Remodeling of tumor microenvironment for enhanced tumor chemodynamic/photothermal/chemo-therapy.

The anticancer treatment is largely affected by the microenvironment of the tumors, which not only resists the tumors to the thermo/chemo-therapy, but also promotes their growth and invasion. In this work, the angiogenesis factor is balanced by combining with the breathing hyperoxygen, for regulating the tumor microenvironment and also for relieving hypoxia and high tissue interstitial pressure, which promote drug delivery to tumor tissues by increasing the in vivo perfusion and reversing the immunosuppressive tumor. In addition, the designed multifunctional nanoparticles have a great potential for applications to the tumor dual-mode imaging including magnetic resonance (MR) and photoacoustic (PA) imaging. This work proposes a promising strategy to enhance the thermo/chemo-therapy efficacy by remodeling the tumor microenvironment, which would provide an alternative to prolong the lifetime of tumor patients.

Transesterification Synthesis of Chloramphenicol Esters with the Lipase from Bacillus amyloliquefaciens.

This work presents a synthetic route to produce chloramphenicol esters by taking advantage the high enantio- and regio-selectivity of lipases. A series of chloramphenicol esters were synthesized using chloramphenicol, acyl donors of different carbon chain length and lipase LipBA (lipase cloned from Bacillus amyloliquefaciens). Among acyl donors with different carbon chain lengths, vinyl propionate was found to be the best. The influences of different organic solvents, reaction temperature, reaction time, enzyme loading and water content on the synthesis of the chloramphenicol esters were studied. The synthesis of chloramphenicol propionate (0.25 M) with 4.0 g L-1 of LipBA loading gave a conversion of ~98% and a purity of ~99% within 8 h at 50 °C in 1,4-dioxane as solvent. The optimum mole ratio of vinyl propionate to chloramphenicol was increased to 5:1. This is the first report of B. amyloliquefaciens lipase being used in chloramphenicol ester synthesis and a detailed study of the synthesis of chloramphenicol propionate using this reaction. The high enzyme activity and selectivity make lipase LipBA an attractive catalyst for green chemical synthesis of molecules with complex structures.

Tumor-Penetrating Peptide-Modified DNA Tetrahedron for Targeting Drug Delivery.

DNA self-assembling nanostructure has been considered as a promising candidate as a drug delivery vehicle because of its compactness, mechanical stability, and noncytotoxicity. In this work, we developed functional, multiform DNA nanostructures by appending a tumor-penetrating peptide to tetrahedral DNA nanostructure (p-TDN). This functional structure is able to efficiently increase the rate of uptake of glioblastoma cell U87MG compared with the DNA tetrahedron and the double-stranded DNA structures. We found that the DNA tetrahedron plays the main role in the endocytosis of U87MG cells, whereas the tumor-penetrating peptide could also bind to transmembrane glycoprotein neuropilin-1 and mediate the endocytosis of the p-TDN nanostructure. Moreover, given the high efficiency of the growth inhibitory effect of the p-TDN loading doxorubicin hydrochloride, the p-TDN distinguishes itself as a promising candidate as an effective delivery carrier.

Aptamer-Functionalized and Backbone Redox-Responsive Hyperbranched Polymer for Targeted Drug Delivery in Cancer Therapy.

