically with the activation energies, and are related with Br^? anion migration, thermal dissociation of excitons, thermal expansion, and electron–phonon interaction. This work provides simple ways to improve the efficiency and brightness of all‐inorganic polycrystalline PeLEDs and improves understanding of temperature‐dependent ion migration and EL properties in inorganic PeLEDs.     

Self‐Assembly of Ferromagnetic Organic–Inorganic Perovskite‐Like Films

Perovskite‐based organic–inorganic hybrids hold great potential as active layers in electronics or optoelectronics or as components of biosensors. However, many of these applications require thin films grown with good control over structure and thickness—a major challenge that needs to be addressed. The work presented here is an effort towards this goal and concerns the layer‐by‐layer deposition at ambient conditions of ferromagnetic organic–inorganic hybrids consisting of alternating CuCl₄‐octahedra and organic layers. The Langmuir‐Blodgett technique used to assemble these structures provides intrinsic control over the molecular organization and film thickness down to the molecular level. Magnetic characterization reveals that the coercive field for these thin films is larger than that for solution‐grown layered bulk crystals. The strategy presented here suggests a promising cost effective route to facilitate the excellently controlled growth of sophisticated materials on a wide variety of substrates that have properties relevant for the high density storage media and spintronic devices.     

Rare‐Earth‐Element‐Ytterbium‐Substituted Lead‐Free Inorganic Perovskite Nanocrystals for Optoelectronic Applications

Lead‐(Pb‐) halide perovskite nanocrystals (NCs) are interesting nanomaterials due to their excellent optical properties, such as narrow‐band emission, high photoluminescence (PL) efficiency, and wide color gamut. However, these NCs have several critical problems, such as the high toxicity of Pb, its tendency to accumulate in the human body, and phase instability. Although Pb‐free metal (Bi, Sn, etc.) halide perovskite NCs have recently been reported as possible alternatives, they exhibit poor optical and electrical properties as well as abundant intrinsic defect sites. For the first time, the synthesis and optical characterization of cesium ytterbium triiodide (CsYbI₃) cubic perovskite NCs with highly uniform size distribution and high crystallinity using a simple hot‐injection method are reported. Strong excitation‐independent emission and high quantum yields for the prepared NCs are verified using photoluminescence measurements. Furthermore, these CsYbI₃ NCs exhibit potential for use in organic–inorganic hybrid photodetectors as a photoactive layer. The as‐prepared samples exhibit clear on–off switching behavior as well as high photoresponsivity (2.4 × 10³ A W^?1) and external quantum efficiency (EQE, 5.8 × 10⁵%) due to effective exciton dissociation and charge transport. These results suggest that CsYbI₃ NCs offer tremendous opportunities in electronic and optoelectronic applications, such as chemical sensors, light emitting diodes (LEDs), and energy conversion and storage devices.     

Liposome Crosslinked Polyacrylamide/DNA Hydrogel: a Smart Controlled‐Release System for Small Molecular Payloads

A novel stimuli‐responsive hydrogel system with liposomes serving as both noncovalent crosslinkers and functional small molecules carriers for controlled‐release is developed. Liposomes can crosslink polyacrylamide copolymers functionalized with cholesterol‐modified DNA motifs to yield a DNA hydrogel system, due to the hydrophobic interaction between cholesteryl groups and the lipid bilayer of liposomes. Functional information encoded DNA motifs on the polymer backbones endow the hydrogel with programmable smart responsive properties. In a model system, the hydrogel exhibits stimuli‐responsive gel‐to‐sol transformation triggered by the opening of DNA motifs upon the presence of a restriction endonuclease enzyme, EcoR I, or temperature change, realizing the controlled‐release of liposomes which are highly efficient carriers of active small molecules payloads. Two active molecules, 1,1‐dioctadecyl‐3,3,3,3‐tetramethylindodicarbocyanine perchlorate (DiIC18(5)) and calcein, are chosen as the hydrophobic and hydrophilic model payloads, respectively, to address the feasibility of the releasing strategy. Moreover, the hydrogel exhibits injectable property as well as self‐recovery behaviors.     

