Self-Assembled ATP-Responsive DNA Nanohydrogel for Specifically Activated Fluorescence Imaging and Chemotherapy in Cancer Cells.

Tumor marker-responsive drug delivery systems have been developed for cancer imaging and chemotherapy. However, improving their ability of controlled drug release remains a challenge. In this study, we have developed an adenosine triphosphate (ATP)-responsive DNA nanohydrogel for specifically activated fluorescence imaging and chemotherapy in cancer cells. Acrylamide and acrydite-modified DNAs were polymerized to obtain DNA-grafted polyacrylamide copolymers. Then, the copolymers acted as the backbone of the nanohydrogel and were assembled by base complementation with ATP aptamer linkers to construct an ATP-responsive nanohydrogel. Meanwhile, the chemotherapeutic drug doxorubicin (DOX) was added and loaded into the ATP-responsive nanohydrogel during the assembly process. After endocytosis by cancer cells and response to a high intracellular ATP level, the DOX-loaded nanohydrogel disassembled due to the formation of aptamer/ATP complexes. Subsequently, the released DOX played a role in fluorescence imaging and chemotherapy of cancer cells. Through the ATP-responsive property and satisfying drug delivery capability, this nanohydrogel realized fluorescence imaging and specific cancer cell killing capabilities due to different intracellular ATP levels in normal and cancer cell lines. In summary, this study has provided a novel strategy of constructing a tumor microenvironment-responsive drug delivery system triggered by the tumor markers for tumor intracellular imaging and chemotherapy.

Biomimetic Design of Hollow Flower-Like g-C3N4@PDA Organic Framework Nanospheres for Realizing an Efficient Photoreactivity.

Organic framework polymers have attracted much interest due to the enormous potential design space offered by the atomically precise spatial assembly of organic molecular building blocks. The morphology control of organic frameworks is a complex issue that hinders the development of organic frameworks for practical applications. Biomimetic self-assembly is a promising approach for designing and fabricating multiple-functional nanoarchitectures. A bioinspired hollow flower-like organic framework nanosphere heterostructure comprised of carbon nitride and polydopamine (g-C3N4@PDA) is successfully synthesized via a mild and green method. This heterostructure can effectively avoid the agglomeration of nanosheets to better access the hollow nanospheres with high open-up specific surface area. The electron delocalization of g-C3N4 and PDA under visible light can largely promote photoelectron transfer and enhance the photocatalytic activity of the g-C3N4@PDA. Furthermore, the g-C3N4@PDA can effectively enhance the generation of reactive oxygen species under irradiation, which can lead to cell apoptosis and enhance the performance for cancer therapy. Therefore, the as-prepared g-C3N4@PDA provides a paradigm of highly efficient photocatalyst that can be used as nanomedicine toward cancer therapy. This study could open up a new avenue for exploiting more other potential hollow nanosphere organic frameworks.

Avertoxins A-D, Prenyl Asteltoxin Derivatives from Aspergillus versicolor Y10, an Endophytic Fungus of Huperzia serrata.

Aspergillus versicolor Y10 is an endophytic fungus isolated from Huperzia serrata, which showed inhibitory activity against acetylcholinesterase. An investigation of the chemical constituents of Y10 led to the isolation of four new prenylated asteltoxin derivatives, named avertoxins A-D (2-5), together with the known mycotoxin asteltoxin (1). In the present study, we report structure elucidation for 2-5 and the revised NMR assignments for asteltoxin and demonstrated that avertoxin B (3) is an active inhibitor against human acetylcholinesterase with the IC50 value of 14.9 µM (huperzine A as the positive control had an IC50 of 0.6 µM). In addition, the cytotoxicity of asteltoxin (1) and avertoxins A-D (2-5) against MDA-MB-231, HCT116, and HeLa cell lines was evaluated.

Switchable bifunctional stimuli-triggered poly-N-isopropylacrylamide/DNA hydrogels.