A novel type of backbone redox-responsive hyperbranched poly(2-((2-(acryloyloxy)ethyl)disulfanyl)ethyl 4-cyano-4-(((propylthio)carbonothioyl)-thio)-pentanoate-co-poly(ethylene glycol) methacrylate) (HPAEG) has been designed and prepared successfully via the combination of reversible addition-fragmentation chain-transfer (RAFT) polymerization and self-condensing vinyl polymerization (SCVP). Owing to the existence of surface vinyl groups, HPAEG could be efficiently functionalized by DNA aptamer AS1411 via Michael addition reaction to obtain an active tumor targeting drug delivery carrier (HPAEG-AS1411). The amphiphilic HPAEG-AS1411 could form nanoparticles by macromolecular self-assembly strategy. Cell Counting Kit-8 (CCK-8) assay illustrated that HPAEG-AS1411 nanoparticles had low cytotoxicity to normal cell line. Flow cytometry and confocal laser scanning microscopy (CLSM) results demonstrated that HPAEG-AS1411 nanoparticles could be internalized into tumor cells via aptamer-mediated endocytosis. Compared with pure HPAEG nanoparticles, HPAEG-AS1411 nanoparticles displayed enhanced tumor cell uptake. When the HPAEG-AS1411 nanoparticles loaded with anticancer drug doxorubicin (DOX) were internalized into tumor cells, the disulfide bonds in the backbone of HPAEG-AS1411 were cleaved by glutathione (GSH) in the cytoplasm, so that DOX was released rapidly. Therefore, DOX-loaded HPAEG-AS1411 nanoparticles exhibited a high tumor cellular proliferation inhibition rate and low cytotoxicity to normal cells. This aptamer-functionalized and backbone redox-responsive hyperbranched polymer provides a promising platform for targeted drug delivery in cancer therapy.

Self-assembly of DNA-based drug delivery nanocarriers with rolling circle amplification.

DNA nanostructures have recently emerged as a type of drug delivery nanocarriers due to their suitable sizes, well-defined structures and low-toxicity. Here, we present a protocol for the assembly of DNA nanoribbon structures with rolling circle amplification (RCA) and delivery of CpG oligonucleotide. DNA nanoribbons with different dimensions and patterns were assembled with long RCA strands and several short staples. Significantly, we demonstrated they exhibited high-efficiency cellular uptake and improved immunostimulatory activity compared with ss- or ds- DNA.

An olive-shaped SnO2 nanocrystal-based low concentration H2S gas sensor with high sensitivity and selectivity.

Olive-shaped SnO2 nanocrystals were synthesized successfully via a facile hydrothermal route, using tin dichloride hydrate, oxalic acid dihydrate and polyvinylpyrrolidone as reaction precursors, and showed great potential in the large-scale preparation of SnO2 nanocrystals. The prepared SnO2 nanocrystals were characterized using XRD, XPS, SEM, TEM and HRTEM, and showed well-defined olive-shaped tetragonal single-crystals with irregular exposed facets. The growth mechanism of the olive-shaped SnO2 nanocrystals was considered after investigating the experimental conditions and reaction time. Due to the abundant active sites on the irregular surfaces, the gas sensing performance of the prepared SnO2 nanocrystals exhibited great gas sensing properties, including high sensitivity, selectivity and stability towards H2S with a very low detection limit (less than 0.5 ppm), revealing their great potential in commercial applications for H2S gas detection.

Growth and origami folding of DNA on nanoparticles for high-efficiency molecular transport in cellular imaging and drug delivery.

A novel three-dimensional (3D) superstructure based on the growth and origami folding of DNA on gold nanoparticles (AuNPs) was developed. The 3D superstructure contains a nanoparticle core and dozens of two-dimensional DNA belts folded from long single-stranded DNAs grown in situ on the nanoparticle by rolling circle amplification (RCA). We designed two mechanisms to achieve the loading of molecules onto the 3D superstructures. In one mechanism, ligands bound to target molecules are merged into the growing DNA during the RCA process (merging mechanism). In the other mechanism, target molecules are intercalated into the double-stranded DNAs produced by origami folding (intercalating mechanism). We demonstrated that the as-fabricated 3D superstructures have a high molecule-loading capacity and that they enable the high-efficiency transport of signal reporters and drugs for cellular imaging and drug delivery, respectively.

Hybridization chain reaction amplification of microRNA detection with a tetrahedral DNA nanostructure-based electrochemical biosensor.

There remains a great challenge in the sensitive detection of microRNA because of the short length and low abundance of microRNAs in cells. Here, we have demonstrated an ultrasensitive detection platform for microRNA by combining the tetrahedral DNA nanostructure probes and hybridization chain reaction (HCR) amplification. The detection limits for DNA and microRNA are 100 aM and 10 aM (corresponding to 600 microRNAs in a 100 µL sample), respectively. Compared to the widely used supersandwich amplification, the detection limits are improved by 3 orders of magnitude. The uncontrolled surface immobilization and consumption of target molecules that limit the amplification efficiency of supersandwich are eliminated in our platform. Taking advantage of DNA nanotechnology, we employed three-dimensional tetrahedral DNA nanostructure as the scaffold to immobilize DNA recognition probes to increase the reactivity and accessibility, while DNA nanowire tentacles are used for efficient signal amplification by capturing multiple catalytic enzymes in a highly ordered way. The synergetic effect of DNA tetrahedron and nanowire tentacles have proven to greatly improve sensitivity for both DNA and microRNA detection.

