Size and shape control of microgel-encapsulating tumor spheroid via a user-friendly solenoid valve-based sorter and its application on precise drug testing.

The precision of previous cancer research based on tumor spheroids, especially the microgel-encapsulating tumor spheroids, was limited by the high heterogeneity in the tumor spheroid size and shape. Here, we reported a user-friendly solenoid valve-based sorter to reduce this heterogeneity. The artificial intelligence algorithm was employed to detect and segmentate the tumor spheroids in real-time for the size and shape calculation. A simple off-chip solenoid valve-based sorting actuation module was proposed to sort out target tumor spheroids with the desired size and shape. Utilizing the developed sorter, we successfully uncovered the drug response variations on cisplatin of lung tumor spheroids in the same population but with different sizes and shapes. Moreover, with this sorter, the precision of drug testing on the spheroid population level was improved to a level comparable to the precise but complex single spheroid analysis. The developed sorter also exhibits significant potential for organoid morphology and sorting for precision medicine research.

PEG-SeNPs as therapeutic agents inhibiting apoptosis and inflammation of cells infected with H1N1 influenza A virus.

The rapid variation of influenza challenges vaccines and treatments, which makes an urgent task to develop the high-efficiency and low-toxicity new anti-influenza virus drugs. Selenium is one of the essential trace elements for the human body that possesses a good antiviral activity. In this study, we assessed anti-influenza A virus (H1N1) activity of polyethylene glycol (PEG)-modified gray selenium nanoparticles (PEG-SeNPs) on Madin-Darby Canine Kidney (MDCK) cells in vitro. CCK-8 assay showed that PEG-SeNPs had a protective effect on H1N1-infected MDCK cells. Moreover, PEG-SeNPs significantly reduced the mRNA level of H1N1. TUNEL-DAPI test showed that DNA damage reached a high level but effectively prevented after PEG-SeNPs treatment. Meanwhile, JC-1, Annexin V-FITC and cell cycle assay demonstrated the apoptosis induced by H1N1 was reduced greatly when treated with PEG-SeNPs. Furthermore, the downregulation of p-ATM, p-ATR and P53 protein, along with the upregualation of AKT protein indicated that PEG-SeNPs could inhibit H1N1-induced cell apoptosis through reactive oxygen species (ROS)-mediated related signaling pathways. Finally, Cytokine detection demonstrated PEG-SeNPs inhibited the production of pro-inflammatory factors after infection, including IL-1ß, IL-5, IL-6, and TNF-α. To sum up, PEG-SeNPs might become a new potential anti-H1N1 influenza virus drug due to its antiviral and anti-inflammatory activity.

Copolythiophene-derived colorimetric and fluorometric sensor for visually supersensitive determination of lipopolysaccharide.

3-Phenylthiophene-based water-soluble copolythiophenes (CPT1) were designed for colorimetric and fluorometric detection of lipopolysaccharide (LPS). The sensor (CPT1-C) shows a high selectivity to LPS in the presence of other negatively charged bioanalytes as well an extreme sensitivity with the detection limit at picomolar level, which is the lowest ever achieved among all synthetic LPS sensors available thus far. Significantly, the sensing interaction can be apparently observed by the naked eyes, which presents a great advantage for its practical applications. The appealing performance of sensor was demonstrated to originate from the multiple electrostatic and hydrophobic cooperative interactions, synergetic with signal amplification via the conformational change of the 3-phenylthiophene-based copolymer main chain. As a straightforward application, CPT1-C is capable of rapidly discriminating the Gram-negative bacteria (with LPS in the membrane) from Gram-positive bacteria (without LPS).

N-alkyl-polyethylenimine stabilized iron oxide nanoparticles as MRI visible transfection agents.