Proton‐Functionalized Two‐Dimensional Graphitic Carbon Nitride Nanosheet: An Excellent Metal‐/Label‐Free Biosensing Platform

Ultrathin graphitic carbon nitride (g‐C₃N₄) nanosheets, due to their interesting two‐dimensional graphene‐like structure and unique physicochemical properties, have attracted great research attention recently. Here, a new approachis developed to prepare, for the first time, proton‐functionalized ultrathin g‐C₃N₄ nanosheets by sonication‐exfoliation of bulk g‐C₃N₄ under an acid condition. This method not only reduces the exfoliation time from more than 10 h to 2 h, but also endows the nanosheets with positive charges. Besides retaining the properties of g‐C₃N₄, the obtained nanosheets with the thickness of 2–4 nm (i.e., 6–12 atomic monolayers) also exhibit large specific surface area of 305 m² g^?1, enhanced fluorescence intensity, and excellent water dispersion stability due to their surface protonation and ultrathin morphology. The well‐dispersed protonated g‐C₃N₄ nanosheets are able to interact with negatively charged heparin, which results in the quenching of g‐C₃N₄ fluorescence. A highly sensitive and highly selective heparin sensing platform based on protonated g‐C₃N₄ nanosheets is established. This metal‐free and fluorophore label‐free system can reach the lowest heparin detection limit of 18 ng mL^?1.     

Quantification of Uptake and Localization of Bovine Serum Albumin‐Stabilized Single‐Wall Carbon Nanotubes in Different Human Cell Types

Single‐wall carbon nanotubes (SWCNTs) possess many unique, inherent properties that make them attractive materials for application in medical and biological technologies. Development of concentrated SWCNT dispersions of isolated nanotubes that retain SWCNTs' inherent properties with minimal negative cellular effects is essential to fully realize the potential of SWCNTs in biotechnology. It is shown that bovine serum albumin (BSA), a common and well‐characterized model blood serum protein, can individually disperse SWCNTs at concentrations of up to 0.3 mg mL^?1 while retaining SWCNTs' optical properties. Uptake into human mesenchymal stem cells (hMSC) and HeLa cells is quantified, revealing strikingly high concentrations of 86 ± 33 × 10⁶ and 21 ± 13 × 10⁶ SWCNTs per cell, respectively, without any apparent acute deleterious cellular effects. Through high‐resolution confocal Raman spectroscopy and imaging, it is established that SWCNT–BSAs are preferentially localized intracellularly, especially in the cytoplasm of both hMSCs and HeLa cells. The uptake and localization results demonstrate the efficacy of BSA as a biocompatible dispersant and a mediator of bioactivity. BSA is widely available and inexpensive, which make these concentrated, highly‐dispersed, noncovalently modified SWCNT–BSAs suitable for the development of SWCNT‐based biotechnologies.     

Ligand‐Free Fe3O4/CMCS Nanoclusters with Negative Charges for Efficient Structure‐Selective Protein Adsorption

The easy and effective capture of a single protein from a complex mixture is of great significance in proteomics and diagnostics. However, adsorbing nanomaterials are commonly decorated with specific ligands through a complicated and arduous process. Fe₃O₄/carboxymethylated chitosan (Fe₃O₄/CMCS) nanoclusters are developed as a new nonligand modified strategy to selectively capture bovine hemoglogin (BHB) and other structurally similar proteins (i.e., lysozyme (LYZ) and chymotrypsin (CTP)). The ligand‐free Fe₃O₄/CMCS nanoclusters, in addition to their simple and economical two‐step preparation process, possess many merits, including uniform morphology, high negative charges (?27 mV), high saturation magnetization (60 emu g^?1), and high magnetic content (85%). Additionally, the ligand‐free Fe₃O₄/CMCS nanoclusters are found to selectively capture BHB in a model protein mixture even within biological samples. The reason for selective protein capture is further investigated from nanomaterials and protein structure. In terms of nanomaterials, it is found that high negative charges are conducive to selectively adsorb BHB. In consideration of protein structure, interestingly, the ligand‐free magnetic nanoclusters display a structure‐selective protein adsorption capacity to efficiently capture other proteins structurally similar to BHB, such as LYZ and CTP, showing great potential of the ligand‐free strategy in biomedical field.     

