Advances in the Application of Microneedles in the Treatment of Local Organ Diseases.

In recent years, microneedles (MNs) have attracted a lot of attention due to their microscale sizes and high surface area (500-1000 µm in length), allowing pain-free and efficient drug delivery through the skin. In addition to the great success of MNs based transdermal drug delivery, especially for skin diseases, increasing studies have indicated the expansion of MNs to diverse nontransdermal applications, including the delivery of therapeutics for hair loss, ocular diseases, and oral mucosal. Here, the current treatment of hair loss, eye diseases, and oral disease is discussed and an overview of recent advances in the application of MNs is provided for these three noncutaneous localized organ diseases. Particular emphasis is laid on the future trend of MNs technology development and future challenges of expanding the generalizability of MNs.

Multiplexed Organelles Portrait Barcodes for Subcellular MicroRNA Array Detection in Living Cells.

Multiplexed profiling of microRNAs' subcellular expression and distribution is essential to understand their spatiotemporal function information, but it remains a crucial challenge. Herein, we report an encoding approach that leverages combinational fluorescent dye barcodes, organelle targeting elements, and an independent quantification signal, termed Multiplexed Organelles Portrait Barcodes (MOPB), for high-throughput profiling of miRNAs from organelles. The MOPB barcodes consist of heterochromatic fluorescent dye-loaded shell-core mesoporous silica nanoparticles modified with organelle targeting peptides and molecular beacon detection probes. Using mitochondria and endoplasmic reticulum as models, we encoded four Cy3/AMCA ER-MOPB and four Cy5/AMCA Mito-MOPB by varying the Cy3 and Cy5 intensity for distinguishing eight organelles' miRNAs. Significantly, the MOPB strategy successfully and accurately profiled eight subcellular organelle miRNAs' alterations in the drug-induced Ca2+ homeostasis breakdown. The approach should allow more widespread application of subcellular miRNAs and multiplexed subcellular protein biomarkers' monitoring for drug discovery, cellular metabolism, signaling transduction, and gene expression regulation readout.

Metal-Organic Framework-Loaded Engineering DNAzyme for the Self-Powered Amplified Detection of MicroRNA.

DNAzyme shows great promise in designing a highly sensitive and specific sensing platform; however, the low cellular uptake efficiency, instability, and especially the insufficient cofactor supply inhibit the intracellular molecule sensor applications. Herein, we demonstrate a novel type of DNAzyme-based self-driven intracellular sensor for microRNA (miRNA) detection in living cells. The sensor consists of a metal-organic framework [zeolite imidazole framework (ZIF-8)] core loaded with a shell consisting of a rationally designed DNAzyme, where the substrate strand is modified with FAM and BHQ-1 nearby both the sides of the restriction site, respectively, while the enzyme strand consists of two separate strands with a complementary fragment to the substrate strand and the targeting miRNA, respectively. The ZIF-8 nanoparticles enable the efficient delivery of DNAzyme into the cell and protect the DNAzyme from degradation. The pH-responsive ZIF-8 degradation is accompanied with the release of the DNAzyme and Zn2+ cofactors, and the intracellular target miRNAs recognize and activate the DNAzyme driven by the Zn2+ cofactors to cleave the substrate strand, resulting in the release of the FAM-labeled shorter product strand and increased fluorescence for miRNA detection. The self-driven approach can be generally applied to various miRNAs' detection through DNAzyme engineering.

Engineering DNA walkers for bioanalysis: A review.

Considerable advances have been made in the design, modularization, functionalization, and regulation of DNA nanostructures over the past 40 years. These advances have accelerated the development of DNA nanomachines such as DNA walkers, dynamic nanomachines with walking feet, tracks, and driven forces, which have highly sensitive detection and signal amplification abilities that can be applied to various bioanalytical contexts and therapeutic strategies. Here, we describe a rational design of the nano-bio interface, the kinetics of DNA walkers and the strategies for improving their efficiency and sensitivity. We also outline the various bioanalytic and imaging applications to which DNA walkers have been applied, such as electrochemical and optical measurements, when integrated with other simulation and activation tools. Finally, we compare the performances of novel DNA walker-based strategies for bioanalysis and propose a method to improve DNA walker design.