DNA-tethered poly-N-isopropylacrylamide copolymer chains, pNIPAM, that include nucleic acid tethers have been synthesized. They are capable of inducing pH-stimulated crosslinking of the chains by i-motif structures or to be bridged by Ag(+) ions to form duplexes. The solutions of pNIPAM chains undergo crosslinking at pH 5.2 or in the presence of Ag(+) ions to form hydrogels. The hydrogels reveal switchable hydrogel-to-solution transitions by the reversible crosslinking of the chains at pH 5.2 and the separation of the crosslinking units at pH 7.5, or by the Ag(+) ion-stimulated crosslinking of the chains and the reverse dissolution of the hydrogel by the cysteamine-induced elimination of the Ag(+) ions. The DNA-crosslinked hydrogels are thermosensitive and undergo reversible temperature-controlled hydrogel-to-solid transitions. The solid pNIPAM matrices are protected against the OH(-) or cysteamine-stimulated dissociation to the respective polymer solutions.

siRNA-Based Carrier-Free System for Synergistic Chemo/Chemodynamic/RNAi Therapy of Drug-Resistant Tumors.

Multiple drug-resistance mechanisms originate from defensive pathways in cancer and are associated with the unsatisfied efficacy of chemotherapy. The combination of small interfering RNA (siRNA) and chemotherapeutics provides a strategy for reducing drug efflux but requires more delivery options for clinical translation. Herein, multidrug resistance protein 1 (MDR1) siRNA is used as the skeleton to assemble chemotherapeutic cisplatin (CDDP) and divalent copper ion (Cu2+) for constructing a carrier-free Cu-siMDR-CDDP system. Cu-siMDR-CDDP specifically responds and disassembles in the acidic tumor microenvironment (TME). The released CDDP activates cascade bioreactions of NADPH oxidases and superoxide dismutase to generate hydrogen peroxide (H2O2). Then a Cu2+-catalyzed Fenton-like reaction transforms H2O2 to hydroxyl radicals (HO•) and causes glutathione (GSH) depletion to disrupt the redox adaptation mechanism of drug-resistant cancer cells. Besides, delivery of MDR1 siRNA is facilitated by HO•-triggered lysosome destruction, thus inhibiting P-glycoprotein (P-gp) expression and CDDP efflux. The unique design of Cu-siMDR-CDDP is to exploit siRNA as building blocks in regulating the self-assembly behavior, and integration of functional units simultaneously alleviates limitations caused by drug-resistance mechanisms. Such a carrier-free system shows synergistic chemo/chemodynamic/RNA interference therapy in suppressing tumor growth in vivo and has the reference value for overcoming drug resistance.

Multifunctional Carbon Monoxide Prodrug-Loaded Nanoplatforms for Effective Photoacoustic Imaging-Guided Photothermal/Gas Synergistic Therapy.

Multifunctional cancer treatments based on gas therapy combined with other cancer treatments have gained tremendous attention and hold great promise in biomedical applications. In this study, a carbon monoxide-releasing nanoplatform combined with near-infrared (NIR) laser-triggered photothermal therapy (PTT) was constructed. The nanoplatform was composed of manganese pentacarbonyl bromide (MnCO)-loaded g-carbon nitride/polypyrrole (CNPpy) nanomaterials (MnCO@CNPpy). MnCO can be triggered to produce CO under H2O2 conditions. Upon exogenous NIR light stimulation and tumor microenvironment-overexpressed H2O2, MnCO@CNPpy exhibited excellent CO generation performance and photothermal effect. The generation of CO induced intracellular oxidative stress and caused cell apoptosis. Additionally, photoacoustic (PA) imaging was performed to track the delivery and accumulation of the nanomaterial in tumor sites because of the great photothermal conversion of CNPpy. The presented MnCO@CNPpy nanoplatform displayed desirable PTT and CO therapy in the inhibition of tumor growth and may provide a promising strategy for multifunctional antitumor synergistic treatments.

Anti-washout carboxymethyl chitosan modified tricalcium silicate bone cement: preparation, mechanical properties and in vitro bioactivity.