In situ Visualization of Gene Expression Using Polymer‐Coated Quantum–Dot–DNA Conjugates

Imaging of specific mRNA targets in cells is of great importance in understanding gene expression and cell signaling processes. Subcellular localization of mRNA is known as a universal mechanism for cells to sequester specific mRNA for high production of required proteins. Various gene expressions in Drosophila cells are studied using quantum dots (QDs) and the fluorescence in situ hybridization (FISH) method. The excellent photostability and highly luminescent properties of QDs compared to conventional fluorophores allows reproducible obtainment of quantifiable mRNA gene expression imaging. Amine‐modified oligonucleotide probes are designed and covalently attached to the carboxyl‐terminated polymer‐coated QDs via EDC chemistry. The resulting QD–DNA conjugates show sequence‐specific hybridization with target mRNAs. Quantitative analysis of FISH on the Diptericin gene after lipopolysaccharide (LPS) treatment shows that the intensity and number of FISH signals per cell depends on the concentration of LPS and correlates well with quantitative real‐time PCR results. In addition, our QD–DNA probes exhibit excellent sensitivity to detect the low‐expressing Dorsal‐related immunity factor gene. Importantly, multiplex FISH of Ribosomal protein 49 and Actin 5C using green and red QD–DNA conjugates allows the observation of cellular distribution of the two independent genes simultaneously. These results demonstrate that highly fluorescent and stable QD–DNA probes can be a powerful tool for direct localization and quantification of gene expression in situ.     

Ordered Arrangement and Optical Properties of Silica‐Stabilized Gold Nanoparticle–PNIPAM Core–Satellite Clusters for Sensitive Raman Detection

Gold–polymer hybrid nanoparticles attract wide interest as building blocks for the engineering of photonic materials and plasmonic (active) metamaterials with unique optical properties. In particular, the coupling of the localized surface plasmon resonances of individual metal nanostructures in the presence of nanometric gaps can generate highly enhanced and confined electromagnetic fields, which are frequently exploited for metal‐enhanced light–matter interactions. The optical properties of plasmonic structures can be tuned over a wide range of properties by means of their geometry and the size of the inserted nanoparticles as well as by the degree of order upon assembly into 1D, 2D, or 3D structures. Here, the synthesis of silica‐stabilized gold–poly(N‐isopropylacrylamide) (SiO₂‐Au‐PNIPAM) core–satellite superclusters with a narrow size distribution and their incorporation into ordered self‐organized 3D assemblies are reported. Significant alterations of the plasmon resonance are found for different assembled structures as well as strongly enhanced Raman signatures are observed. In a series of experiments, the origin of the highly enhanced signals can be assigned to the interlock areas of adjacent SiO₂‐Au‐PNIPAM core–satellite clusters and their application for highly sensitive nanoparticle‐enhanced Raman spectroscopy is demonstrated.     

Photocatalytic Activity of Protein‐Conjugated CdS Nanoparticles

Colloidal CdS nanoparticles are conjugated with a variety of proteins, including enhanced yellow fluorescent protein, tobacco etch virus protease (TEV), lysozyme, and bacterial cytochrome P450 CYP152A1, and the photochemical properties of the resulting conjugates are analyzed by EPR spectroscopy and hydroxyl radical‐specific fluorimetric assay. While irradiation of bare CdS colloids leads to photogeneration of hydroxyl and superoxide radicals, it is surprisingly observed that coating of the CdS particles with proteins effectively suppresses the production of these radical species and instead leads to increased formation of a long‐lived reactive oxygen species, most likely H₂O₂. A mechanism for the observed results is suggested. The empirical results are capitalized on for the assembly of a CdS–TEV nanohybrid, which shows significantly higher performance as a photocatalytic mediator for fatty acid hydroxylation by CYP152A1 than bare CdS nanoparticles.