Molecular Nanoprobes Research Articles

A Smart Intracellular Self-Assembling Bioorthogonal Raman Active Nanoprobe for Targeted Tumor Imaging.

Inspired by the principle of in situ self-assembly, the development of enzyme-activated molecular nanoprobes can have a profound impact on targeted tumor detection. However, despite their intrinsic promise, obtaining an optical readout of enzyme activity with high specificity in native milieu has proven to be challenging. Here, a fundamentally new class of Raman-active self-assembling bioorthogonal enzyme recognition (nanoSABER) probes for targeted tumor imaging is reported. This class of Raman probes presents narrow spectral bands reflecting their vibrational fingerprints and offers an attractive solution for optical imaging at different bio-organization levels. The optical beacon harnesses an enzyme-responsive peptide sequence, unique tumor-penetrating properties, and vibrational tags with stretching frequencies in the cell-silent Raman window. The design of nanoSABER is tailored and engineered to transform into a supramolecular structure exhibiting distinct vibrational signatures in presence of target enzyme, creating a direct causality between enzyme activity and Raman signal. Through the integration of substrate-specific for tumor-associated enzyme legumain, unique capabilities of nanoSABER for imaging enzyme activity at molecular, cellular, and tissue levels in combination with machine learning models are shown. These results demonstrate that the nanoSABER probe may serve as a versatile platform for Raman-based recognition of tumor aggressiveness, drug accumulation, and therapeutic response.

In Situ Noninvasive Observation of Nitric Oxide Fluctuation in SARS-CoV-2 Infection In Vivo by Organic Near-Infrared-II Fluorescent Molecular Nanoprobes.

The pathogenesis understanding of SARS-CoV-2 infection is crucial to prevent the rampant spread of COVID-19 and its contribution to deterioration in health, even death. Nitric oxide (NO), a crucial molecule involved in signal transduction and cytotoxicity, is a possible key regulator in the occurrence and development of COVID-19. However, understanding the pathogenesis of NO in SARS-CoV-2 infection is still in its infancy due to the lack of suitable in situ monitoring probes of NO fluctuation in the complex SARS-CoV-2 infection environment in deep lung tissues. Herein, we developed an activatable near-infrared-II fluorescent molecular nanoprobe (OSNP) that uncages high-resolution and deep-tissue-penetrating near-infrared-II fluorescence signal in specific response to NO for in situ and noninvasive visualization of NO fluctuation in a SARS-CoV-2 infection mouse model in lung tissues. In vivo visualization revealed that the NO level is a positive relationship with SARS-CoV-2 infection progress. With the assistance of immuno-histochemical analyses, we uncovered the NO-involved pathological mechanism, that being the improved NO level is associated with an increase in inducible NO synthase rather than endothelial NO synthase. Our study not only provides the example of a near-infrared-II fluorescent imaging of NO in SARS-CoV-2 infection but also provides opportunities to uncover tunderlying pathomechanism of NO for SARS-Cov-2 infections.

Dynamics of molecular rotors in bulk superfluid helium.

Molecules immersed in liquid helium are excellent probes of superfluidity. Their electronic, vibrational, and rotational dynamics provide valuable clues about the superfluid at the nanoscale. Here we report on the experimental study of the laser-induced rotation of helium dimers inside the superfluid 4He bath at variable temperature. The coherent rotational dynamics of [Formula: see text] is initiated in a controlled way by ultrashort laser pulses and tracked by means of time-resolved laser-induced fluorescence. We detect the decay of rotational coherence on the nanosecond time scale and investigate the effects of temperature on the decoherence rate. The observed temperature dependence suggests a nonequilibrium evolution of the quantum bath, accompanied by the emission of the wave of second sound. The method offers ways of studying superfluidity with molecular nanoprobes under variable thermodynamic conditions.

Water-Soluble Chemically Precise Fluorinated Molecular Clusters for Interference-Free Multiplex 19F MRI in Living Mice.

