Ultrasmall Magnetically Engineered Ag2Se Quantum Dots for Instant Efficient Labeling and Whole-Body High-Resolution Multimodal Real-Time Tracking of Cell-Derived Microvesicles.

Cell-derived microvesicles (MVs) are natural carriers that can transport biological molecules between cells, which are expected to be promising delivery vehicles for therapeutic purposes. Strategies to label MVs are very important for investigation and application of MVs. Herein, ultrasmall Mn-magnetofunctionalized Ag2Se quantum dots (Ag2Se@Mn QDs) integrated with excellent near-infrared (NIR) fluorescence and magnetic resonance (MR) imaging capabilities have been developed for instant efficient labeling of MVs for their in vivo high-resolution dual-mode tracking. The Ag2Se@Mn QDs were fabricated by controlling the reaction of Mn(2+) with the Ag2Se nanocrystals having been pretreated in 80 °C NaOH solution, with an ultrasmall size of ca. 1.8 nm, water dispersibility, high NIR fluorescence quantum yield of 13.2%, and high longitudinal relaxivity of 12.87 mM(-1) s(-1) (almost four times that of the commercial contrast agent Gd-DTPA). The ultrasmall size of the Ag2Se@Mn QDs enables them to be directly and efficiently loaded into MVs by electroporation, instantly and reliably conferring both NIR fluorescence and MR traceability on MVs. Our method for labeling MVs of different origins is universal and free of unfavorable influence on intrinsic behaviors of MVs. The complementary imaging capabilities of the Ag2Se@Mn QDs have made the long-term noninvasive whole-body high-resolution dual-mode tracking of MVs in vivo realized, by which the dynamic biodistribution of MVs has been revealed in a real-time and in situ quantitative manner. This work not only opens a new window for labeling with QDs, but also facilitates greatly the investigation and application of MVs.

Fluorescence Detection of H5N1 Virus Gene Sequences Based on Optical Tweezers with Two-Photon Excitation Using a Single Near Infrared Nanosecond Pulse Laser.

We present an analytical platform by combining near-infrared optical tweezers with two-photon excitation for fluorescence detection of H5N1 virus gene sequences. A heterogeneous enrichment strategy, which involved polystyrene (PS) microsphere and quantum dots (QDs), was adopted. The final hybrid-conjugate microspheres were prepared by a facile one-step hybridization procedure by using PS microspheres capturing target DNA and QDs tagging, respectively. Quantitative detection was achieved by the optical tweezers setup with a low-cost 1064 nm nanosecond pulse laser for both optical trapping and two-photon excitation for the same hybrid-conjugate microsphere. The detection limits for both neuraminidase (NA) gene sequences and hemagglutinin (HA) gene sequences are 16-19 pM with good selectivity for one-base mismatch, which is approximately 1 order of magnitude lower than the most existing fluorescence-based analysis method. Besides, because of the fact that only signal from the trapped particle is detected upon two-photon excitation, this approach showed extremely low background in fluorescence detection and was successfully applied to directly detect target DNA in human whole serum without any separation steps and the corresponding results are very close to that in buffer solution, indicating the strong anti-interference ability of this method. Therefore, it can be expected to be an emerging alternative for straightforward detecting target species in complex samples with a simple procedure and high-throughput.

Enhanced delivery of theranostic liposomes through NO-mediated tumor microenvironment remodeling.

Highly efficient delivery of nanoagents to the tumor region remains the primary challenge for cancer nanomedicine. Herein, we propose a NO-mediated tumor microenvironment (TME) remodeling strategy for the high-efficient delivery of nanoagents into tumor. Quantum dots (QDs) with bright fluorescence in the near-infrared IIb (NIR-IIb, 1500-1700 nm) window and high photothermal conversion efficiency were encapsulated into liposomes for the imaging-guided photothermal therapy (PTT) of tumor. The fabrication of PEG and arginine-glycine-aspartate (RGD) peptide on liposomes ensured the prolonged circulation in vivo and active targeting to tumor. Moreover, the loading of a natural NO generator L-arginine in liposomes realized the continuous generation of NO in the acidic TME. By co-localization fluorescence imaging and western blot of tumor tissue, we confirmed that the release of NO activated the expression of metalloproteinases in TME and further degraded Collagen I in the peripheral region of the tumor, thus removing the barrier for the permeation of liposomes. Attributed to the enhanced accumulation of liposomes inside the tumor, NIR IIb imaging-guided PTT was achieved with remarkable therapeutic efficacy.

An Ultra-Stable, Oxygen-Supply Nanoprobe Emitting in Near-Infrared-II Window to Guide and Enhance Radiotherapy by Promoting Anti-Tumor Immunity.

Currently, radiotherapy (RT) is the main method for cancer treatment. However, the hypoxic environment of solid tumors is likely to cause resistance or failure of RT. Moreover, high-dose radiation may cause side effects to surrounding normal tissues. In this study, a new type of nanozyme is developed by doping Mn (II) ions into Ag2 Se quantum dots (QDs) emitting in the second near-infrared window (NIR-II, 1000-1700 nm). Through the catalysis of Mn (II) ions, the nanozymes can trigger the rapid decomposition of H2 O2 and produce O2 . Conjugated with tumor-targeting arginine-glycine-aspartate (RGD) tripeptides and polyethylene glycol (PEG) molecules, the nanozymes are then constructed into in vivo nanoprobes for NIR-II imaging-guided RT of tumors. Owing to the radiosensitive activity of the element Ag, the nanoprobes can promote radiation energy deposition. The specific tumor-targeting and NIR-II emitting abilities of the nanoprobes facilitate the precise tumor localization, which enables precise RT with low side effects. Moreover, their ultra-stability in the living body ensures that the nanoprobes continuously produce oxygen and relieve the hypoxia of tumors to enhance RT efficacy. Guided by real-time and high-clarity imaging, the nanoprobe-mediated RT promotes anti-tumor immunity, which significantly inhibits the growth of tumors or even cures them completely.