Microneedle-Based Device for Biological Analysis.

The collection and analysis of biological samples are an effective means of disease diagnosis and treatment. Blood sampling is a traditional approach in biological analysis. However, the blood sampling approach inevitably relies on invasive techniques and is usually performed by a professional. The microneedle (MN)-based devices have gained increasing attention due to their noninvasive manner compared to the traditional blood-based analysis method. In the present review, we introduce the materials for fabrication of MNs. We categorize MN-based devices based on four classes: MNs for transdermal sampling, biomarker capture, detecting or monitoring analytes, and bio-signal recording. Their design strategies and corresponding application are highlighted and discussed in detail. Finally, future perspectives of MN-based devices are discussed.

Endogenous MicroRNA Accurate Diagnostics to Guide Photothermal Therapy.

Developing an intelligent theranostic nanoplatform with satisfied diagnostic accuracy and therapeutic efficiency holds great promise for personalized nanomedicine. Herein, we constructed a smart nanodevice for the accurate diagnosis of endogenous cancer microRNA (miRNA) biomarkers and efficient photothermal therapy (PTT). The nanodevice was composed of polydopamine (PDA)-functionalized CuS nanosheets (CuS@PDA NSs) and three elaborate DNA hairpin probes (TDHPs). The CuS@PDA NSs acted as efficient delivery vehicles and photothermal agents. They provided a large surface area available for an efficient and facile loading of TDHPs and a high-fluorescence (FL) quenching performance to achieve an ultralow background signal. The intracellular miRNA triggered TDHPs to assemble into three-arm branched junction structures for a strong fluorescence recovery as output signals to discriminate cancer cells from normal cells with an excellent sensitivity. The CuS@PAD NSs showed a good photothermal conversion efficiency in the near-infrared II (NIR II) region to mediate a good photothermal performance to kill cancer cells. A remarkable antitumor therapeutic effect was achieved in vivo. This work integrated highly sensitive detection to endogenous cancer biomarkers and valid therapeutic potency to tumor-bearing mice, indicating its promising biomedical applications.

A Sample and Detection Microneedle Patch for Psoriasis MicroRNA Biomarker Analysis in Interstitial Fluid.

Skin interstitial fluid (ISF) containing a great variety of molecular biomarkers derived from cells and subcutaneous blood capillaries has recently emerged as a clinically potential component for early diagnosis of a wide range of diseases; however, the minimally invasive sampling and detection of cell-free biomarkers in ISF is still a key challenge. Herein, we developed microneedles (MNs) that consist of gelatin methacryloyl (GelMA) and graphene oxide (GO) for the enrichment and sensitive detection of multiple microRNA (miRNA) biomarkers from skin ISF. The GO-GelMA MNs exhibited robust mechanical properties, fast sampling kinetics, and large swelling capacity, which enabled collecting ISF volume high to 21.34 µL in 30 min, facilitating effective miRNA analysis. It preliminarily realized the sensitive detection of three types of psoriasis-related miRNAs biomarkers either on the patch itself or in solution after release from the hydrogel by combining catalytic hairpin assembly signal amplification reaction. The automated and minimally invasive ISF miRNA detection technology of GO-GelMA MNs has great potential to monitor cell-free clinically informative biomarkers for personalized diagnosis and prognosis.

Artificial photoactive chlorophyll conjugated vanadium carbide nanostructure for synergistic photothermal/photodynamic therapy of cancer.

Optically active nanostructures consisting of organic compounds and metallic support have shown great promise in phototherapy due to their increased light absorption capacity and high energy conversion. Herein, we conjugated chlorophyll (Chl) to vanadium carbide (V2C) nanosheets for combined photodynamic/photothermal therapy (PDT/PTT), which reserves the advantages of each modality while minimizing the side effects to achieve an improved therapeutic effect. In this system, the Chl from Leptolyngbya JSC-1 extracts acted as an efficient light-harvest antenna in a wide NIR range and photosensitizers (PSs) for oxygen self-generation hypoxia-relief PDT. The available large surface of two-dimensional (2D) V2C showed high Chl loading efficiency, and the interaction between organic Chl and metallic V2C led to energy conversion efficiency high to 78%. Thus, the Chl/ V2C nanostructure showed advanced performance in vitro cell line killing and completely ablated tumors in vivo with 100% survival rate under a single NIR irradiation. Our results suggest that the artificial optical Chl/V2C nanostructure will benefit photocatalytic tumor eradication clinic application.

