Ultrafast Subcellular Biolabeling and Bioresponsive Real-Time Monitoring for Targeting Cancer Theranostics.

Cell heterogeneity poses a formidable challenge for tumor theranostics, requiring high-resolution strategies for intercellular bioanalysis between single cells. Nanoelectrode-based electrochemical analysis has attracted much attention due to its advantages of label-free characteristics, relatively low cost, and ultra-high resolution for single-cell analysis. Here, we have designed and developed a subcellular biolabeling and bioresponsive real-time monitoring strategy for precise bioimaging-guided cancer diagnosis and theranostics. Our observations revealed the apparent intracellular migration of biosynthetic Au nanoclusters (Au NCs) at different subcellular locations, i.e., from the mitochondria to the mitochondrion-free region in the cytoplasm, which may be helpful for controlling over the biosynthesis of Au NCs. Considering the precise biolabeling advantage of the intracellular biosynthetic Au NCs for biomedical imaging of cancers, it is important to realize the biosynthetic Au NC-enabled precise control in real-time theranostics of cancer cells. Therefore, this work raises the possibility to achieve subcellular monitoring of H2O2 for targeting cancer theranostics, thereby providing a new way to explore the underlying mechanism and imaging-guided tumor theranostics.

Ultraprecise Real-Time Monitoring of Single Cells in Tumors in Response to Metal Ion-Mediated RNA Delivery.

With the deepening of cancer clinical research, miRNAs provide new ideas for molecular diagnosis and treatment of tumors. Improving the molecular delivery efficiency of miRNA is the key to the success of miRNA therapy. We have established self-assembly diagnosis and treatment technologies that can be used to achieve accurate targeting and "cargo" delivery at the cellular level. This technology builds a miRNA (let-7a) delivery system based on metal precursor [Au(III) and Fe(II)]-mediated tumor microenvironmental response to realize the self-assembly of Au&Fe-miRNA complexes for precise real-time imaging of tumor cells and targeted therapy. To accurately measure the changes in reactive oxygen species during complex formation in real time at the single-cell level, we employed small-size nanoscale devices as analytical tools. This study proposes an electrochemical sensor based on carbon fiber electrodes for ultraprecise and multiple monitoring of metal-ion-mediated miRNA delivery systems, precisely realizing targeted tracking of tumors and effective intervention inhibition.

Bio responsive self-assembly of Au-miRNAs for targeted cancer theranostics.

BACKGROUND: MicroRNA (miRNA) therapeutics are a promising approach to cancer treatment. However, this method faces considerable challenges to achieve tissue-specific, efficient, and safe delivery of miRNAs in vivo. METHODS: Herein, we developed a miRNA delivery system based on the in situ self-assembly of Au-miRNA nanocomplexes (Au-miRNA NCs). Within the cancer microenvironment, we constructed in situ self-assembled Au-miRNA NCs by coincubating gold salt and tumor suppressor mimics, such as let-7a, miRNA-34a, and miRNA-200a. FINDINGS: The in vitro experiments demonstrated that characteristic in situ self-assembled Au-miRNA NCs were present in cancer cells and can be taken up to inhibit the proliferation of cancer cells effectively. Most importantly, as proven in subcutaneous tumor treatment models, Au-miRNA NCs were especially useful for accurate target imaging and tumor suppression, with significantly enhanced antitumor effects for combination therapy. INTERPRETATION: These observations highlight that a new strategy for the in situ biosynthesis of Au-let-7a NCs, Au-miR-34a NCs, and Au-miR-200a NCs is feasible, and this may assist in the delivery of more miRNA to tumor cells for cancer treatment. This work opens up new opportunities for the development of miRNA tumor therapy strategies. FUNDING: National Natural Science Foundation of China (91753106); Primary Research & Development Plan of Jiangsu Province (BE2019716); National Key Research and Development Program of China (2017YFA0205300).

Manganese oxide doped carbon dots for temperature-responsive biosensing and target bioimaging.

We report on the synthesis of manganese oxide doped CDs (MnOx-CDs) by a hydrothermal strategy using manganese (III) acetylacetonate (Mn(III) (C5H7O2)3) as the only raw materials. The MnOx-CDs exhibit water solubility, favorable biocompatibility, low cytotoxicity, and show blue fluorescence with excitation/emission maxima at 326/442 nm with a quantum yield of 11.3%, allowing efficient cellular imaging. The MnOx-CDs have a reversible temperature-sensitive fluorescent property in vitro within 10-60 °C, which can also be used as a sensitive thermometer in living cells. By a scratch assay, the MnOx-CDs can restrain the migration of HepG2 cancer cells, which make the MnOx-CDs be attractive candidates for liver cancer adjuvant treatment. Besides, the fluorescence of the MnOx-CDs is quenched in the presence of Fe3+ due to the formation of a nonfluorescent MnOx-CDs-Fe3+ complex between oxygen-containing groups on the surface of MnOx-CDs and Fe3+, and the quenched fluorescence of MnOx-CDs can be turn-on by dissociation of MnOx-CDs-Fe3+ complexes by biothiols including L-cysteine, homocysteine and glutathione. Therefore, the Fe3+ and biothiols can be sequentially detected with high reliability and accuracy via exploiting the on-off-on nanosensor at room temperature, respectively. Further application to detection biothiols in human serum indicated that the probe was practicality and feasibility in medical field.