Anti-washout CaF(2) stabilized C(3)S (F-C(3)S) bone cement was prepared by adding water-soluble carboxymethyl chitosan (CMCS) to the hydration liquid. The setting time, compressive strength and in vitro bioactivity of the CMCS modified F-C(3)S (CMCS-C(3)S) pastes were evaluated. The results indicate that CMCS-C(3)S pastes could be stable in the shaking simulated body fluid (SBF) after immediately mixed. The addition of CMCS significantly enhances the cohesion of particles, at the same time restrains the penetration of liquid, and thus endows the anti-washout ability. The setting times of the pastes increase with the increase of CMCS concentrations in the hydration liquid. Besides, the compressive strengths of CMCS-C(3)S pastes after setting for 1-28 days are lower than that of the pure F-C(3)S paste, but the sufficient strengths would be suitable for the clinical applications. The crystalline apatite deposited on the paste surface is retarded from 1 to 2 days for the addition of CMCS, but the quantities of deposited apatite are same after soaking in SBF for 3 days. As the result that pure C(3)S paste has shorter setting times than pure F-C(3)S paste, CMCS modified pure C(3)S pastes would have better anti-washout ability. Our study provides a convenient way to use C(3)S bone cement with excellent anti-washout ability when the pastes are exposed to biological fluids. The novel anti-washout CMCS-C(3)S bone cement with suitable setting times, sufficient strengths and in vitro bioactivity would have good prospects for medical application.

Simultaneous voltammetry detection of dopamine and uric acid in human serum and urine with a poly(procaterol hydrochloride) modified glassy carbon electrode.

In the present study, procaterol hydrochloride (ProH) was successfully electropolymerized onto a glass carbon electrode (GCE) with simply cyclic voltammetry scans to construct a poly(procaterol hydrochloride) (p-ProH) membrane modified electrode. Compared with the bare GCE, much higher oxidation peak current responses and better peak potentials separation could be obtained for the simultaneous oxidation of dopamine (DA) and uric acid (UA), owning to the excellent electrocatalytic ability of the p-ProH membrane. And it's based on that a square wave voltammetry (SWV) method was developed to selective and simultaneous measurement of DA and UA. Under the optimum conditions, the linear dependence of oxidation peak current on analyte concentrations were found to be 1.0-100 µmol/L and 2-100 µmol/L, giving detection limits of 0.3 µmol/L and 0.5 µmol/L for DA and UA, separately. The as prepared modified electrode shows simplicity in construction with the merits of good reproducibility, high stability, passable selectivity and nice sensitivity. Finally, the proposed p-ProH membrane modified electrode was successfully devoted to the detection of DA and UA in biological fluids such as human serum and urine with acceptable results.

A black phosphorus nanosheet-based siRNA delivery system for synergistic photothermal and gene therapy.

Gene therapy with small interfering RNA (siRNA) has been proved to be a promising technology to treat various diseases by hampering the production of target proteins. However, developing a delivery system that has high efficiency in transporting siRNA without obvious side effects remains a challenge. Herein, we designed a new survivin siRNA delivery system based on polyethyleneimine functionalized black phosphorus (BP) nanosheets which could suppress tumor growth by silencing survivin expression. Combined with the photothermal properties of the BP nanosheets, the presented delivery system shows excellent therapy efficiency for tumors. Therefore, the BP-based delivery system would be a promising tool for future clinical applications.

High photoluminescent carbon based dots with tunable emission color from orange to green.

A one-pot hydrothermal method was proposed for the synthesis of carbon based dots (CDs) with high quantum yield and controllable long-wavelength photoluminescence (PL). The PL mechanisms of the CDs were discussed, and a common model has been proposed. Furthermore, the obtained CDs showed excellent biocompatibility and high PLQYs (more than 20%), and presented great potential bio-applications.

Electrochemical investigation and determination of procaterol hydrochloride on poly(glutamic acid)/carboxyl functionalized multiwalled carbon nanotubes/polyvinyl alcohol modified glassy carbon electrode.