Cytocompatibility and Bone-Formation Potential of Se-Coated 316L Stainless Steel with Nano-Pit Arrays.

316L stainless steel is still widely applied in joint replacement and orthopedic surgery due to its mechanical properties, corrosion resistance and relatively low price. In this study, electrochemical oxidation and nanoscale coating were used to fabricate Se-coated 316L stainless steel with nano-pit arrays to enhance its surface characteristics, biocompatibility and osseointegration ability. The modified 316L stainless steel was tested via field emission scanning microscopy (FESEM), energy-dispersive X-ray spectrometry (EDS), X-ray photoelectron spectroscopy (XPS) and Se release studies. The results of this study showed that the nano-pit arrays were 50 nm in diameter and that the Se coating consisted primarily of elemental Se and exhibited sustained release. The biological response of the samples was evaluated using in vitro rat bone marrow mesenchymal stem cell (rBMSCs) experiments and in vivo animal experiments. The modified 316L stainless steel displays enhanced abilities of cell adhesion, proliferation and osteogenic activity, as shown by FESEM, CCK-8 assay, immunofluorescence microscopy (IF) and alkaline phosphatase (ALP) activity assay in vitro and additional new bone formation in vivo, indicating its outstanding cytocompatibility and osteogenic differentiation ability. More importantly, the Se coating can upregulate gene expression of OPN, RUNX-2 and ALP, indicating that the nano-Se-coated 316L stainless steel with nano-pit arrays is a promising biomedical material for implants in orthopedic or dental clinic applications.

Odontogenic ameloblast-associated protein (ODAM) inhibits human colorectal cancer growth by promoting PTEN elevation and inactivating PI3K/AKT signaling.

Odontogenic ameloblast-associated protein (ODAM), an acidic matricellular protein, has been implicated in several epithelial neoplasms. However, its biological functions and molecular mechanisms in cancer progression, particular colorectal carcinoma (CRC), remain unknown. Here we demonstrated that ODAM was significantly down-regulated in CRC tissues compared with their normal counterparts. Then, we established that ODAM expression level was closely correlated with CRC development and patient prognosis. The abnormal expression of ODAM dramatically affected CRC cell growth in vitro and in vivo. We further revealed that the inhibitory effects of ODAM on CRC cell growth were associated with PTEN elevation and PI3K/AKT signaling inactivation. Furthermore, we determined that silencing of PTEN expression yielded recovery of AKT activity in ODAM-expressing CRC cells. Our study suggests matricellular protein ODAM may serve as a novel prognostic marker and act as a CRC growth suppressor.

Intermediate-dominated controllable biomimetic synthesis of gold nanoparticles in a quasi-biological system.

A new biomimetic strategy of creating a quasi-biological system (an aqueous solution containing electrolytes, peptide, enzyme and coenzyme) for the preparation of gold nanoparticles with uniform and tunable sizes has been put forward and validated, adopting environmentally-friendly reducing agents and a biocompatible capping ligand in aqueous solution at room temperature. The biomimetic synthetic route has the characteristics for good stability of the resulting AuNPs capped with glutathione via strong Au-S bond in aqueous solution, an appropriate composition of the intermediate with a redox potential favorable for the biomimetic reduction under mild conditions, suitable pH values to adjust the rate of the reduction, and the addition of enzyme catalyzing the reduction. By only adjusting the concentration of the reducing agent NADPH, a series of AuNPs with narrow size-distribution could be controllably synthesized. This method of rational utilization of biological processes could provide a new way for the sustainable development of nanotechnology.

Visualized investigation of yeast transformation induced with Li+ and polyethylene glycol.

The effects of Li(+) and polyethylene glycol (PEG) on the genetic transformation of Saccharomyces cerevisiae were investigated by using fluorescence microscopy (FM) to visualize the binding of plasmid DNA labeled with YOYO-1 to the surface of yeast cells, scanning electron microscopy (SEM) and atomic force microscopy (AFM) to image the change in surface topography of yeast cells, coupled with transformation frequency experiments. The results showed that under the same conditions, the transformation frequencies of yeast protoplasts were much higher than those of intact yeast cells. PEG was absolutely required for the binding of DNA to the surface of intact yeast cells or yeast protoplasts, and had no effect on the surface topography of intact yeast cells or yeast protoplasts. In the presence of PEG, Li(+) could greatly enhance the binding of plasmid DNA to the surface of intact yeast cells, increase their transformation frequency, and affect their surface topography. On the other hand, no effect on the DNA binding to the surface of protoplasts and no increase in the number of transformants and no surface topography changes were found upon the treatment with Li(+) to protoplasts. In the present work, the effects of Li(+) and PEG on yeast genetic transformation were directly visualized, rather than those deduced from the results of transformation frequencies. These results indicate that cell wall might be a barrier for the uptake of plasmid DNA. Li(+) could increase the permeability of yeast cell wall, then increase the exposed sites of DNA binding on intact yeast cells. The main role of PEG was to induce DNA binding to cell surface.