Fluorescence light up detection of aluminium ion and imaging in live cells based on the aggregation-induced emission enhancement of thiolated gold nanoclusters.

In this paper, a new strategy was presented for fluorescence labeling and imaging Al3+ in live cells with excess aluminum ions using thiolated fluorescence gold nanoclusters (Au NCs). The glutathione (GSH)-capped Au NCs were prepared via a green, facile one-pot method in aqueous solution and displayed excellent stability, ultrasmall size, monodispersity, and larger Stokes shift, which exhibits a relatively weak fluorescence at 650 nm Al3+-induced fluorescence enhancement of the GSH-Au NCs can be observed due to Al3+-triggered aggregation-induced emission (AIE) effect, which allows the role of GSH-Au NCs as a fluorescence light-up probe for detection of Al3+. Moreover, it was demonstrated that the fluorescence probe for Al3+ showed a wide detection range from 100 to 600 µM and good selectivity against other metal ions and common biomolecule. Furthermore, due to the advantages of excellent biocompatibility, low toxicity, red emission and high specificity, the proposed GSH-Au NCs fluorescence probes are suitable for the imaging of high concentrations of aluminum ions in cells, which can be applied to the diagnosis of cellular aluminum poisoning.

Glutathione Induced Transformation of Partially Hollow Gold-Silver Nanocages for Cancer Diagnosis and Photothermal Therapy.

Gold-silver nanocages (GSNCs) are widely used in cancer imaging and therapy due to excellent biocompatibility, internal hollow structures, and tunable optical properties. However, their possible responses toward the tumor microenvironment are still not well understood. In this study, it is demonstrated that a kind of relatively small sized (35 nm) and partially hollow GSNCs (absorbance centered at 532 nm) can enhance the intrinsic photoacoustic imaging performances for blood vessels around tumor sites. More importantly, the high concentration of glutathione around the tumor cells' microenvironment may induce the aggregation, disintegration, and agglomeration of these GSNCs sequentially, allowing significant shifts in the absorbance spectrum of GSNCs to the near-infrared (NIR) region. This enhanced absorbance in the NIR region entails the significant photothermal therapy (PTT) effect. In vivo experiments, including photoacoustic microscopy (PAM) for cancer diagnosis and PTT in tumor model mice, also show coincident consequences. Taken together, the slightly hollow GSNCs may assist PAM-based tumor diagnosis and induce a tumor targeted PTT effect. This work paves a new avenue for the development of an alternative tumor diagnostic and therapeutic strategy.

Aggregation-Induced Electrochemiluminescence by Metal-Binding Protein Responsive Hydrogel Scaffolds.

Functionalized hydrogels have aroused general interest due to their versatile applications in biomaterial fields. This work reports a hydrogel network composed of gold nanoclusters linked with bivalent cations such as Ca2+ , Mg2+ , and Zn2+ . The hydrogel exhibits both aggregation-induced emission (AIE) and aggregation-induced electrochemiluminescence (AIECL) effects. Most noteworthy, the AIECL effect (≈50-fold enhancement) is even more significant than the corresponding AIE effect (approximately fivefold enhancement). Calmodulin, a Ca2+ binding protein, may efficiently regulate the AIECL dynamics after specific binding of the Ca2+ linker, with the linear range from 0.3 to 50 µg mL-1 and a limit of detection of 0.1 µg mL-1 . Considering the important roles of bivalent cations in the life system, these results may pave a new avenue for the design of a biomolecule-responsive AIECL-type hydrogel with multifunctional biomedical purposes.

Mercaptopyrimidine-Conjugated Gold Nanoclusters as Nanoantibiotics for Combating Multidrug-Resistant Superbugs.

Widespread bacterial resistance induced by the abuse of antibiotics eagerly needs the exploitation of novel antimicrobial agents and strategies. Gold nanoclusters (Au NCs) have recently emerged as an innovative nanomedicine, but study on their antibacterial properties especially toward multidrug resistant (MDR) bacteria is scarce. Herein, we demonstrate that a novel class of Au NCs, mercaptopyrimidine conjugated Au NCs, can act as potent nanoantibiotics targeting these intractable superbugs in vitro and in vivo, without induction of bacterial antibiotic resistance and noticeable cytotoxicity to mammalian cells. The Au NCs kill these superbugs through a combined mechanism including cell membrane destruction, DNA damage, and reactive oxygen species (ROS) generation, and exhibit excellent treatment effects in both macrophages and animal infection models induced by methicillin-resistant Staphylococcus aureus as representative. Moreover, the induction of intracellular ROS production in bacterial cells mainly attributed to the Au NCs' intrinsic oxidase- and peroxidase-like catalytic activities has been demonstrated for the first time.