Harnessing the Potential of a Nitroreductase-Responsive Fluorescent Probe for the Diagnosis of Bacterial Keratitis.

Bacterial keratitis, an ocular emergency, is the predominant cause of infectious keratitis. However, diagnostic procedures for it are invasive, time-consuming, and expeditious, thereby limiting effective treatment for the disease in the clinic. It is imperative to develop a timely and convenient method for the noninvasive diagnosis of bacterial keratitis. Fluorescence imaging is a convenient and noninvasive diagnostic method with high sensitivity. In this study, a type of nitroreductase-responsive probe (NTRP), which responds to nitroreductase to generate fluorescence signals, was developed as an activatable fluorescent probe for the imaging diagnosis of bacterial keratitis. Imaging experiments both in vitro and in vivo demonstrated that the probe exhibited "turn-on" fluorescence signals in response to nitroreductase-secreting bacteria within 10 min. Furthermore, the fluorescence intensity reached its highest at 4 or 6 h in vitro and at 30 min in vivo when the excitation wavelength was set at 520 nm. Therefore, the NTRP has the potential to serve as a feasible agent for the rapid and noninvasive in situ fluorescence diagnosis of bacterial keratitis.

A Flexible Caterpillar-Like Gold Nanoparticle Assemblies with Ultrasmall Nanogaps for Enhanced Dual-Modal Imaging and Photothermal Therapy.

Gold nanoparticle (AuNP) assemblies (GNAs) have attracted attention since enhanced coupling plasmonic resonance (CPR) emerged in the nanogap between coupling AuNPs. For one dimensional GNAs (1D-GNAs), most CPR from the nanogaps could be easily activated by electromagnetic waves and generate drastically enhanced CPR because the nanogaps between coupling AuNPs are linearly distributed in the 1D-GNAs. The reported studies focus on the synthesis of 1D-GNAs and fundamental exploration of CPR. There are still problems which impede further applications in nanomedicine, such as big size (>500 nm), poor water solubility, and/or poor stability. In this study, a kind of 1D flexible caterpillar-like GNAs (CL-GNAs) with ultrasmall nanogaps, good water solubility, and good stability is developed. The CL-GNAs have a flexible structure that can randomly move to change their morphology, which is rarely reported. Numerous ultrasmall nanogaps (<1 nm) are linearly distributed along the structure of CL-GNAs and generate enhanced CPR. The toxicity assessments in vitro and vivo respectively demonstrate that CL-GNAs have a low cytotoxicity and good biocompatibility. The CL-GNAs can be used as an efficient photothermal agent for photothermal therapy, a probe for Raman imaging and photothermal imaging.

Correction: A Y1 receptor ligand synergized with a P-glycoprotein inhibitor improves the therapeutic efficacy of multidrug resistant breast cancer.

Correction for 'A Y1 receptor ligand synergized with a P-glycoprotein inhibitor improves the therapeutic efficacy of multidrug resistant breast cancer' by Yinjie Wang et al., Biomater. Sci., 2019, 7, 4748-4757, https://doi.org/10.1039/C9BM00337A.

A Y1 receptor ligand synergized with a P-glycoprotein inhibitor improves the therapeutic efficacy of multidrug resistant breast cancer.

Multidrug resistance (MDR) is one of the main reasons for the inefficiency of cancer chemotherapy. As a consequence of MDR, the expression level of membrane proteins might be changed, which can thus be used to develop a novel strategy for its treatment. Based on the high overexpression of Y1 receptor (Y1R) protein and P-glycoprotein (P-gp) in the multidrug resistant breast cancer cell line, a selective Y1R ligand [Asn6, Pro34]-NPY (AP) was employed to stabilize the chemotherapeutic drug doxorubicin (DOX) and P-gp inhibitor tariquidar (Tar) co-loaded nanomicelles at the physiological level. This also improved the targeted delivery of DOX and Tar into MCF-7/ADR cells. Co-delivered Tar further impedes the efflux of DOX and enhances its accumulation in the nuclei of drug resistant cancer cells, thereby inducing significant inhibition of cell growth. The synergistic effect of AP and Tar generates an excellent in vivo tumor targeting and antitumor efficacy of DOX with prolonged survival and minimized side effects, especially for liver metastasis. In general, Y1R as a novel target site and its selective ligand AP synergized with the P-gp inhibitor can be used for a more precise MDR breast cancer treatment.

Dual ATP and pH responsive ZIF-90 nanosystem with favorable biocompatibility and facile post-modification improves therapeutic outcomes of triple negative breast cancer in vivo.