Fluorine-19 magnetic resonance imaging (19F MRI) is gaining widespread interest from the fields of biomolecule detection, cell tracking, and diagnosis, benefiting from its negligible background, deep tissue penetration, and multispectral capacity. However, a wide range of 19F MRI probes are in great demand for the development of multispectral 19F MRI due to the limited number of high-performance 19F MRI probes. Herein, we report a type of water-soluble molecular 19F MRI nanoprobe by conjugating fluorine-containing moieties with a polyhedral oligomeric silsesquioxane (POSS) cluster for multispectral color-coded 19F MRI. These chemically precise fluorinated molecular clusters are of excellent aqueous solubility with relatively high 19F contents and of single 19F resonance frequency with suitable longitudinal and transverse relaxation times for high-performance 19F MRI. We construct three POSS-based molecular nanoprobes with distinct 19F chemical shifts at -71.91, -123.23, and -60.18 ppm and achieve interference-free multispectral color-coded 19F MRI of labeled cells in vitro and in vivo. Moreover, in vivo 19F MRI reveals that these molecular nanoprobes could selectively accumulate in tumors and undergo rapid renal clearance afterward, illustrating their favorable in vivo behavior for biomedical applications. This study provides an efficient strategy to expand the 19F probe libraries for multispectral 19F MRI in biomedical research.

Image‐guided diagnosis and treatment of glioblastoma

AbstractGlioblastoma (GBM) is the most aggressive primary brain tumor with poor prognosis and high recurrence rate. The presence of the blood–brain barrier (BBB) prevents diagnostic and therapeutic drugs from penetrating and working in GBM. In traditional surgical treatment, it is difficult to completely distinguish the boundary between tumor and surrounding normal tissue, resulting in incomplete resection of tumor. Therefore, the diagnosis and treatment of GBM are very challenging. Several molecular probes and nanoprobes have been reported to successfully penetrate the BBB, selectively target and accumulate in GBM to achieve in situ imaging of brain tumors, thus achieving accurate diagnosis and treatment of orthotopic or non‐orthotopic GBM. This paper reviews the advances of molecular probes and nanoprobes in image‐guided diagnosis and treatment of GBM. The design principle, application, and advantages of each probe are enumerated in detail. Finally, the prospects and potential challenges of probes in the diagnosis and treatment of GBM are discussed with a view to further promote the study and application of novel imaging probes in GBM.

Engineering molecular nanoprobes to target early atherosclerosis: Precise diagnostic tools and promising therapeutic carriers.

Atherosclerosis, an inflammation-driven chronic blood vessel disease, is a major contributor to devastating cardiovascular events, bringing serious social and economic burdens. Currently, non-invasive diagnostic and therapeutic techniques in combination with novel nanosized materials as well as established molecular targets are under active investigation to develop integrated molecular imaging approaches, precisely visualizing and/or even effectively reversing early-stage plaques. Besides, mechanistic investigation in the past decades provides many potent candidates extensively involved in the initiation and progression of atherosclerosis. Recent hotly-studied imaging nanoprobes for detecting early plaques mainly including optical nanoprobes, photoacoustic nanoprobes, magnetic resonance nanoprobes, positron emission tomography nanoprobes, and other dual- and multi-modality imaging nanoprobes, have been proven to be surface functionalized with important molecular targets, which occupy tailored physical and biological properties for atherogenesis. Of note, these engineering nanoprobes provide long blood-pool residence and specific molecular targeting, which allows efficient recognition of early-stage atherosclerotic plaques and thereby function as a novel type of precise diagnostic tools as well as potential therapeutic carriers of anti-atherosclerosis drugs. There have been no available nanoprobes applied in the clinics so far, although many newly emerged nanoprobes, as exemplified by aggregation-induced emission nanoprobes and TiO2 nanoprobes, have been tested for cell lines in vitro and atherogenic animal models in vivo, achieving good experimental effects. Therefore, there is an urgent call to translate these preclinical results for nanoprobes into clinical trials. For this reason, this review aims to give an overview of currently investigated nanoprobes in the context of atherosclerosis, summarize relevant published studies showing applications of different kinds of formulated nanoprobes in early detection and reverse of plaques, discuss recent advances and some limitations thereof, and provide some insights into the development of the new generation of more precise and efficient molecular nanoprobes, with a critical property of specifically targeting early atherosclerosis.