Molecular logic gates on DNA origami nanostructures for microRNA diagnostics.

Molecular computing holds great promise for diagnosis and treatment of diseases at the molecular level; nevertheless, designing molecular logic gates to operate programmably and autonomously for molecular diagnostics still remains challenging. We designed logic gates on DNA Origami for microRNA analysis. As a demonstration, two indicators of heart failure, microRNA-21 and microRNA-195, were selected as the logic inputs. The logic gates contain two main modules: computation module and output module, performing in a single DNA Origami nanostructure. The computation module recognizes disease indicators, while the output module display different nanoscale symbols, "+" (positive) or "-" (negative), depending on the computing results. We demonstrated that the molecular logic gates worked well with single and two input combinations.

Long-term effects of nanoparticles on nutrition and metabolism.

Nanoparticles have shown great potential in biological and biomedical applications due to their distinct physical and chemical properties. In the meanwhile, the biosafety of nanoparticles has also raised intense concerns worldwide. To address such concerns, great efforts have been made to examine short-term effects of nanoparticles on cell survival and proliferation. More recently, exploration of long-term effects of nanomaterials, particularly those with promising biomedical applications in vivo, has aroused significant interest. For example, gold nanoparticles (AuNPs) are generally considered non-toxic to cell growth, whereas recent studies suggest that AuNPs might have long-term effects on cellular metabolism and energy homeostasis. In this Review, recent advances in this direction are summarized. Further, possible mechanisms under which nanoparticles regulate metabolic signaling pathways, potential long-term effects on cellular anabolic or catabolic processes, and their implications in human health and metabolic disorders are discussed.

Self-assembly of regenerated silk fibroin from random coil nanostructures to antiparallel ß-sheet nanostructures.

In this work, we studied the effects of incubation concentration and time on the self-assembly behaviors of regenerated silk fibroin (RSF). Our results showed the assembly ways of RSF were concentration-dependent and there were four self-assembly ways of RSF: (i) At relatively low concentration (≤0.015%), RSF molecules assembled into protofilaments (random coil), and then the thickness decreased and the secondary conformation changed to antiparallel ß-sheet; (ii) at the concentration of 0.015%, RSF molecules assembled into protofilaments (random coil), and then assembled into protofibrils (antiparallel ß-sheet). The protofibrils experienced the appearance and disappearance of phase periodic intervals in turn; (iii) at the concentration of 0.03%, RSF molecules assembled into bead-like oligomers (random coil), and then assembled into protofibrils (antiparallel ß-sheet), and finally the height and phase periodic intervals of RSF protofibrils disappeared in turn; and (iv) at the relatively high concentration (≥0.15%), RSF molecules assembled into protofilaments (random coil), then aggregated into blurry cuboid-like micelles (random coil), and finally self-arranged to form smooth and clear cuboid-like micelles (antiparallel ß-sheet). These results provide useful insights into the process by which the RSF molecules self-assemble into protofilaments, protofibrils and micelles. Furthermore, our work will be beneficial to basic understanding of the nanoscale structure formations in different silk-based biomaterials.

Tumor-penetrating peptide mediation: an effective strategy for improving the transport of liposomes in tumor tissue.