Zeolitic imidazole frameworks (ZIFs) are becoming a notable nanosystem in biomedicine field, due to their unique properties of favorable biocompatibility, pH-responsive degradable structure and high drug loading. Compared with the increasing attention on ZIF-8 in cancer diagnosis and treatment, there is limited research about the bio-application of ZIF-90, especially its in vivo therapeutic efficacy and related toxicity. Here, we synthesize nano ZIF-90 through a fast self-assembling process, and the synthesized nano ZIF-90 is about 75 nm with a negative zeta potential, providing better mitochondria targetability, cell biocompatibility and in vivo survival rate comparing to nano ZIF-8. To further explore the applicability of ZIF-90 in cancer treatment, a facile post-modification is used to conjugate Y1 receptor ligand [Asn6, Pro34]-NPY (AP) on the surface of doxorubicin (DOX)-encapsulated nano ZIF-90. AP-ZIF-90 significantly reduces premature DOX release at physiological pH level, and triggers more effective and faster DOX release inside the tumor cells with dual responsive to high adenosine triphosphate (ATP) and low pH condition. Combining targeted delivery and dual responsive release of DOX significantly improves the therapeutic efficacy of AP-ZIF-90@DOX in MDA-MB-231 tumor bearing mouse, and results in 80% survival rate over 40 days of treatment. At the given dosage, nano ZIF-90 is with excellent biocompatibility in vivo, inducing minimal side effect on the liver and renal functions. Therefore, nano ZIF-90 combines with Y1 receptor ligand with favorable biocompatibility and dual responsiveness can be used as a promising nanosystem for targeted triple negative breast cancer treatment in vivo.

A Ultrasensitive Near-Infrared Fluorescent Probe Reveals Pyroglutamate Aminopeptidase 1 Can Be a New Inflammatory Cytokine.

Previous study showed that pyroglutamate aminopeptidase 1 (PGP-1) has a relationship with the immune response in cells. However, whether PGP-1 is involved in inflammatory response in vivo and can serve as a new inflammatory cytokine are still unclear. To address these issues, a new near-infrared fluorescent probe, which exhibits high selectivity and super sensitivity, is developed. With this probe, the up-regulation of PGP-1 (evidenced by western blot) in BALB/c mice legs and livers under the stimulation of two main immunopotentiators is revealed for the first time. The occurrence of inflammatory process (including tissue necrosis) in mice is determined by up-regulation of tumor necrosis factor-α and hematoxylin-eosin staining. Interestingly, it is revealed for the first time that knocking down PGP-1 leads to the weakness of inflammatory process in RAW264.7 cells. These new findings suggest that PGP-1 is indeed involved in inflammatory response in vivo and can be a new inflammatory cytokine.

A novel fibroblast activation protein-targeted near-infrared fluorescent off-on probe for cancer cell detection, in vitro and in vivo imaging.

A new hemicyanine-based fibroblast activation protein-targeted near-infrared fluorescent probe is designed and it shows high selectivity and sensitivity to cancer cell detection, and in vitro and in vivo imaging. This probe is successfully applied in fluorescence detection of living cells (with a detection limit of 1500 cells per mL). It is believed that many new functions or distributions of FAP could be discovered by this new probe later.

Fabrication of anti-fouling, anti-bacterial and non-clotting PVDF membranes through one step "outside-in" interface segregation strategy.

Herein we provide an "outside-in" interface segregation strategy to modify polyvinylidene fluoride (PVDF) membranes with anti-fouling, anti-bacterial and non-clotting performances for the first time. The coagulation bath composed of DMAc, water and synthesized copolymer was used to solidify and modify membrane during phase inversion process. The functional polymer e.g. poly(vinylpyrrolidone-vinyltriethoxysilane) (PVP-VTES), poly(dimethylamino ethyl methacrylate-vinyltriethoxysilane) (PDMAEMA-VTES), poly(acrylic acid-sodium p-styrene sulfonate-vinyltriethoxysilane) (PAA-SSNa-VTES) was segregated into membrane interface from outside coagulation bath. Further hydro-thermal reaction was implemented to crosslink the functional copolymer on membrane surface. The surface chemistry (ATR-FTIR, XPS) and physical properties (SEM, contact angle, flux) confirmed surface segregation of corresponding copolymer. The membranes showed excellent anti-fouling (BSA adsorption ∼19.1 µg/cm2), anti-bacterial (E. coli inhibition) and non-clotting (extended APTT, PT, TT and reduced FIB) performances respectively, which is clinically significant to hemocompatible membranes. The non-clotting membrane showed unprecedented prolonged activated partial thromboplastin time (APTT ∼ 53 s when sample area is minimized to 1/32 cm2). The thermodynamic segregation of the copolymer driven by surface tension and membrane gelation kinetics was investigated to understand the "outside-in" interface segregation route during phase inversion. Overall, the membrane fabrication and modification can be finished simultaneously in ∼2 min, demonstrating its potential in a continuous large scale production.