Multimodal ultra-small CoFe-WOx nanohybrids synthesized by a pilot microfluidic system

Among all the nanomaterials, ultra-small nanohybrids have gained considerable attention as drugs, contrast agents, molecular nanoprobes, and catalysts owing to their multi-functionality and size effect. However, their extension and commercialization are restricted due to low synthetic efficiency and uncontrollable structure. Herein, a pilot microfluidic system (PMS) was built to successfully synthesize the ultra-small (2.3 ± 0.7 nm) CoFe-WOx nanohybrids (NHs) with a tiny CoFe core and a mixed metal oxides shell (CoWO4-Fe2WO6-WO3). Furthermore, the CoFe-WOx NHs displayed excellent multi-modal imaging properties, which could be observed optically via a dark-field microscope, imaged through X-ray computed tomography and magnetic resonance imaging device. For the first time, hybridization between metal cores and metal-oxide shells produced two strong absorption peaks in the long band of near infrared radiation, one at 1450 nm and the other at 1980 nm. Overall, CoFe-WOx NHs emonstrated distinct magnetic and optical properties and multimode imaging capabilities due to their microstructure and composition. These findings may prove to be effective in finding alternatives for nanoprobes that can be used for visible/infrared/MRI/CT imaging, as well as pre-drugs for photothermal/photodynamic and magnetothermal/magneto-dynamic therapies.

Multifunctional management of gold nanoparticles for improved applications

Increasingly, nanotechnology has the potential to be applied in disease detection or treatment. Nanotechnology advancements have resulted in novel and improved biomedical nanomaterials. Liposomes, polymeric micelles, graphene, carbon nanotubes, quantum dots, ferro ferric oxide nanoparticles, gold nanoparticles (Au NPs), and so on are some of the most commonly used nanomaterials in biomedical applications. Because of its unique optical, electrical, sensing and biological capabilities, Au NPs have been regarded as the most fascinating nanomaterial. Medical imaging, drug administration and tumour therapy are all possible applications of Au NPs in the early identification and diagnosis and treatment of illnesses. This review focuses on some recent breakthroughs in the management of Au NPs as drug carriers for the intracellular delivery of therapies and as molecular nanoprobes for the detection and monitoring of target molecules.

Biosensors for Caspase-3: From chemical methodologies to biomedical applications

Caspase-3 plays irreplaceable roles in apoptosis and related diseases. An imbalance in the measured levels of Caspase-3 is implicated in irreversible apoptosis. Therefore, the detection of Caspase-3 is of great significance for apoptosis imaging and the evaluation effect of early tumor treatment and other diseases. Herein, advances in the recent innovations of Caspase-3 response fluorescence biosensors, including molecular probes and nanoprobes, are systematically summarized in sections corresponding. The performances of various luminescence probes in Caspase-3 detection are discussed intensively in the design strategy of chemical structure, response mechanism and biological application. Finally, the current challenges and prospects of the design of new Caspase-3 responsive fluorescence probes for apoptosis imaging, or similar molecular event are proposed.

Application of Molecular Nanoprobes in the Analysis of Differentially Expressed Genes and Prognostic Models of Primary Hepatocellular Carcinoma.