An amphiphilic polymer, alkylated branched polyethylenimine (N-Alkyl-PEI), is synthesized and used for stabilization of hydrophobic superparamagnetic iron oxide (SPIO) nanocrystals in aqueous phase. Such composite particles are monodisperse without aggregation in physiological buffer as verified by dynamic light scattering (DLS) and transmission electron microscopy (TEM). The nanocomposite system is capable of binding and delivering plasmid DNA for gene transfection while maintaining magnetic properties and biocompatibility. Transfection of cells showed that N-Alkyl-PEI2k stabilized magnetite nanoparticles were most effective in gene transfection comparing to unmodified PEI2k and PEI25k agents. Obvious MR signal darkening of transfected cells was observed under a clinical 3T MRI scanner. This multifunctional nanocomposite system provides a safe and efficient method for gene delivery with non-invasive imaging monitoring capability.

Self-assembly of SiO2/Gd-DTPA-polyethylenimine nanocomposites as magnetic resonance imaging probes.

Controlled self-assembly of organic/inorganic magnetic hybrid materials have important applications in magnetic resonance imaging (MRI). In this study, a widely used polycation polyethylenimine was conjugated with gadopentetic acid (Gd-DTPA) as a gadolinium bearing polyelectrolyte (Gd-DTPA-PEI). Next, multilayers of Gd-DTPA-PEI were coated on silica nanoparticles through layer-by-layer (LbL) self-assembly with polyanions as monitored by dynamic light scattering, zeta-potential, and scanning electron microscopy. The thickness of the multilayer film was estimated from quartz crystal microbalance based on counting frequency change of each adsorbed layer. The magnetic relaxation of SiO2/(Gd-DTPA-PEl/polyanion), core-shell nanocomposite was tested at 1.5 T magnetic field in a clinical MRI scanner, and a 3-fold increase in T1 relaxivity to 15.1 Gd mM(-1)s(-1) was noticed comparing to Gd-DTPA small molecules. Dextran sulfate was coated as the outermost layer on the nanocomposite for better biocompatibility as verified by in vitro cytotoxicity studies. This formulation provides good signal intensity enhancement of mouse liver in vivo with only 1/25 dose of clinical standard at 30 and 60 minutes after intravenous injection. This sensitive imaging probe with unique core-shell structures may find broad applications in cellular and molecular imaging.

Self-assembly of magnetite nanocrystals with amphiphilic polyethylenimine: structures and applications in magnetic resonance imaging.

Controlled self-assembly of magnetic nanocrystals has important applications in biomedical fields. In this study, hydrophobic superparamagnetic iron oxide (SPIO) nanocrystals are self-assembled into small clusters (mean diameter <100 nm) in water phase with the help of partially alkylated hyper-branched polycation, polyethylenimine (PEI). The amphiphilic PEI can wrap one or multiple SPIO nanocrystals inside micelle cores depending on different polycation/SPIO mass ratios. These SPIO nanocrystal containing micelles are superparamagnetic at room temperature. At the magnetic field of 1.5 T, single SPIO nanocrystal containing micelles have a T2 relaxivity of 118 Fe mM(-1)s(-1), while multiple SPIO nanocrystals micelles have relatively higher T2 relaxivities up to 323 Fe mM(-1)s(-1). Next, these potential magnetic resonance imaging (MRI) probes are tested for labeling rabbit mesenchymal stem cells. The formulation with a low polymer/SPIO ratio (0.2) is biocompatible and effective in labeling. Under a clinical MRI scanner, 3 x 10(6) labeled cells in gelatin phantom present much darker images than controlled ones. The T2 relaxation rate of the labeled cells is approximately 74.2 s(-1), significantly higher than the control cells of approximately 3.4 s(-1). This new amphiphilic polycation/SPIO nanocomposite may provide opportunities in cell labeling and tracking their behaviors in vivo.

Linear and Core-Crosslinked Glycopolymer-Gadolinium Conjugates: Preparation and Their Behaviors as Nanoscale Magnetic Resonance Imaging Contrast Agents.