Neuropeptide Y Y1 receptor-mediated biodegradable photoluminescent nanobubbles as ultrasound contrast agents for targeted breast cancer imaging.

Targeted molecular imaging has attracted great attention in cancer diagnosis and treatment. However, most clinically used ultrasound contrast agents (UCAs) are non-targeted microbubbles seldom used for cancer imaging. Here, we fabricated fluorescent nanobubbles (NBs) by encapsulation of liquid tetradecafluorohexane (C6F14) within biodegradable photoluminescent polymers (BPLPs) through an emulsion-evaporation process and conjugation of PNBL-NPY ligand for specific targeting of Y1 receptors overexpressed in breast tumors. The developed PNBL-NPY modified NBs were uniform in size with good dispersibility and photostability, presenting good ultrasound enhancement. Further, in vitro and in vivo results indicated that the fabricated NBs exhibit high affinity and specificity to Y1 receptor-overexpressing breast cancer cells and tumors with minimal toxicity and damage to organs. Our developed PNBL-NPY-modified NBs are novel targeted UCAs for safe, efficient and specific targeted breast cancer imaging, and may provide a new nanoplatform for early cancer diagnosis and treatment in the future.

Silica-coated super-paramagnetic iron oxide nanoparticles (SPIONPs): a new type contrast agent of T1 magnetic resonance imaging (MRI).

Magnetic resonance imaging (MRI), a sophisticated promising three-dimensional tomographic noninvasive diagnostic technique, has an intrinsic advantage in safety compared with radiotracer and optical imaging modalities; however, MRI contrast agents are less sensitive than complexes used in other imaging techniques. Usually the clinically used Gd-based complexes MRI-T1 contrast agents are toxic; therefore, the demand for nontoxic novel T1-weighted MRI candidates with ultrasensitive imaging and advanced functionality is very high. In this research, silica-coated ultra-small monodispersed super-paramagnetic iron oxide nanoparticles were synthesized via a thermal decomposition method, which demonstrated themselves as a high performance T1-weighted MRI contrast agent for heart, liver, kidney and bladder based on in vivo imaging analyses. Transmission electron microscopy (TEM) results illustrated that the diameter of the SPIONPs was in the range of 4 nm and the average size of Fe3O4@SiO2 was about 30-40 nm. X-ray diffraction (XRD) and Raman spectroscopy analyses revealed the phase purity of the prepared SPIONPs. These magnetite nanoparticles exhibited a weak magnetic moment at room temperature because of the spin-canting effect, which promoted a high positive signal enhancement ability. MTT assays and histological analysis demonstrated good biocompatibility of the SPIONPs in vitro and in vivo. In addition, the silica-coated ultra-small (4 nm sized) magnetite nanoparticles exhibited a good r1 relaxivity of 1.2 mM-1 s-1 and a low r2/r1 ratio of 6.5 mM-1 s-1. In vivo T1-weighted MR imaging of heart, liver, kidney and bladder in mice after intravenous injection of nanoparticles further verified the high sensitivity and biocompatibility of the as-synthesized magnetite nanoparticles. These results reveal silica-coated SPIONPs as a promising candidate for a T1 contrast agent with extraordinary capability to enhance MR images.

Neuropeptide Y Y1 receptors mediate [corrected] targeted delivery of anticancer drug with encapsulated nanoparticles to breast cancer cells with high selectivity and its potential for breast cancer therapy.

By enabling nanoparticle-based drug delivery system to actively target cancer cells with high selectivity, active targeted molecules have attracted great attention in the application of nanoparticles for anticancer drug delivery. However, the clinical application of most active targeted molecules in breast cancer therapy is limited, due to the low expression of their receptors in breast tumors or coexpression in the normal and tumor breast tissues. Here, a neuropeptide Y Y1 receptors ligand PNBL-NPY, as a novel targeted molecule, is conjugated with anticancer drug doxorubicin encapsulating albumin nanoparticles to investigate the effect of Y1 receptors on the delivery of drug-loaded nanoparticles to breast cancer cells and its potential for breast cancer therapy. The PNBL-NPY can actively recognize and bind to the Y1 receptors that are significantly overexpressed on the surface of the breast cancer cells, and the drug-loaded nanoparticles are delivered directly into the cancer cells through internalization. This system is highly selective and able to distinguish the breast cancer cells from the normal cells, due to normal breast cells that express Y2 receptors only. It is anticipated that this study may provide a guidance in the development of Y1 receptor-based nanoparticulate drug delivery system for a safer and more efficient breast cancer therapy.

Improved double emulsion technology for fabricating autofluorescent microcapsules as novel ultrasonic/fluorescent dual-modality contrast agents.