Analyzing hub genes related to tumorigenesis based on biological big data has recently become a hotspot in biomedicine. Nanoprobes, nanobodies and theranostic molecules targeting hub genes delivered by nanocarriers have been widely applied in tumor theranostics. Hepatocellular carcinoma (HCC) is one of the most common cancers, with a poor prognosis and high mortality. Identifying hub genes according to the gene expression levels and constructing prognostic signatures related to the onset and outcome of HCC will be of great significance. In this study, the expression profiles of HCC and normal tissue were obtained from the GEO database and analyzed by GEO₂R to identify DEGs. GO terms and KEGG pathways were enriched in DAVID software. The STRING database was consulted to find protein-protein interactions between proteins encoded by the DEGs, which were visualized by Cytoscape. Then, overall survival associated with the hub genes was calculated by the Kaplan-Meier plotter online tool, and verification of the results was carried out on TCGA samples and their corresponding clinical information. A total of 603 DEGs were obtained, of which 479 were upregulated and 124 were downregulated. PPI networks including 603 DEGs and 18 clusters were constructed, of which 7 clusters with MCODE score ≥3 and nodes ≥5 were selected. The 5 genes with the highest degrees of connectivity were identified as hub genes, and a prognostic model was constructed. The expression and prognostic potential of this model was validated on TCGA clinical data. In conclusion, a five-gene signature (TOP2A, PCNA, AURKA, CDC20, CCNB2) overexpressed inHCC was identified, and a prognostic model was constructed. This gene signature may act as a prognostic model for HCC and provide potential targets of nanotechnology.

Molecular Targeting Nanoprobes with Non-Overlap Emission in the Second Near-Infrared Window for in Vivo Two-Color Colocalization of Immune Cells.

Monitoring specific immune cells in vivo will provide significant information for improving the therapeutic effect of immunotherapy. Herein, the in vivo two-color fluorescence molecular imaging of an important immune cell, myeloid-derived suppressor cell (MDSC), was realized by using quantum dot (QD)-based nanoprobes with non-overlap emission in the second near-infrared window (NIR-II, 1000-1700 nm). NIR-IIa and NIR-IIb QDs were conjugated with two MDSC-specific antibodies, respectively, and targeted the in vivo MDSCs together. Due to the suppressed photon scattering and diminished autofluorescence in the NIR-II window, the distribution of MDSCs in different organs and tissues was clearly revealed in a non-invasive way by the colocalization of two-color fluorescence from nanoprobes. The high-resolution imaging further confirmed the exact distribution of MDSCs in tumor immune microenvironment (TIME). Our results demonstrated that NIR-II fluorescence nanoprobes with molecular targeting ability provided a powerful tool for monitoring the dynamic change of immune cell populations in TIME in vivo, thus guiding the choice of clinical medicine and evaluating the therapeutic effect.

Recent Advances in Fluorescence Imaging of Bioactive Molecules in Neurons and in Vivo

Brain possesses a unique chemical composition and reactivity. Bioactive molecules in brains including metal ions, reactive oxygen species, neurotransmitters and neurotransmitter hydrolases play multifarious and pivotal roles in the function of brain. These molecules keep steady-state concentrations for maintaining the brain's structure and function. However, the abnormal changes in bioactive molecular levels are associated with various brain diseases such as Parkinson's disease, Alzheimer's disease, depression, and so on. Therefore, to explore the concentration and regulation of those molecules in brains is particularly essential for revealing the mysteries of brains and understanding the occurrence and development of brain diseases. Fluorescence imaging is a powerful tool to track bioactive molecules in neurons and in brain because of its remarkable advantages, such as high temporal-spatial resolution, excellent biocompatibility and high sensitivity. In recent years, with the development of fluorescence imaging technology, a variety of fluorescent probes for brain and neurons imaging have been reported. In this review, we summarize the progress of molecular probes, nanoprobes and protein probes for the detection of bioactive molecules in neurons and in brain. Furthermore, future research for fluorescence imaging are prospected.

Plasmonic Nanoprobes for in Vivo Multimodal Sensing and Bioimaging of MicroRNA within Plants.