Spatially Resolved, Error-Robust Multiplexed MicroRNA Profiling in Single Living Cells.

Simultaneous imaging of multiple microRNAs (miRNAs) in individual living cells is challenging due to the lack of spectrally distinct encoded fluorophores and non-cytotoxic methods. We describe a multiplexed error-robust combinatorial fluorescent label-encoding method, termed fluorophores encoded error-corrected labels (FluoELs), enabling multiplexed miRNA imaging in living cells with error-correcting capability. The FluoELs comprise proportional dual fluorophores for encoding and a constant quantitative single fluorophore for error-corrected quantification. Both are embedded in 260 nm core-shell silica nanoparticles modified with molecular beacon detection probes. The FluoELs are low cytotoxic and could accurately quantify and spatially resolve nine breast-cancer-related miRNAs and evaluate their coordination. The FluoELs enabled a single-cell analysis platform to evaluate miRNA expression profiles and the molecular mechanisms underlying miRNA-associated diseases.

Shedding Light on DNA-Based Nanoprobes for Live-Cell MicroRNA Imaging.

DNA-based nanoprobes integrated with various imaging signals have been employed for fabricating versatile biosensor platforms for the study of intracellular biological process and biomarker detection. The nanoprobes developments also provide opportunities for endogenous microRNA (miRNA) in situ analysis. In this review, the authors are primarily interested in various DNA-based nanoprobes for miRNA biosensors and declare strategies to reveal how to customize the desired nanoplatforms. Initially, various delivery vehicles for nanoprobe architectures transmembrane transport are delineated, and their biosecurity and ability for resisting the complex cellular environment are evaluated. Then, the novel strategies for designing DNA sequences as target miRNA specific recognition and signal amplification modules for miRNA detection are presented. Afterward, recent advances in imaging technologies to accurately respond and produce significant signal output are summarized. Finally, the challenges and future directions in the field are discussed.

Multiple amplified microRNAs monitoring in living cells based on fluorescence quenching of Mo2B and hybridization chain reaction.

Imaging intracellular microRNAs (miRNAs) demonstrated an essential role in exposing their biological and pathological functions. However, the detection of sequence-specific miRNAs in living cells remains a key challenge. Herein, a facile amplified multiple intracellular miRNAs imaging platform was constructed based on Mo2B nanosheets (NSs) fluorescence (FL) quenching and hybridization chain reaction (HCR). The Mo2B NSs demonstrated strong interaction with the hairpin probes (HPs), ssDNA loop, and excellent multiple FL dyes quenching performance, achieving ultralow background signal. After transfection, the HPs recognized specific targets miRNAs, the corresponding HCR was triggered to produce tremendous DNA-miRNA duplex helixes, which dissociated from the surface of the Mo2B NSs to produce strong FL for miRNAs detection. It realized to image multiple miRNAs biomarkers in different cells to discriminate cancer cells from normal cells owing to the excellent sensitivity, and the regulated expression change of miRNAs in cancer cells was also successfully monitored. The facile and versatile Mo2B-based FL quenching platform open an avenue to profile miRNAs expression pattern in living cells, and has great applications in miRNAs based biological and biomedical research.

Interstitial Fluid Biomarkers' Minimally Invasive Monitoring Using Microneedle Sensor Arrays.