Poly(glutamic acid) (P-GLU)/carboxyl functionalized multiwalled carbon nanotubes (MWCNTs-COOH)/polyvinyl alcohol (PVA) modified glassy carbon electrode (GCE) has been successfully prepared and the electrochemical behavior of procaterol hydrochloride (ProH) was studied. The results show that the as-prepared modified electrode exhibits a good electrocatalytic property towards the oxidation of ProH in 0.2M phosphate buffer solution (PBS) (pH 6.0) due to the enhanced oxidation peak current at ~+0.59V. Under optimal reaction conditions, the oxidation peak current of ProH is proportional to its concentration in the linear dynamic ranges of 0.060 - 8.0µM (R = 0.9974), with a detection limit of 8.0 × 10-9M. Finally, this method was efficiently used for the determination of ProH in tablets and human urine with recoveries of 88.5~98.7% and 89.2 ~ 108.0%, respectively.

Intraperitoneal delivery of liposomal siRNA for therapy of advanced ovarian cancer.

PURPOSE: Intravenous (IV) delivery of siRNA incorporated into neutral liposomes allows efficient delivery to tumor tissue, and has therapeutic efficacy in preclinical proof-of-concept studies using EphA2-targeting siRNA. We sought to determine whether intraperitoneal (IP) delivery of these siRNA complexes was as effective at delivery and therapy as IV delivery. EXPERIMENTAL DESIGN: SiRNA was incorporated into the neutral liposome 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC). Alexa555-siRNA-DOPC was injected IP into nude mice bearing established ovarian tumors, and organs were collected for microscopic fluorescent examination. Subsequently, therapeutic efficacy of the IP versus IV routes was directly compared. RESULTS: Alexa555-siRNA in DOPC liposomes injected IP was diffusely distributed into intraperitoneal ovarian tumors. Delivery was also seen deeply into the liver and kidney parenchyma, suggesting that the predominant means of distribution was through the vasculature, rather than direct diffusion from the peritoneal cavity. In mice with orthotopic ovarian tumors, treatment with combined paclitaxel and IP EphA2-targeting siRNA-DOPC reduced tumor growth by 48-81% compared to paclitaxel/control siRNA-DOPC IP (HeyA8: 0.34 g v 0.66 g; SKOV3ip1: 0.04 v 0.21, p<0.01). This reduction was comparable to concurrently-treated mice with paclitaxel and EphA2 siRNA-DOPC injected IV, which showed a reduction in growth by 45-69% compared to paclitaxel/control siRNA-DOPC injected IV (HeyA8: 0.23g v. 0.42g; SKOV3ip1: 0.04 v. 0.13 g). CONCLUSIONS: IP injection of siRNA incorporated in DOPC allows intra-tumoral delivery and has therapeutic efficacy in orthotopic ovarian tumors. These findings may have therapeutic implications for siRNA-based strategies.

In vivo behaviors of Ca(OH)2 activated nano SiO2 (nCa/nSi=3) cement in rabbit model.

In vivo behaviors of Ca(OH)2 activated nano SiO2 (nCa/nSi=3, TCS) cement were investigated in the rabbit femoral defects using the poly(methyl methacrylate) (PMMA) as control. The deposited apatite and CaCO3 layers round TCS paste surfaces were completely used to construct the new bone tissue. TCS paste could stimulate the formation of new bone tissue in marrow tissue. The osteostimulation was mainly attributed to the proliferation and differentiation effects of Ca and Si ions released from TCS paste on the osteoprogenitor cells. However, Calcium-Silicate-Hydrate (C-S-H) gel in TCS paste was harder to degradate than Ca(OH)2. TCS paste kept the original shape during implantation, and could not provide the pores or spaces for further formation of bone tissue. Osteolytic defects induced by wear particles from TCS paste surface could not be completely avoided, because of the interfacial strain and the extensive micromotion between TCS paste surface and new bone tissue. Overall, our results indicated that Ca(OH)2 activated nano SiO2 cement was bioactivity and osteostimulation. The further improvements of Ca(OH)2 activated nano SiO2 cement should be done by achieving a balance between biological properties and mechanical performances.

pH-stimulated DNA hydrogels exhibiting shape-memory properties.

