Single-Cell Diagnosis of Cancer Drug Resistance through the Differential Endocytosis of Nanoparticles between Drug-Resistant and Drug-Sensitive Cancer Cells.

Single-cell diagnosis of cancer drug resistance is highly relevant for cancer treatment, as it can be used to identify the subpopulations of drug-resistant cancer cells, reveal the sensitivity of cancer cells to treatment, and monitor the progress of cancer drug resistance. However, simple and effective methods for cancer drug resistance detection at the single-cell level are still lacking in laboratory and clinical studies. Inspired by the fact that nanoparticles with diverse physicochemical properties would generate distinct and specific interactions with drug-resistant and drug-sensitive cancer cells, which have distinctive molecular signatures, here, we have synthesized a library of fluorescent nanoparticles with various sizes, surface charges, and compositions (SiO2 nanoparticles (SNPs), organic PS-co-PAA nanoparticles (ONPs), and ZIF-8 nanoparticles (ZNPs)), thus demonstrating that the composition has a critical influence on the interaction of nanoparticles with drug-resistant cancer cells. Furthermore, the clathrin/caveolae-independent endocytosis of ZNPs together with the P-glycoprotein-related decreased cell membrane fluidity resulted in a lower cellular accumulation of ZNPs in drug-resistant cancer cells, consequently causing the distinct cellular accumulation of ZNPs between the drug-resistant and drug-sensitive cancer cells. This difference was further quantified by detecting the fluorescence signals generated by the accumulation of nanoparticles at the single-cell level via flow cytometry. Our findings provide another insight into the nanoparticle-cell interactions and offer a promising platform for the diagnosis of cancer drug resistance of various cancer cells and clinical cancer samples at the single-cell level.

Multidimensional Quantitative Measurement of Cancer Chemoresistance through Differential ZIF-8 Nanoparticle Cellular Retention.

Chemoresistance of cancer cells is conventionally quantified by half-maximal inhibitory concentration (IC50) or multidrug resistance gene 1 (MDR1) values, but these metrics can only reflect the overall drug resistance level of a cancer cell line. Meanwhile, the multidimensional evaluation of both the heterogeneity in a cell line and the drug resistance degree of each cell still presents a daunting challenge. We report here that the cellular heterogeneity, cellular cross contamination, and the proportion of chemoresistant cancer cells can be visualized via flow cytometry through the differential cellular retention of fluorescent ZIF-8 nanoparticles. In addition, we show that the degree of drug resistance exhibited by each cell subpopulation can be quantified by differing fluorescence of the drug-resistant and drug-sensitive cells in the corresponding flow cytometry profile, and the quantified metric S is highly consistent with the MDR1 expression results. Importantly, this novel strategy is applicable to various cancer cell lines, thus demonstrating a universal diagnosis platform for multidimensional, quantitative, and highly efficient diagnosis of cancer chemoresistance.

Investigations of 2D PtS2's Saturable Absorption Characteristic and Its Optimization to OPO's Operation.

A 6.2 nm-thickness platinum disulfide (PtS2) film was prepared by electron beam evaporation with post vulcanization. The nonlinear transmittance was measured by power scanning method and the modulation depth is fitted to be 13%. Based on the transmittance curve, saturable absorption parameters of PtS2 are calculated with inhomogeneously broadening mechanism, including 6.4298 × 10-19 cm-2 ground-state absorption cross-section, 2.5927 × 10-19 cm-2 excited-state absorption cross-section, and 1.043 ms excited-state lifetime. The PtS2 film combined with active time management was implemented to modulate the fundamental light of optical parametric oscillator (OPO). Owing to the nonlinear absorption property of PtS2, the operation of Q-switched OPO was optimized in both the experiment and dynamical theory. In particular, the conversion efficiency was experimentally improved by 13.2%. The pump-to-signal conversion efficiency went up to 3.29%, which is the highest conversion value reported so far. The theoretical values fit the experiment well, which are from the Gaussian rate equations with PtS2's saturable-absorption characteristic.

Monodisperse ZIF-8@dextran nanoparticles co-loaded with hydrophilic and hydrophobic functional cargos for combined near-infrared fluorescence imaging and photothermal therapy.