Currently, the inefficient transport of liposomes in tumor tissue hinders their clinical application. Tumor-penetrating peptides (TPP) are a series of targeting peptides with the function of penetrating tumor blood vessels and tumor stroma. This work aimed to improve the penetration of liposomes in tumor tissues by TPP modification, thereby enhancing the antitumor effect. First, RPARPAR, a TPP, was modified to the surface of liposomes loaded with doxorubicin. The RPARPAR-modified liposomes (RPA-LP) and unmodified liposomes (LP) showed spherical morphology with average sizes about 90 nm. RPA-LP exhibited remarkably increased cellular accumulation by PC-3 tumor cells than LP as evidenced by the cellular uptake test. The in vivo imaging study confirmed that RPARPAR modification significantly increased the liposome accumulation in subcutaneous tumor tissues. RPA-LP could penetrate through tumor blood vessels and tumor stroma and into the deep tumor tissues as evidence by the immunofluorescence staining analysis. The cytotoxicity of RPARPAR-modified doxorubicin liposomes (RPA-LP-DXR) is considerably increased compared with that of doxorubicin liposomes (LP-DXR). The RPA-LP-DXR also showed significantly (p < 0.005) stronger growth-inhibiting effect on tumor than LP-DXR, possibly due to the tumor-penetrating ability of RPA-LP and targeted killing of tumor cells. This study proved that TPP mediation may be an effective strategy for improving the transport of liposomes in tumor tissue.

Catalytic oxidation of low-concentration CO at ambient temperature over supported Pd-Cu catalysts.

The CO catalytic oxidation at ambient temperature and high space velocity was studied over the Pd-Cu/MOx (MOx = TiO2 and AI203) catalysts. The higher Brunauer-Emmett-Teller area surface of the A1203 support facilitates the dispersion of Pd2+ species, and the presence of Cu2Cl(OH)3 accelerates the re-oxidation of Pd0 to Pd2+ over the Pd-Cu/Al203 catalyst, which contributed to better performance of CO catalytic oxidation. The poorer activity of the Pd-Cu/TiO2 catalyst was attributed to the lower dispersion of Pd2+ species because of the less surface area and the non-formation of Cu2CI(OH)3 species. The presence of saturated moisture showed a negative effect on CO conversion over the two catalysts. This might be because of the competitive adsorption, the formation of carbonate species and the transformation of Cu2CI(OH)3 to inactive CuCI over the Pd-Cu/AI2O3 catalyst, which facilitates the aggregation of PdO species over the Pd-Cu/TiO2 catalyst under the moisture condition.

Enhanced visible-light activation of persulfate for the removal of RhB by supported nano-(C,N,B)-tridoped TiO2/copper foam mixed-crystals.

Photocatalytic persulfate activation by TiO2 and its application in sewage treatment have aroused great interest because of its high decontamination ability and strong adaptability, but the low light energy utilization rate and poor recycling of TiO2 limited its practical application. Herein, by using C-, N-, and B-modified TiO2 and immobilizing it on copper foam, we prepared a new and efficient (C,N,B)-TiO2/copper foam photocatalyst with enhanced visible-light activation performance of persulfate for the removal of RhB. It almost completely degraded RhB within 15 min of UV-vis light photocatalysis-assisted persulfate oxidation reaction with TOC removal of 53.17% in 30 min and presented the excellent long-term recyclability and stability, which is much better or comparative than those photocatalysts in the related literatures. (C,N,B)-TiO2/copper foam exhibited the largest apparent rate constant (0.149 min-1), 1.16 times higher than (C,N,B)-TiO2 (0.128 min-1), and 2.40 times higher than that of TiO2 (0.062 min-1), respectively. C,N,B doping modified the crystalline phase of TiO2, narrowed its band gap, and reduced charge-carrier recombination rate. These, together with the synergistic effect between photocatalysis and persulfate activation for enhancing generation of active species, jointly promoted the performance enhancement of TiO2. The 1O2 was the primary oxidation active species for the degradation of RhB, and the radical species (SO4•-, •O2-, and •OH) could further accelerate the photocatalytic activation of persulfate reaction.

Carbon nanotubes multifunctionalized by rolling circle amplification and their application for highly sensitive detection of cancer markers.