Nucleic acid-functionalized polyacrylamide chains that are cooperatively cross-linked by i-motif and nucleic acid duplex units yield, at pH 5.0, DNA hydrogels exhibiting shape-memory properties. Separation of the i-motif units at pH 8.0 dissolves the hydrogel into a quasi-liquid phase. The residual duplex units provide, however, a memory code in the quasi-liquid allowing the regeneration of the hydrogel shape at pH 5.0.

[Determination of free formaldehyde in cosmetics by pre-column derivatization, extraction inhibition and high performance liquid chromatography].

Pre-column derivatization and inhibition by solvent extraction were applied to determine free formaldehyde in cosmetics by high performance liquid chromatography (HPLC). Due to the rapid decomposition of formaldehyde donors in the derivatization, it is hard to detect the amount of the free formaldehyde in cosmetics. The formaldehyde directly reacted with 2,4-dinitrophenylhydrazine in acetonitrile-phosphate buffer (pH 2) (1:1, v/v) solution for 2 min, then dichloromethane extraction was used to induce the decomposition of formaldehyde donors. The extract was diluted with acetonitrile and then determined by HPLC. The formaldehyde derivative was separated on an Agilent C18 column (250 mm x 4.6 mm, 5 microm) at 30 degrees C with acetonitrile-water (60:40, v/v) as mobile phase at a flow rate of 1.0 mL/min, and detected at the wavelength of 355 nm. The recoveries were from 81% to 106% at the spiked levels of 50, 100, 500, 1 000 microg/g of formaldehyde in shampoo, milk, cream, hand cleaner, toothpaste, nail polish, powder separately, and the relative standard deviations (n = 6) were less than 5.0%. The limit of quantification of the formaldehyde in cosmetics was 50 microg/g. The method has been applied to the determination of free formaldehyde in real samples and the results showed that the release by formaldehyde donors was inhibited. The method has the advantages of simple operation, good accuracy and meets the requirement of determination of free formaldehyde in cosmetics.

Controlled synthesis of NaYF4 nanoparticles and upconversion properties of NaYF4:Yb, Er (Tm)/FC transparent nanocomposite thin films.

Monodisperse oleic acid stabilized pure NaYF(4) nanoparticles with controlled size and shape have been successfully synthesized by changing the initial reaction temperature. Transparent nanocomposite thin films consisting of NaYF(4):Yb, Er (Tm) upconverting nanoparticles (UCNPs) and fluorocarbon resin (FC) are deposited on the slide glass by dip-coating method. The results show that these nanocomposite thin films exhibit intense green and blue upconversion photoluminescence under 980 nm laser excitation and higher transparency than blank substrate. The NaYF(4):Yb,Er (Tm) nanoparticles and NaYF(4):Yb,Er (Tm)/FC nanocomposite thin films have been characterized by X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), scanning electron microscopy (SEM), SEM/back-scattered electron (BSE), atomic force microscopy (AFM), UV-Vis spectrophotometer (UVPC), and photoluminescence (PL) spectra. These nanocomposite thin films can be potentially used in solar cells field.

The reactivity of nano silica with calcium hydroxide.

The reactivity of nano silica (SiO2) with calcium hydroxide (Ca(OH)2) was evaluated and characterized in this study. Ca(OH)2 activated nano SiO2 takes place through an exothermic process, which is mainly attributed to the breakdown of Si-O-Si bonds. Ca(2+) offsets the charge imbalance and bonds to Si-OH and Si-O(-) giving rise to calcium silicate hydrate (CSH) gel. Care has to be taken that the reactivity of nano SiO2 with Ca(OH) 2significantly depends on the Q³ percentage in nano SiO 2. Q³ percentages significantly influence the reaction kinetic of nano SiO2 . The higher Q³ percentage results in a higher reaction degree of nano SiO2 with Ca(OH)2 and shorter setting times of the pastes. The higher Q³ percentage results in a lower total reaction heat of nano SiO2 with Ca(OH)2. It is suggested that the Q³ percentages of nano SiO2 should be in excess of 30% to keep the satisfactory setting properties of the pastes for the application requirements of bone cement.

