Charge-Reversal NaCl/G-Quartets for Aggregation-Induced Mitochondrial MicroRNA Imaging and Ion-Interference Therapy.

Mitochondrial therapy is a promising new strategy that offers the potential to achieve precise disease diagnosis or maximum therapeutic response. However, versatile mitochondrial theranostic platforms that integrate biomarker detection and therapy have rarely been exploited. Here, we report a charge-reversal nanomedicine activated by an acidic microenvironment for mitochondrial microRNA (mitomiR) detection and ion-interference therapy. The transporter liposome (DD-DC) was constructed from a pH-responsive polymer and a positively charged phospholipid, encapsulating NaCl nanoparticles with coloading of the aggregation-induced emission (AIE) fluorogens AIEgen-DNA/G-quadruplexes precursor and brequinar (NAB@DD-DC). The negatively charged nanomedicine ensured good blood stability and high tumor accumulation, while the charge-reversal to positive in response to the acidic pH in the tumor microenvironment (TME) and lysosomes enhanced the uptake by tumor cells and lysosome escape, achieving accumulation in mitochondria. The subsequently released Na+ in mitochondria not only contributed to the formation of mitomiR-494 induced G-quadruplexes for AIE imaging diagnosis but also led to an osmolarity surge that was enhanced by brequinar to achieve effective ion-interference therapy.

Separable Microneedle for Integrated Hyperglycemia Sensing and Photothermal Responsive Metformin Release.

Microdevices that offer hyperglycemia monitoring and controllable drug delivery are urgently needed for daily diabetes management. Herein, a theranostic separable double-layer microneedle (DLMN) patch consisting of a swellable GelMA supporting base layer for glycemia sensing and a phase-change material (PCM) arrowhead layer for hyperglycemia regulation has been fabricated. The Cu-TCPP(Fe)/glucose oxidase composite and 3,3',5,5'-tetramethylbenzidine coembedded in the supporting base layer permit a visible color shift at the base surface in the presence of glucose via a cascade reaction, allowing for the in situ detection of glucose in interstitial fluid. The PCM arrowhead layer is encapsulated with water monodispersity melanin nanoparticles from Sepia officinalis and metformin that is imparted with a near-infrared ray photothermal response feature, which is beneficial to the controllable release of metformin for suppression of hyperglycemia. By applying the DLMN patch to the streptozotocin-induced type 2 diabetic Sprague-Dawley rat model, the results demonstrated that it can effectively extract dermal interstitial fluid, read out glucose levels, and regulate hyperglycemia. This DLMN-integrated portable colorimetric sensor and self-regulated glucose level hold great promise for daily diabetes management.

RNA-Cleaving DNAzyme-Based Amplification Strategies for Biosensing and Therapy.

Since their first discovery in 1994, DNAzymes have been extensively applied in biosensing and therapy that act as recognition elements and signal generators with the outstanding properties of good stability, simple synthesis, and high sensitivity. One subset, RNA-cleaving DNAzymes, is widely employed for diverse applications, including as reporters capable of transmitting detectable signals. In this review, the recent advances of RNA-cleaving DNAzyme-based amplification strategies in scaled-up biosensing are focused, the application in diagnosis and disease treatment are also discussed. Two major types of RNA-cleaving DNAzyme-based amplification strategies are highlighted, namely direct response amplification strategies and combinational response amplification strategies. The direct response amplification strategies refer to those based on novel designed single-stranded DNAzyme, and the combinational response amplification strategies mainly include two-part assembled DNAzyme, cascade reactions, CHA/HCR/RCA, DNA walker, CRISPR-Cas12a and aptamer. Finally, the current status of DNAzymes, the challenges, and the prospects of DNAzyme-based biosensors are presented.

Colorimetric-photothermal-magnetic three-in-one lateral flow immunoassay for two formats of biogenic amines sensitive and reliable quantification.

Rapid, simple, sensitive and reliable approaches for biogenic amines quantification in various food samples are essential to food safety. Lateral flow immunoassay (LFIA) has been wildly utilized in point-of-care testing (POCT) owing to its advantage of flexibility and feasibility. Here, we reported a Fe3O4@Au nanoparticles (NPs) (Fe3O4@AuNPs) based multimodal readout LFIA for rapid putrescine (Put) and histamine (His) quantification with a LOD down to 10 and 10 ng/mL in naked eye mode, 2.31 and 4.39 ng/mL in photothermal mode, 0.17 and 0.31 ng/mL in magnetic mode, respectively. Such multi-mode assay has been successfully used to detect Biogenic amines (BAs) in raw aquatic foods, including fish, prawns, beef, and pork, with overall recoveries ranging from 93.68 to 109.34%. Meanwhile, it is easily expanded to detect other typical BAs with high sensitivity by simply replacing antibodies. In view of the multi-signal reading, two quantitative formats, and high sensitivity, it may greatly widen the application of lateral flow detection in food safety.