Impressive developments have been achieved with the use of zeolitic imidazolate framework-8 (ZIF-8) as nanocarriers for tumor theranostics in recent decades by incorporating imaging agents and therapeutic drugs within ZIF-8. However, the simultaneous immobilization of hydrophilic and hydrophobic functional molecules into ZIF-8 nanoparticles in water or organic solvents still presents a daunting challenge. Herein, we developed a new synthesis/encapsulation two-in-one (denoted as one-pot) approach to synthesize uniform dextran-modified Cy5.5&ICG@ZIF-8-Dex nanoparticles in DMSO/H2O solvent mixtures, which enabled the simultaneous encapsulation of hydrophilic indocyanine green (ICG) and hydrophobic cyanine-5.5 (Cy5.5) during the same step. It was confirmed that the one-pot approach in this mixed solvents facilitated the loading of ICG and Cy5.5 molecules. Moreover, the encapsulation of Cy5.5 and ICG within ZIF-8 nanoparticles endowed them with fluorescence imaging capability and photothermal conversion capacity, respectively. The in vivo near-infrared (NIR) fluorescent images of A549-bearing mice injected with Cy5.5&ICG@ZIF-8-Dex demonstrated sufficient accumulations of Cy5.5 at tumor sites due to the enhanced permeability and retention effect. Most impressively, the fluorescent intensity of Cy5.5&ICG@ZIF-8-Dex at tumor site was approximately 40-fold higher than that of free Cy5.5. Additionally, the results of in vivo infrared imaging and photothermal therapy of Cy5.5&ICG@ZIF-8-Dex showed enhanced therapeutic efficiency in comparison with free ICG, further confirming its tumor-targeting capability and photothermal capacity. Therefore, this multifunctional system based on ZIF-8 nanocarriers offered a potential nanoplatform for tumor-targeting theranostics, thus broadening the synthesis and applications of ZIF-8 composite nanoparticles for NIR fluorescence imaging and photothermal therapy in the biomedical field. STATEMENT OF SIGNIFICANCE: Simultaneous immobilization of hydrophilic and hydrophobic molecules into ZIF-8 nanoparticles still remains a daunting challenge. Therefore, we have developed a new synthesis/encapsulation two-in-one approach to synthesize uniform Cy5.5&ICG@ZIF-8-Dex composite nanoparticles in DMSO/H2O solvent mixtures, which enabled the simultaneous encapsulation of hydrophilic indocyanine green (ICG) and hydrophobic cyanine-5.5 (Cy5.5) functional molecules during a single step. The results showed that the co-loading of Cy5.5 and ICG within the ZIF-8 nanoparticles endowed them with a remarkable fluorescence imaging capability and photothermal conversion capacity. Based on their enhanced convenience and efficacy to simultaneously encapsulate hydrophilic and hydrophobic molecules, the multifunctional nanocarriers that were prepared in the DMSO/H2O mixed solvents provide a potential nanoplatform toward fluorescence imaging and photothermal therapy for tumor theranostics.

Mixed Solvent Method for Improving the Size Uniformity and Cargo-Loading Efficiency of ZIF-8 Nanoparticles.

Zeolitic imidazolate framework-8 (ZIF-8) nanoparticles with tunable diameters and a uniform morphology were constructed in dimethyl sulfoxide (DMSO)/H2O mixed solvents and were further decorated with dextran to improve their stability and biocompatibility. A series of reaction conditions, including the DMSO content in mixed solvents, molar ratio between precursors, growth time, and decoration of dextran, were systematically investigated. Most importantly, it was the union of DMSO and water that achieved the combined merits of both solvothermal and hydrothermal methods, namely, high uniformity and high efficiency, respectively. In addition, numerous properties of these ZIF-8 nanoparticles were subsequently studied, such as the crystal structure, surface properties, and porosity. Furthermore, composite ZIF-8 nanoparticles encapsulating various functional molecules were also successfully prepared in the same DMSO/H2O mixed solvents, thus laying the foundation for their application as nanocarriers in the biomedical field.

Lectin-Glycan-Mediated Nanoparticle Docking as a Step toward Programmable Membrane Catalysis and Adhesion in Synthetic Protocells.

The spontaneous assembly of nanoscale building blocks into continuous semipermeable membranes is a key requirement for the structuration of synthetic protocells. Engineering the functionality and programmability of these building units provides a step toward more complex cell-like entities with adaptive membrane properties. Inspired by the central role of protein (lectin)-carbohydrate interactions in cellular recognition and adhesion, we fabricate semipermeable polysaccharide-polymer microcapsules (polysaccharidosomes) with intrinsic lectin-binding properties. We employ amphiphilic polysaccharide-polymer membrane building blocks endowed with intrinsic bio-orthogonal lectin-glycan recognition sites to facilitate the reversible noncovalent docking of functionalized polymer or zeolitic nanoparticles on the polysaccharidosomes. We show that the programmed attachment of enzyme-loaded nanoparticles gives rise to a membrane-gated spatially localized cascade reaction within the protocells due to the thermoresponsiveness of the polysaccharidosome membrane, and we demonstrate that extended closely packed networks are produced via reversible lectin-mediated adhesion between the protocells. Our results provide a step toward nanoscale engineering of bioinspired cell-like materials and could have longer-term applications in synthetic virology, protobiology, and microbiosensor and microbioreactor technologies.

Near-Infrared Fluorescent and Magnetic Resonance Dual-Imaging Coacervate Nanoprobes for Trypsin Mapping and Targeted Payload Delivery of Malignant Tumors.