Preparation and characterization of novel alkali-activated nano silica cements for biomedical application.

The major goal of this study was to investigate and characterize novel Ca(OH)(2)-activated nano silica (SiO(2)) cements for the potential application as a bone cement. Novel Ca(OH)(2)-activated nano-SiO(2) powders composed of Ca(OH)(2) and nano-SiO(2) were easily prepared, and the deionized water was used as the liquid phase. The initial and final setting times of Ca(OH)(2)-activated nano-SiO(2) cements with liquid to powder (L/P) ratios of 1.00-1.33 mL g(-1) range from 50 ± 1 to 120 ± 3 min and 96 ± 2 to 190 ± 5 min, respectively. The calorimetric curves indicate that the heat liberations of Ca(OH)(2)-activated nano-SiO(2) cements (105.57-138.01 J g(-1)) are lower than that of C(3)S (about 150 J g(-1)). The Ca(OH)(2)-activated nano-SiO(2) pastes are injectable under general extrusion force of 100 N, and their compressive strengths with suitable L/P ratios are 5-9 MPa, which is comparable with that of cancellous bone. The in vitro bioactivity of the Ca(OH)(2)-activated nano-SiO(2) cement pastes were investigated by soaking in simulated body fluid for various periods. The results show amorphous apatite deposits on the paste surfaces after soaking in simulated body fluid for 2 h. With the prolonged soaking time, amorphous apatite transforms into crystalline apatite after soaking for 1 day. The Ca(OH)(2)-activated nano-SiO(2) cements have short setting times, low heat liberation, injectability, suitable compressive strength, and excellent in vitro bioactivity, and may be used as bioactive bone cements for bone regeneration.

A novel sensitive detection platform for antitumor herbal drug aloe-emodin based on the graphene modified electrode.

This paper has presented a novel strategy to carry out direct and sensitive determination of antitumor herbal drug aloe-emodin in complex matrices based on the graphene-Nafion modified glassy carbon (GN/GC) electrode. This proposed modified electrode showed good electrochemical response towards aloe-emodin (AE). Compared with the multiwall carbon nanotubes (MWCNTs) modified electrode, the GN/GC electrode has the advantages of higher sensitivity and lower cost. Under the optimized conditions, the calibration curve for AE concentration was linear in the range from 5 nmol/L to 1 µmol/L with the detection limit of 2 nmol/L. In addition, the practical analytical performance of the GN/GC electrode was examined by evaluating the selective detection of AE in natural aloe extracts and human urine samples with satisfied recovery. Therefore, the GN/GC electrode may hold great promise for fast, simple and sensitive detection and biomedical analysis of AE in complex matrices.

Sustained small interfering RNA delivery by mesoporous silicon particles.

RNA interference (RNAi) is a powerful approach for silencing genes associated with a variety of pathologic conditions; however, in vivo RNAi delivery has remained a major challenge due to lack of safe, efficient, and sustained systemic delivery. Here, we report on a novel approach to overcome these limitations using a multistage vector composed of mesoporous silicon particles (stage 1 microparticles, S1MP) loaded with neutral nanoliposomes (dioleoyl phosphatidylcholine, DOPC) containing small interfering RNA (siRNA) targeted against the EphA2 oncoprotein, which is overexpressed in most cancers, including ovarian. Our delivery methods resulted in sustained EphA2 gene silencing for at least 3 weeks in two independent orthotopic mouse models of ovarian cancer following a single i.v. administration of S1MP loaded with EphA2-siRNA-DOPC. Furthermore, a single administration of S1MP loaded with-EphA2-siRNA-DOPC substantially reduced tumor burden, angiogenesis, and cell proliferation compared with a noncoding control siRNA alone (SKOV3ip1, 54%; HeyA8, 57%), with no significant changes in serum chemistries or in proinflammatory cytokines. In summary, we have provided the first in vivo therapeutic validation of a novel, multistage siRNA delivery system for sustained gene silencing with broad applicability to pathologies beyond ovarian neoplasms.