In this study, a linear glycopolymer-gadolinium conjugate (pGAEMA-DOTA-Gd) was prepared via the reversible addition fragmentation chain transfer (RAFT) polymerization. In addition, a crosslinked polymer-gadolinium conjugate (Crosslinked pGAEMA-DOTA-Gd) was prepared via a two-step RAFT polymerization approach, and its core was composed of the biodegradable oligopeptide GFLG. The features of the two glycopolymer-gadolinium conjugates as highly efficient and safe nanoscale contrast agents were discussed. These two glycopolymer-DOTA-Gd conjugates have aqueous dynamic particle sizes of 4.8 nm and 21.3 nm with neutral charges. Longitudinal relaxivity (r1) of these two glycopolymer-gadolinium conjugates were three to four times higher than that of clinical agent DTPA-Gd. Animal studies showed that pGAEMA-DOTA-Gd and Crosslinked pGAEMA-DOTA-Gd demonstrated higher signal intensity and the relative change in T1 value (ΔT1) in mouse liver and kidney, and prolonged blood circulation time compared to DTPA-Gd. Notably, the performance of the core-crosslinked glycopolymer conjugate was better than that of the linear polymer. Inductively coupled plasma mass spectrometry (ICP-MS) results showed that polymeric contrast agents, especially the crosslinked one, showed higher aggregation in mouse liver and kidney, and were mainly excreted through renal routes eventually. In vitro and in vivo toxicity studies in healthy mice including cytotoxicity, blood compatibility and systemic toxicity indicated the absence of significant toxicity. Therefore, the prepared glycopolymer-DOTA-Gd conjugates may have significant potential as highly efficient and safe nanoscale MRI contrast agents.

A selective fluorescent and colorimetric dual-responses chemosensor for streptomycin based on polythiophene derivative.

A colorimetric and fluorescent dual-responses chemosensor (PT3, a water-soluble polythiophene) for streptomycin was designed and synthesized. The structure of PT3 was characterized by using infrared spectroscopy, (1)H NMR and gel-permeation chromatography analyses. The conformational change of PT3 induced by streptomycin resulted in the red shift of absorption spectra and fluorescent quenching. Moreover, PT3 showed excellent selectivity for streptomycin over other antibiotics and biomolecules. PT3 could quantificationally detect streptomycin in the range of 2-70 µM with a detection limit of 0.2 µM (116 ppb), which is lower than the maximum residue limit defined by World Health Organization (200 ppb).

Preclinical evaluation of a urokinase plasminogen activator receptor-targeted nanoprobe in rhesus monkeys.

PURPOSE: To translate a recombinant peptide containing the amino-terminal fragment (ATF) of urokinase plasminogen activator receptor-targeted magnetic iron oxide (IO) nanoparticles (uPAR-targeted human ATF-IONPs) into clinical applications, we conducted a pilot study to evaluate the toxicity and pharmacokinetics of this nanoparticle in normal rhesus monkeys. METHODS: We assessed the changes in the following: magnetic resonance imaging (MRI) signals from pretreatment stage to 14 days posttreatment, serum iron concentrations from 5 minutes posttreatment to 12 weeks posttreatment, routine blood examination and serum chemistry analysis results from pretreatment stage to 12 weeks after administration, and results of staining of the liver with Perls' Prussian Blue and hematoxylin-eosin at 24 hours and 3 months posttreatment in two rhesus monkeys following an intravenous administration of the targeted nanoparticles either with a polyethylene glycol (ATF-PEG-IONP) or without a PEG (ATF-IONP) coating. RESULTS: The levels of alkaline phosphatase, alanine transaminase, and direct bilirubin in the two monkeys increased immediately after the administration of the IONPs but returned to normal within 20 days and stayed within the normal reference range 3 months after the injection. The creatinine levels of the two monkeys stayed within the normal range during the study. In addition, red blood cells, white blood cells, hemoglobin level, and platelets remained normal during the 3 months of the study. CONCLUSION: All of the results suggest that a transient injury in terms of normal organ functions, but no microscopic necrotic lesions, was observed at a systemic delivery dose of 5 mg/kg of iron equivalent concentration in the acute phase, and that no chronic toxicity was found 3 months after the injection. Therefore, we conclude that uPAR-targeted IONPs have the potential to be used as receptor-targeted MRI contrasts as well as theranostic agents for the detection and treatment of human cancers in future studies.