Skin interstitial fluid (ISF) is a biofluid with information-rich biomarkers for disease diagnosis and prognosis. Microneedle (MN) integration of sampling and instant biomarker readout hold great potential in health status monitoring and point-of-care testing (POCT). The present work describes an attractive MN sensor array for minimally invasive monitoring of ISF microRNA (miRNA) and Cu2+. The MN array is made of methacrylated gelatin (GelMA) and methacrylated hyaluronic acid (MeHA), and a further divisionally encapsulated miRNA and Cu2+ detection system, and is cross-linked through blue-light irradiation. The MN patch displays good mechanical properties that enable withstanding more than 0.4 N per needle, and exhibits a high swelling ratio of 700% that facilitates timely extraction of sufficient ISF for biomarker analysis. For proof-of-concept, it realizes detection of miRNAs and Cu2+ efficiently and quantitatively in an agarose skin and fresh porcine cadaver skin model. Given the good sampling and in situ monitoring ability, the MN array holds great promise for skin ISF-based applications.

DNA Logic Circuits for Multiple Tumor Cells Identification Using Intracellular MicroRNA Molecular Bispecific Recognition.

The aberrant expression level of intracellular microRNAs (miRNAs) holds great promise for differentiating cell types at the molecular level. However, cell subtype discrimination based on a single miRNA molecular level is not sufficient and reliable. Herein, multiple identifiable and reporting modules are integrated into a single DNA circuit for multiple cancer cell subtypes identification based on miRNAs bispecific recognition. The DNA three-dimensional (3D) logic gate nano-hexahedron framework extends three recognition modules and three reporting modules to form three "AND" logic gates. Each Boolean operator "AND" returns an "ON" signal in the presence of bispecific miRNAs, simultaneously enabling three types of cell subtype identification. It not only enables the discrimination of cancer cells A549 and MCF-7 from normal cells NHDF but also successfully distinguishes different types of cancer cells. The bispecific intracellular miRNA controllable DNA circuit, with low signal-to-noise ratio, easily extends to various cell type discrimination by adjusting the miRNA species, provides huge opportunities for accurately differentiating multiple cell types at the molecular level.

Recent progress in near-infrared photoacoustic imaging.

The emergence of the photoacoustic imaging (PAI) expands the application of biomolecules bioimaging in cells, various tissues, and living body to monitor multiple physiological processes in complex internal environments. The PAI possesses intriguing properties such as non-invasive, highly selective excitation, and weak signal attenuation. Especially, the near-infrared (NIR) PAI displays low optical absorption and scattering, good temporal or spatial resolution and deep penetration, holds great potential in biomedical applications. We briefly compare different imaging modalities to provide a comprehensive understanding of their characteristics and related applications, highlighting the feature of the PAI. The principle of PAI is then delineated and the emerging NIR-PAI is discussed. We then focus on elaboration of the recent achievement of typical NIR-PAI contrast and their biomedical applications, especially the strategies used to improve contrast rational design and PAI performance are summarized. The PAI-related multimodal imaging approaches for improving imaging accuracy are also covered in the review. Finally, the challenges and prospective are pointed out for attracting more researchers to accelerate the development of PAI.

Biodegradable FeWOx nanoparticles for CT/MR imaging-guided synergistic photothermal, photodynamic, and chemodynamic therapy.

The development of a simple and effective single constituent multifunctional nanotheranostic platform producing a multimodality diagnostic signal and curing effect is still a challenge. Herein, we synthesized simple and biodegradable FeWOx ternary oxide nanoparticles and modified their surface with RGD-PEG-NH2 (FeWOx-PEG-RGD NPs), whereby resulting NPs possessed a (T2/T1) switchable MRI/CT dual-modal imaging ability and synergistic photothermal therapy (PTT)/photodynamic therapy (PDT)/chemodynamic therapy (CDT) capacity. We showed that FeWOx-PEG-RGD NPs enabled tumor accumulation under a magnetic field drive and RGD-mediated tumor penetration and implemented PTT/PDT treatment under 980 nm laser irradiation. In an acidic tumor microenvironment (TME) with a high hydrogen peroxide (H2O2) expression, NPs degraded to release Fe3+ and Fe2+, triggering a Fenton reaction to generate ˙OH for CDT. The released Fe2+ led to T2/T1 signal conversion for tracing cancer therapy, while the high X-ray attenuation coefficient of W also made it a good CT contrast agent for guided therapy. Thus, the structurally simple FeWOx-PEG-RGD was capable of mediating (T2/T1-weighted) MR/CT two modal imaging-guided PTT/PDT/CDT synergic therapy with a high accuracy and superb anticancer efficiency. The simple, degradable, rapid-clearance, and multifunctional FeWOx-PEG-RGD NPs provide a novel, promising, and versatile nanotheranostic platform.