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.

Mitochondria MicroRNA Spatial Imaging via pH-Responsive Exonuclease-Assisted AIE Nanoreporter.

Mitochondrial microRNAs (mitomiRs) critically orchestrate mitochondrial functions. Spatial imaging of mitomiRs is essential to understand its clinical value in diagnosis and prognosis. However, the direct monitoring of mitomiRs in living cells remains a key challenge. Herein, we report an AIE nanoreporter strategy for mitomiRs imaging in living cells through pH-controlled exonuclease (Exo)-assisted target cycle signal amplification. The AIE-labeled DNA detection probes are conjugated on Exo III encapsulated polymeric nanoparticles (NPs) via consecutive adenines (polyA). The amplified sensing functions are off during the cytoplasm delivery process, and it can be spatially switched from off to on when in the alkaline mitochondria (about pH 8) after triphenylphosphonium (TPP)-mediated mitochondrial targeting. Where the NPs degraded to release Exo III and cancer-specific mitomiRs hybridize with AIE-labeled DNA detection probes to expose the cleavage site of released Exo III, enabling spatially restricted mitomiRs imaging. The mitomiRs expression fluctuation was also realized. This study contributes to a facile strategy that could easily extend to a broad application for the understanding of mitomiRs-related pathological processes.

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.

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.

A red-emitting indolium fluorescence probe for membranes - flavonoids interactions.

The red-emitting indolium derivative compound (E)-2-(4-(diphenylamino)styryl)-1,3,3-trimethyl-3H-indol-1-ium iodide (H3) was demonstrated as a sensitive membrane fluorescence probe. The probe located at the interface of liposomes when mixed showed much fluorescence enhancement by inhibiting the twisted intramolecular charge transfer state. After ultrasonic treatment, it penetrated into lipid bilayers with the emissions leveling off and a rather large encapsulation efficiency (71.4%) in liposomes. The ζ-potential and particle size measurement confirmed that the charged indolium group was embedded deeply into lipid bilayers. The probe was then used to monitor the affinities of antioxidant flavonoids for membranes. It was verified that quercetin easily interacted with liposomes and dissociated the probe from the internal lipid within 60 s under the condition of simply mixing. The assessment of binding affinities of six flavonoids and the coincident results with their antioxidation activities indicated that it was a promising membrane probe for the study of drug bio-affinities.

A Boron 2-(2'-pyridyl) Imidazole Fluorescence Probe for Bovine Serum Albumin: Discrimination over Other Proteins and Identification of Its Denaturation.

Boron 2-(2'-pyridyl) imidazole complex (BOPIM) derivatives, BOPIM-1 and BOPIM-2, involving twisted intra-molecular charge transfer (TICT) characteristic, were investigated to probe bovine serum albumin (BSA). BOPIM-1 was demonstrated to be an efficient fluorescence probe to discriminate BSA from other proteins including human serum albumin (HSA). The encapsulation of BOPIM-1 in the hydrophobic subdomain IIA of BSA inhibited the TICT state and resulted in 70-fold emission enhancement of BOPIM-1. Besides, it was found that BOPIM-1 could distinguish denatured BSA from the native, due to the difference of 15 nm emission wavelength at least. The denaturation of BSA leads to the exposure of binding sites and resulted in an increase in the accessibility of the fluorophore to the BSA cavity, which made a great contribution to the further restraint of TICT state of BOPIM-1. The tracking of BSA denaturizing process was easily achieved, through the change of BOPIM-1 emission wavelength from 485 to 465 nm. Thus, a promising pharmacological dual-functional probe was provided for both the selective recognization of BSA and the identification of denatured BSA from the native.

OFF/ON Red-Emitting Fluorescent Probes for Casein Recognition and Quantification Based on Indolium Derivatives.

Five derivatives of 2, 3, 3-trimethyl-3H-indolium containing different electron donor groups (H1 - H5) were synthesized for the determination of proteins. H3, a sensitive red-emitting fluorescent probe, was found for the discrimination of hydrophobic proteins from hydrophilic. The OFF - ON fluorescence switch of H3 was caused by the formation of twisted intramolecular charge-transfer (TICT) state when it combined with hydrophobic proteins in aqueous buffer. There was a good linear relationship between the emission intensity of H3 and the casein concentration (r = 0.9989). Based on this, a novel casein quantitative assay method was developed, and the method was applied to determinate casein in milk powder samples. Successfully, the results were in good agreement with Biuret method. In addition, a simple and sensitive method was established to differentiate and quantify three casein components (α-, ß-, and κ-casein) due to their much different binding constants to H3 probe.