Monitoring gene expression within whole plants is critical for many applications ranging from plant biology to agricultural biotechnology and biofuel development; however, no method currently exists for in vivo monitoring of genomic targets in plant systems without requiring sample extraction. Herein, we report a unique multimodal method based on plasmonic nanoprobes capable of in vivo imaging and biosensing of microRNA biotargets within whole plant leaves by integrating three different and complementary techniques: surface-enhanced Raman scattering (SERS), X-ray fluorescence (XRF), and plasmonics-enhanced two-photon luminescence (TPL). The method developed uses plasmonic nanostars, which not only provide large Raman signal enhancement but also allow for localization and quantification by XRF and plasmonics-enhanced TPL, owing to gold content and high two-photon luminescence cross sections. Our method uses inverse molecular sentinel nanoprobes for SERS bioimaging of microRNA within Arabidopsis thaliana leaves to provide a dynamic SERS map of detected microRNA targets while also quantifying nanoprobe concentrations using XRF and TPL. The nanoprobes were observed to occupy the intercellular spaces upon infiltration into the leaf tissues. This report lays the foundation for the use of plasmonic nanoprobes for in vivo functional imaging of nucleic acid biotargets in whole plants, a tool that will revolutionize bioengineering research by allowing the study of these biotargets with previously unmet spatial and temporal resolution, 200 μm and 30 min, respectively.

Eu-Doped Ceria Nanocrystals as Nanoenzyme Fluorescent Probes for Biosensing

Molecular nanoprobes with intrinsic enzyme-like activity represent a new wave of technology for rapid and sensitive detection of molecular targets. This work reports synthesis and characterization ...

DNA assay based on Nanoceria as Fluorescence Quenchers (NanoCeracQ DNA assay)

Functional nanomaterials with fluorescent or quenching abilities are important for the development of molecular probes for detection and studies of nucleic acids. Here, we describe a new class of molecular nanoprobes, the NanoCeracQ that uses nanoceria particles as a nanoquencher of fluorescent oligonucleotides for rapid and sensitive detection of DNA sequences and hybridization events. We show that nanoceria forms stable and reversible bionanoconjugates with oligonucleotides and can specifically recognize and detect DNA sequences in a single step. In absence of the target DNA, the nanoprobe produced minimal background fluorescence due to the high quenching efficiency of nanoceria. Competitive binding of the target induced a concentration dependent increase in the fluorescence signal due to hybridization and release of the fluorescent tag from the nanoparticle surface. The nanoprobe enabled sensitive detection of the complementary strand with a detection limit of 0.12 nM, using a single step procedure. The results show that biofunctionalized nanoceria can be used as a universal nanoquencher and nanosensing platform for fluorescent DNA detection and studies of nucleic acid interactions. This approach can find broad applications in molecular diagnostics, sensor development, gene expression profiling, imaging and forensic analysis.

Unique Roles of Gold Nanoparticles in Drug Delivery, Targeting and Imaging Applications.

Nanotechnology has become more and more potentially used in diagnosis or treatment of diseases. Advances in nanotechnology have led to new and improved nanomaterials in biomedical applications. Common nanomaterials applicable in biomedical applications include liposomes, polymeric micelles, graphene, carbon nanotubes, quantum dots, ferroferric oxide nanoparticles, gold nanoparticles (Au NPs), and so on. Among them, Au NPs have been considered as the most interesting nanomaterial because of its unique optical, electronic, sensing and biochemical properties. Au NPs have been potentially applied for medical imaging, drug delivery, and tumor therapy in the early detection, diagnosis, and treatment of diseases. This review focuses on some recent advances in the use of Au NPs as drug carriers for the intracellular delivery of therapeutics and as molecular nanoprobes for the detection and monitoring of target molecules.

Croconaine nanoparticles with enhanced tumor accumulation for multimodality cancer theranostics.