V2C Nanosheets as Dual-Functional Antibacterial Agents.

Antibiotic-resistant bacterial strains have been continuously increasing and becoming a supreme threat to public health globally. The nanoparticle-based photothermal treatment has emerged as a powerful tool to combat toxic bacteria. Photothermal agents (PTAs) with cost-effective and high photothermal conversion efficiency are highly desirable. Herein, we unite the green process for delamination of V2AlC to produce a high yield mass of two-dimensional (2D) V2C nanosheets (NSs) by using algae extracts and demonstrate their high antibacterial efficiency. The resultant V2C NSs present decent structural reliability and intrinsic antibacterial ability. Powerful near-infrared (NIR) absorption and extraordinary photothermal conversion proficiency make it a good PTA for the photothermal treatment of bacteria. The antibacterial efficiency evaluation indicated that V2C NSs could effectively kill both Gram-positive S. aureus and Gram-negative E. coli. About 99.5% of both types of bacteria could be killed with low-dose of V2C NSs suspension (40 µg/mL) with 5 min NIR irradiation due to the intrinsic antibacterial ability and photothermal effect of V2C NSs, which is much higher than previous reports on Ta4C3, Ti3C2, MoSe2, and Nb2C. This work expands the application of MXene V2C NSs for rapid bacteria-killing and would gain promising attention for applications in the sterilization industry.

Biosorption of iron ions through microalgae from wastewater and soil: Optimization and comparative study.

Microalgae play a significant role in wastewater and soil-bioremediation due to their low-cost and eco-friendly nature. In this study, 21 strains of microalgae were evaluated during removal of iron Fe2+ from aqueous solutions. Out of 21 strains, five strains (S. obliquus, C. fusca, C. saccharophila, A. braunii, and Leptolyngbya JSC-1) were selected based on their comparative tolerance for the iron Fe2+. These strains were further studied for their Fe2+ removal efficiency. The results indicated that the selected strains could maintain normal growth pattern up to 50 ppm of Fe2+, while the concentration beyond 50 ppm inhibited the growth. The Fe2+ bio-removal efficiencies from wastewater were 97, 98, 97.5, 99, and 99.9%, respectively. Similarly, in soil the bio-removal efficiencies of the five strains were measured as 76, 77, 76, 77.5, and 79%, repectively. A slight increase in leakage of protein and nucleic acids was observed in all strains, which is unlikely could be the reason of iron exposure as similar pattern was also found in control groups. Current results suggested that the selected five strains have high potential to be used as bioremediation tools for Fe2+ contaminated water and soil.

Uniform palladium nanosheets for fluorimetric detection of circulating tumor DNA.

Fluorescence quenching property of two-dimensional (2D) nanosheets (NSs) have received extensively attention in the construction of novel biosensing platform. However, the heterogeneity of the wide-size distribution and inefficient fluorescence quenching capacity limit its wide practical applications. Herein, for the first time, we report a novel fluorescent biosensor based on uniform palladium NSs (Pd NSs) with excellent fluorescence quenching efficiency and differential affinity toward ssDNA versus dsDNA and combination with a pair of DNA detection probes with fluorophore for detecting circulating tumor DNA (ctDNA). The DNA detection probes are facilitated to adsorbed to the surface of Pd NSs, leading to efficient fluorescence quench. In the presence of target DNA, it can be linked by T4 DNA ligase to form long DNA duplex structures, which display weak affinity toward Pd NSs, producing the fluorescence recovery. The remarkable fluorescence quenching efficiency and ssDNA/dsDNA differential affinity of Pd NSs make it have a good detection ability without signal amplification. The result indicates that this facile but cost-effective strategy holds great promise in bioanalysis.