Trypsin-responsive near-infrared fluorescent (NIRF) and magnetic resonance (MR) dual-imaging composite nanoparticle/polypeptide coacervate nanoprobes with tunable sizes, have been constructed herein via electrostatic interaction-induced self-assembly. Considering the requirements of in vivo metabolism on nanoparticle size, three coacervate nanoprobes with diameters of around 100, 200, and 300 nm were fabricated with a polydispersity of around 0.2. These coacervate nanoprobes consist of Fe3O4 magnetic nanoparticles surface-decorated with poly acrylic acid and Cy5.5-modified poly-l-lysine (PLL-g-Cy5.5) serving as MR imaging and trypsin-responsive substrate/NIRF agents, respectively. The notable fluorescence signal from PLL-g-Cy5.5 is self-quenched due to the short distances between the fluorescent Cy5.5 molecules after construction of the coacervate nanoprobes. Remarkably, coacervate nanoprobes with a diameter of around 100 nm are selectively disintegrated into fragmented segments upon the hydrolysis of PLL by trypsin, resulting in an 18-fold amplification of the NIRF intensity in comparison with the self-assembled coacervate nanoprobes in the quenched state. Moreover, the MR imaging enhancement is also related to the disintegration of the coacervate nanoprobes. Cellular experiments and in vivo studies demonstrate that the coacervate nanoprobes exhibit remarkable trypsin-sensitive NIRF and MR dual-imaging capabilities and thus have excellent potential to serve as dual-imaging nanoprobes for the efficient mapping of malignant tumors in which trypsin is often overexpressed. In consideration of their excellent capability to enrich charged molecules, the coacervate nanoprobes provide a conceptually novel and promising platform toward in vivo trypsin mapping and controlled delivery of targeted payloads.

Structural regulation of self-assembled iron oxide/polymer microbubbles towards performance-tunable magnetic resonance/ultrasonic dual imaging agents.

Fe3O4/polymer hybrid microcapsules were prepared via a template-free route which is based on polyamine-salt aggregates (PSAs) self-assembly approach. The measurements of transmission electron microscopy (TEM) indicated that the diameter and shell thickness of the microcapsules could be tuned by varying the experimental conditions, such as the concentration of reactants and evolution time employed during the PSA assembly. The results of vibrating sample magnetometer (VSM) demonstrated that the magnetic nanoparticles content of the synthesized microcapsules was tunable and all samples exhibited superparamagnetic behavior. After filling appropriate perfluorocarbon into the inner cavities of the microcapsules, the biomedical applications of the resultant microbubbles, including ultrasonic imaging (USI) and magnetic resonance imaging (MRI), were studied in vitro. It showed that the synthesized magnetic microbubbles possessed both strong ultrasound contrast enhancement capability and high relaxation rate. The excellent acoustic and magnetic properties of these self-assembled microbubbles ensure that the Fe3O4/polymer hybrid microbubbles have great potential as MRI/USI dual-modality contrast agents.

Self-assembled microbubbles as contrast agents for ultrasound/magnetic resonance dual-modality imaging.

In this work, superparamagnetic self-assembled microbubbles (SAMBs) consisting of "Poly(acrylic acid)-Iron oxide nanoparticles-Polyamine" sandwich-like shells and tetradecafluorohexane cores were fabricated by a template-free self-assembly approach. The SAMBs exhibit not only magnetic resonance (MR) T2 imaging functionality, but also ultrasound (US) image contrast, showing great potential as US/MR dual contrast agents. The diameters of the SAMBs can be tuned easily from 450nm to 1300nm by changing the precursor ratio, and this size variation directly affects their in vitro MRI and US signals. The SAMBs also exhibit in vivo contrast enhancement capabilities in rat liver with injection through portal vein, for both MR and US imaging. Additionally, the biodistribution of SAMBs over time suggests normal systemic metabolic activity through the spleen. The results show that the Fe content in rat liver reduces to a level of which Fe cannot be detected in 45days. The SAMBs exhibit no obvious damage to the primary organs of rat during the metabolic process, indicating their good biocompatibility in vivo.

Self-assembled Fe3O4/polymer hybrid microbubble with MRI/ultrasound dual-imaging enhancement.

An Fe3O4 nanoparticle/polymer hybrid microbubble was developed using a facile self-assembly approach. This approach involves two steps, including the initial fabrication of the iron oxide nanoparticle (IONP)/polymer hybrid microcapsules via self-assembly and a subsequent gas-filling process to yield the final microbubbles. Both in vitro and in vivo experiments demonstrated that the composite gas-filled microbubbles exhibit excellent T2-weighted magnetic resonance imaging (MRI) enhancement as well as ultrasound (US) imaging enhancement capabilities. Besides, this flexible approach allows the facile control of the microbubbles' size and thus the imaging capabilities of the microbubbles through the tuning of the molar ratio between the precursors.

Efficient preparation of magnetic quantum dot barcodes.

Quantum dot (QD)-encoded magnetic barcodes were prepared through a highly efficient membrane emulsification-solvent evaporation approach. Our study demonstrates the great potential of these QD-encoded magnetic barcodes for both magnetic separation and multiplex suspension assays.