A novel nanoparticle self-assembled by polyethylene glycol (PEG) modified croconaine dye (CR780) is presented for photoacoustic (PA)/near-infrared (NIR) fluorescence imaging-guided photothermal therapy (PTT). The simple PEGylation made CR780 amphiphilic, and led to their self-assembly into well-defined and uniform nanostructures with size tunable by controlling the assembly conditions. The CR780-PEG5K not only displayed the strength of small molecules (including rapid distribution to different organs, fast renal clearance and minimal accumulation to normal tissues), but also demonstrated the advantages of nanomaterials (including high physiological stability, multimodal theranostic ability, high tumor accumulation and retention). These facilely synthesized molecular nanoprobes showed great clinical translation potential as a versatile theranostic agent.

The risk assessment of Gd2O3:Yb3+/Er3+ nanocomposites as dual-modal nanoprobes for magnetic and fluorescence imaging

Our group has synthesized Gd2O3:Yb3+/Er3+ nanocomposites as magnetic/fluorescence imaging successfully in the previous study, which exhibit good uniformity and monodispersibility with a mean size of 7.4 nm. However, their systematic risk assessment remains unknown. In this article, the in vitro biocompatibility of the Gd2O3:Yb3+/Er3+ was assessed on the basis of cell viability and apoptosis. In vivo immunotoxicity was evaluated by monitoring the product of reactive oxygen species (ROS), clusters of differentiation (CD) markers, and superoxide dismutase (SOD) in Balb/c mice. No significant differences were found in cell viability, apoptosis, and immunotoxicity between our Gd2O3:Yb3+/Er3+ and gadodiamide which are used commonly in clinical. Few nanoprobes were localized in the phagosomes of the liver, heart, lung, spleen, kidney, brain, and tumor under the transmission electron microscopy (TEM) images. In addition, our products reveal good T1-weighted contrast enhancement of xenografted murine tumor. Therefore, the above results may contribute to the effective application of Gd2O3:Yb3+/Er3+ as molecular imaging contrast agents and dual-modal nanoprobes for cancer detection.

Using ordered nanostructures on optical fiber tips as molecular nanoprobes

Using ordered nanostructures on optical fiber tips as molecular nanoprobes

Noninvasive monitoring of early antiangiogenic therapy response in human nasopharyngeal carcinoma xenograft model using MRI with RGD-conjugated ultrasmall superparamagnetic iron oxide nanoparticles.

PurposeArginine-glycine-aspartic acid (RGD)-based nanoprobes allow specific imaging of integrin αvβ3, a protein overexpressed during angiogenesis. Therefore, this study applied a novel RGD-coupled, polyacrylic acid (PAA)-coated ultrasmall superparamagnetic iron oxide (USPIO) (referred to as RGD-PAA-USPIO) in order to detect tumor angiogenesis and assess the early response to antiangiogenic treatment in human nasopharyngeal carcinoma (NPC) xenograft model by magnetic resonance imaging (MRI).Materials and methodsThe binding specificity of RGD-PAA-USPIO with human umbilical vein endothelial cells (HUVECs) was confirmed by Prussian blue staining and transmission electron microscopy in vitro. The tumor targeting of RGD-PAA-USPIO was evaluated in the NPC xenograft model. Later, mice bearing NPC underwent MRI at baseline and after 4 and 14 days of consecutive treatment with Endostar or phosphate-buffered saline (n=10 per group).ResultsThe specific uptake of the RGD-PAA-USPIO nanoparticles was mainly dependent on the interaction between RGD and integrin αvβ3 of HUVECs. The tumor targeting of RGD-PAA-USPIO was observed in the NPC xenograft model. Moreover, the T2 relaxation time of mice in the Endostar-treated group decreased significantly compared with those in the control group both on days 4 and 14, consistent with the immunofluorescence results of CD31 and CD61 (P<0.05).ConclusionThis study demonstrated that the magnetic resonance molecular nanoprobes, RGD-PAA-USPIOs, allow noninvasive in vivo imaging of tumor angiogenesis and assessment of the early response to antiangiogenic treatment in NPC xenograft model, favoring its potential clinical translation.