There are still challenges for the development of multifunctional carbon nanotubes (CNTs). Here, a multiwalled carbon nanotube (MWCNT)-based rolling circle amplification system (CRCAS) is reported which allows in situ rolling circle replication of DNA primer on the surface of MWCNTs to create a long single-strand DNA (ssDNA) where a large number of nanoparticles or proteins could be loaded, forming a nano-biohybridized 3D structure with a powerful signal amplification ability. In this strategy, the binding ability of proteins, hybridization, replication ability of DNA, and the catalytical ability of enzymes are integrated on a single carbon nanotube. The CRCAS is then used to develop colorimetric and chemiluminescent assays for the highly sensitive and specific detection of cancer protein markers, alpha-fetoprotein (AFP) and prostate specific antigen (PSA). The colorimetric CRCAS assay is 4000 times more sensitive than a conventional enzyme-linked immunosorbent assay (ELISA), and its concentration range is 10,000 times wider. Control experiments show that as low as 10 pg mL⁻¹ AFP or PSA could be detected even in the presence of interfering protein markers with a more than 105-fold greater concentration in the sample, demonstrating the high specificity of the CRCAS assay. The limit of detection of the chemiluminescent CRCAS assays for AFP and PSA are 5 fg mL⁻¹ (70 aM) and 10 fg mL⁻¹ (0.29 fM), respectively, indicating that the sensitivity is much higher than that of the colorimetric CRCAS assay. Importantly, CRCAS works well with real biological samples.

Tumor-penetrating peptide functionalization enhances the anti-glioblastoma effect of doxorubicin liposomes.

The targeted therapeutic effect of nano drug delivery system for glioblastoma has been hampered by the weak enhanced permeability and retention (EPR) effect of glioblastoma and the low delivering efficiency of NDDS in glioblastoma tissue. In this study, a tumor-penetrating peptide (RGERPPR), the specific ligand of neuropilin-1 overexpressed on glioblastoma and endothelial cells, was used as a targeting moiety to enhance the anti-glioblastoma effect of doxorubicin liposomes. Firstly, RGERPPR-PEG-DSPE was synthesized and used to prepare the RGERPPR peptide-functionalized liposomes (RGE-LS), which showed vesicle sizes of around 90 nm and narrow size distributions. The cellular uptake and in vivo near-infrared fluorescence imaging test displayed that RGE-LS exhibited increased uptake by glioblastoma cells and intracranial glioblastoma tissues. The cytotoxicity assay and anti-glioblastoma study proved that RGERPPR functionalization significantly enhanced the in vitro inhibitory effect of doxorubicin liposomes on glioblastoma cells and prolonged the median survival time of nude mice bearing intracranial glioblastoma. Finally, the immunofluorescence analysis evidenced that RGE-LS were able to penetrate through tumor vessels and stroma and deep into the whole tumor tissue. The results indicated that tumor-penetrating peptide functionalization is an effective strategy for enhancing the anti-glioblastoma effect of doxorubicin liposomes.

Novel biodegradable electrospun nanofibrous P(DLLA-CL) balloons for the treatment of vertebral compression fractures.

Significant interest has been expressed by the spinal surgeon community for the use of calcium phosphate cement (CPC) in the treatment of vertebral compression fractures (VCFs), but the water-induced collapsibility and poor mechanical properties limit its clinical use. Here we fabricated novel electrospun nanofibrous P(DLLA-CL) balloons (ENPBs) using the nanotechnique of electrospinning. The ENPBs could separate the cements from the surrounding environment, and therefore can prevent the water-induced collapsibility of CPC and eliminate cement leakage. The ENPBs filling with CPC had enough load-bearing ability to restore the height of the fractured vertebral body and had no obvious effects on the initial strength and stiffness of natural bones. Further, the ENPBs had good biodegradability and cell proliferation ability. Calcium can be released from ENPBs filling with CPC. All these results strongly demonstrate ENPBs can be potentially used as CPC filling containers that keep the advantages and eliminate the disadvantages of CPC. FROM THE CLINICAL EDITOR: Calcium phosphate cement (CPC) is a promising modality in vertebral compression fracture treatment, but its water-induced collapsibility limits clinical applications. This team of investigators fabricated novel nanofibrous balloons using electrospinning, which enabled the separation of CPC from its surrounding environment, and therefore prevented water-induced collapsibility of CPC and eliminated cement leakage while maintaining all the advantages of CPC treatment.

Recent progress in the application of atomic force microscopy for supported lipid bilayers.