The aim of this study is to explore an improved double emulsion technology with in situ reaction of lysine (Lys) and glutaraldehyde (GA) for fabricating autofluorescent Lys-poly(lactic-co-glycolic acid)-GA (Lys-PLGA-GA) microcapsules as novel ultrasonic/fluorescent dual-modality contrast agents. Scanning electron microscope (SEM) and static light scattering (SLS) results show that 80% of the Lys-PLGA-GA microcapsules are larger than 1.0 µm and 90% of them are smaller than 8.9 µm. SEM and laser confocal scanning microscope (LCSM) data demonstrate that the structure of our Lys-PLGA-GA microcapsules is hollow. Compared with the FT-IR spectrum of PLGA microcapsules, a new peak at 1,644 cm(-1) in that of Lys-PLGA-GA microcapsules confirms the formed Schiff base in Lys-PLGA-GA microcapsules. LCSM images and fluorescence spectra show that our Lys-PLGA-GA microcapsules exhibit bright and stable autofluorescence without conjugation to any fluorescent agent, which can be ascribed to the n-π transitions of the CN bonds in the formed Schiff base. Our autofluorescent Lys-PLGA-GA microcapsules might have more wide applications than traditional fluorescent dyes because their excitation and emission spectra are both broad. The fluorescence intensity can also be tuned by the feeding amount of Lys and GA. The MTT assays reveal that the autofluorescent microcapsules are biocompatible. The results of fluorescent imaging in cells and in vitro ultrasonic imaging demonstrate the feasibility of our autofluorescent Lys-PLGA-GA microcapsules as ultrasonic/fluorescent dual-modality contrast agents. This novel ultrasonic/fluorescent dual-modality contrast agent might have potential for a variety of biological and medical applications.

Multifunctional Fe3O4-TiO2 nanocomposites for magnetic resonance imaging and potential photodynamic therapy.

Multifunctional Fe(3)O(4)-TiO(2) nanocomposites with Janus structure for magnetic resonance imaging (MRI) and potential photodynamic therapy (PDT) were synthesized, in which Fe(3)O(4) was used as a MRI contrast agent and TiO(2) as an inorganic photosensitizer for PDT. Their morphology, structure, and MRI and PDT performance were characterized, respectively. Moreover, the location of Fe(3)O(4)-TiO(2) nanocomposites in MCF-7 cells was also investigated by the staining of Prussian blue and alizarin red, respectively. The results showed that the as-prepared Fe(3)O(4)-TiO(2) nanocomposites had good T(2)-weighted MRI performance, and the MCF-7 cells incubated with nanocomposites could be killed under the irradiation of UV light. Compared with traditional organic photosensitizers, TiO(2) inorganic photosensitizers could have more stable PDT performance due to their nanoscale size and anti-photodegradable stability. Therefore, the as-prepared Fe(3)O(4)-TiO(2) nanocomposites could have potential applications as a new kind of multifunctional agent for both MRI and PDT.

Biocompatible composite nanoparticles with large longitudinal relaxivity for targeted imaging and early diagnosis of cancer.

Early diagnosis of cancer greatly increases the chances of successful treatment by radical resection. The sensitivity of magnetic resonance imaging (MRI) techniques for detecting early stage tumors can be increased with the assistance of a positive MRI contrast agent. However, the traditional positive MRI contrast agents, such as Gd-chelates and Gd-based inorganic nanoparticles, are often limited by their cytotoxicity and low specificity. Here, we propose a new design of MRI contrast agent based on gadolinium oxide nanocrystals (GON) for targeted imaging and cancer early diagnosis with good biocompatibility. The GON were prepared using a polyol method and then encapsulated into albumin nanoparticles (AN), which were cross-linked with glutaraldehyde and found to exhibit bright and stable autofluorescence without conjugation to any fluorescent agent. After that, a target molecule, folic acid (FA), was conjugated onto the surface of the GON-loaded AN (GON-AN) to construct a GON-AN-FA composite. The as-prepared nanoparticles are biocompatible and stable in serum. The results of MRI relaxation studies show that the longitudinal relaxivities (r1) of GON-AN (11.6 mM-1 s-1) and GON-AN-FA (10.8 mM-1 s-1) are much larger than those of traditional positive MRI contrast agents, such as Magnevist (3.8 mM-1 s-1). The results of cell viability assays indicate that GON-AN and GON-AN-FA are almost non-cytotoxic. Furthermore, the specificities of GON-AN and GON-AN-FA were evaluated with two kinds of cancer cells which overexpress folate receptor alpha (FRα). The results reinforce that the autofluorescent GON-AN-FA is able to target cancer cells via recognition of the ligand FA and the receptor FRα. Therefore, our autofluorescent GON-AN-FA possessing a large longitudinal relaxivity and good biocompatibility represents a significant advance for the targeted imaging and early diagnosis of cancer.