In the past two decades, atomic force microscopy has been widely used for studying supported lipid bilayer related research, including the structure and dynamics of membranes and membrane proteins, and the interaction of membranes with chemical and biological molecules. The focus of this minireview is on the recent progress in the application of atomic force microscopy for supported lipid bilayers. Such progress mainly includes the application in the following aspects: submolecular-resolution imaging, in situ observation, and nanomechanics measurement.

Gadolinium-chitosan nanoparticles as a novel contrast agent for potential use in clinical bowel-targeted MRI: a feasibility study in healthy rats.

BACKGROUND: MRI is of increasing importance in the diagnostic evaluation of gastrointestinal diseases, with depiction of mucosal enhancement obtained with conventional intravenous contrast. Routine clinical use of contrast agents has been carried out using intravenous injection for mucosal imaging. Contrast agents that specifically target the intestinal mucosa are therefore needed to improve clinical imaging of the mucosal surface. PURPOSE: To synthesize a novel contrast agent for gadopentetic acid (Gd-DTPA)-loaded chitosan nanoparticles and observe the absorption of the nanoparticles in the colon wall of healthy rats by MR imaging in vivo. MATERIAL AND METHODS: A contrast agent was successfully synthesized by a modified emulsion coalescence method, and the resulting agents were characterized in detail by dynamic light-scattering spectroscopy and inductively coupled plasma emission spectroscopy. The cytotoxicity of Gd-chitosan nanoparticles was evaluated by an MTT assay. Gadolinium-chitosan (Gd@chitosan) nanoparticles were administered to the colon mucosa of healthy rats by rectal administration, and MRI scans in vivo were carried out with a 3.0 T imaging scanner at various time points. RESULTS: The prepared Gd@chitosan nanoparticles were ~420 nm in diameter with a 74.4% Gd-DTPA content. The MTT assay indicated little cytotoxicity. MRI results showed that nanoparticles can be retained in both the stratum submucosum and epithelial cells of the colon for almost 80 min. Transmission electron microscopy images further revealed that Gd@chitosan nanoparticles were localized inside the mucosal cells or intercellular space, while tissue from Gd-DTPA aqueous solution administration showed nothing. Due to the infusion of Gd@chitosan nanoparticles, the MR signal intensity of colon mucosa increased from about 6% to 35%, and the contrast enhancement was highest at 20 min after administration. CONCLUSION: Gd@chitosan nanoparticles with high Gd-DTPA content were successfully prepared for use as a novel MRI contrast agent. All results indicated that rectally administered Gd@chitosan nanoparticles have the potential for MRI diagnosis of colon mucosal disease.

Low Concentrations of Zinc Oxide Nanoparticles Cause Severe Cytotoxicity Through Increased Intracellular Reactive Oxygen Species.

With wide application of Zinc oxide (ZnO) nanoparticles, their biological toxicity has received more and more attention in recent years. In this research, two ZnO dispersions with different particle sizes, small size Zinc oxide (S-ZnO) and big size Zinc oxide (B-ZnO), were prepared using polycarboxylic acid as dispersant. We found that the S-ZnO nanoparticles showed stronger toxicity on Human Pulmonary Alveolar Epithelial Cells (HPAEpiC) under same concentration. Only 9 ppm S-ZnO could decrease HPAEpiC viability to about 50%, which means that, a small amount of well-dispersed ZnO nanoparticles in industrial production process may cause serious damage to the human body through oral inhalation. Focusing on mechanism for cytotoxicity, ZnO nanoparticles promoted generation and accumulation of Reactive Oxygen Species (ROS) in mitochondria via inhibiting Superoxide Dismutase (SOD) enzyme activity and reducing Glutathione (GSH) content. ROS in turn opened the mitochondrial Ca2+ pathway and lowered the Mitochondrial Membrane Potentials (MMP), leading to cell death. To simulate the lung environment in vitro, mixed dipalmitoyl phosphatidylcholine (DPPC) and ZnO nanoparticles (1:1) were incubated for 72 hours and then cytotoxicity was evaluated on HPAEpiC. Results showed that the cell viability was significantly increased, which proved that the DPPC effectively inhibited the toxicity of ZnO nanoparticles.