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Catalyzed oxygen insertion into C–H bonds represents a continuous challenge in chemistry. Particularly, driving this process at ambient temperature and aqueous media represents a “holy grail” in catalysis. We report on the catalyzed cascade transformations of l-tyrosine or l-phenylalanine to dopachrome in the presence of l-ascorbic acid/H2O2 as oxidizing mixture and CuFe-Prussian Blue-like nanoparticles, Fe3O4 nanoparticles or Au nanoparticles as catalysts. The process involves the primary transformation of l-tyrosine to l-DOPA that is further oxidized to dopachrome. The transformation of l-phenylalanine to dopachrome in the presence of CuFe-Prussian Blue-like nanoparticles and l-ascorbic acid/H2O2 involves in the first step the formation of l-tyrosine and, subsequently, the operation of the catalytic oxidation cascade of l-tyrosine to l-DOPA and dopachrome. Electron spin resonance experiments demonstrate that ascorbate radicals and hydroxyl radicals play cooperative functions in driving the different oxy...

A smart core-sheath nanofiber for non-adherent cell capture and release is demonstrated. The nanofibers are fabricated by single-spinneret electrospinning of poly(N-isopropylacrylamide) (PNIPAAm), polycaprolactone (PCL) and nattokinase (NK) solution blends. The self-assembly of PNIPAAm and PCL blends during the electrospinning generates the core-sheath PCL/PNIPAAm nanofibers with PNIPAAm as the sheath. The PNIPAAm-based core-sheath nanofibers are switchable between hydrophobicity and hydrophilicity with temperature change and enhance stability in the blood. When the nanofibers come in contact with blood, the NK is released from the nanofibers to resist platelet adhesion on the nanofiber surface, facilitating the direct capture and isolation of red blood cells (RBCs) from the blood above phase-transition temperature of PNIPAAm. Meanwhile, the captured RBCs are readily released from the nanofibers with temperature stimuli in an undamaged manner. The release efficiency of up to 100% is obtained while maintaining cellular integrity and function. This work presents promising nanofibers to effectively capture non-adherent cells and release for subsequent molecular analysis and diagnosis of single cells.

A newly glycopolymer-patterned surface for capturing red blood cells (RBCs) is demonstrated. Our strategy is based on the surface-initiated photopolymerization of 2-acryl-amido-2-methylpropane sulfonic acid (AMPS) on a thermoplastic elastomer, the patterning of poly(d-gluconamidoethyl methacrylate) (PGAMA, glycopolymer) micro-domains on the PAMPS layer with photomask-assisted photolithography, followed by the generation of a phytohemagglutinin (PHA) array on the patterned surface through lectin-carbohydrate recognition. We demonstrate that the bi-component polymer-patterned surface with high lateral resolution is successfully fabricated; the PAMPS layer with patterned glycopolymer domains remains hydrophilic to resist non-specific plasma protein adsorption and cell adhesion; the PHA array on the patterned PGAMA domains induces nearly no platelet adhesion on the patterned surface, but shows high capability for capturing RBCs in the blood, and in addition, the captured RBCs maintain cellular integrity and function. Our work presented herein not only paves a new way for capturing RBCs from the blood, but also establishes a basic principle to capture non-adherent cells in the blood or biological fluid without damage.

Although bio-inspired polydopamine (PDA) micro/nano-particles show great promise for biomedical applications, the knowledge on the interactions between micro/nano-particles and platelets is still lacking. Here, we fabricate PDA-coated micro/nano-particles and investigate the platelet–particle surface interactions. Our strategy takes the advantage of facile PDA coating on polystyrene (PS) microsphere to fabricate particles with varied sizes and surface properties, and the chemical reactivity of PDA layers to immobilize fibrinogen and bovine serum albumin to manipulate platelet activation and adhesion. We demonstrate that PS particles activate the platelets in the size-dependent manner, but PDA nanoparticles have slight effect on platelet activation; PS particles promote platelet adhesion while PDA particles reduce platelet adhesion on the patterned surface; Particles interact with platelets through activating the glycoprotein integrin receptor of platelets and providing physical sites for initial platelet adhesion. Our work sheds new light on the interaction between platelets and particles, which provides the basic principle to select biocompatible micro/nano-particles in biomedical field.

A novel hydrophilic PAMPS-PAAm brush pattern is fabricated to selectively capture blood cells from whole blood. PAMPS brushes provide antifouling surfaces to resist protein and cell adhesion while PAAm brushes effectively entrap targeted proteins for site-specific and cell-type dependent capture of blood cells.

Detection of dysfunctional and apoptotic cells plays an important role in clinical diagnosis and therapy. To develop a portable and user-friendly platform for dysfunctional and aging cell detection, we present a facile method to construct 3D patterns on the surface of styrene-b-(ethylene-co-butylene)-b-styrene elastomer (SEBS) with poly(ethylene glycol) brushes. Normal red blood cells (RBCs) and lysed RBCs (dysfunctional cells) are used as model cells. The strategy is based on the fact that poly(ethylene glycol) brushes tend to interact with phosphatidylserine, which is in the inner leaflet of normal cell membranes but becomes exposed in abnormal or apoptotic cell membranes. We demonstrate that varied patterned surfaces can be obtained by selectively patterning atom transfer radical polymerization (ATRP) initiators on the SEBS surface via an aqueous-based method and growing PEG brushes through surface-initiated atom transfer radical polymerization. The relatively high initiator density and polymerization temperature facilitate formation of PEG brushes in high density, which gives brushes worm-like morphology and superhydrophilic property; the tendency of dysfunctional cells adhered on the patterned surfaces is completely different from well-defined arrays of normal cells on the patterned surfaces, providing a facile method to detect dysfunctional cells effectively. The PEG-patterned surfaces are also applicable to detect apoptotic HeLa cells. The simplicity and easy handling of the described technique shows the potential application in microdiagnostic devices.

Hemolysis of red blood cells (RBCs) caused by implant devices in vivo and nonpolyvinyl chloride containers for RBC preservation in vitro has recently gained much attention. To develop blood-contacting biomaterials with long-term antihemolysis capability, we present a facile method to construct a hydrophilic, 3D hierarchical architecture on the surface of styrene-b-(ethylene-co-butylene)-b-styrene elastomer (SEBS) with poly(ethylene oxide) (PEO)/lecithin nano/microfibers. The strategy is based on electrospinning of PEO/lecithin fibers onto the surface of poly [poly(ethylene glycol) methyl ether methacrylate] [P(PEGMEMA)]-modified SEBS, which renders SEBS suitable for RBC storage in vitro. We demonstrate that the constructed 3D architecture is composed of hydrophilic micro- and nanofibers, which transforms to hydrogel networks immediately in blood; the controlled release of lecithin is achieved by gradual dissolution of PEO/lecithin hydrogels, and the interaction of lecithin with RBCs maintains the membrane flexibility and normal RBC shape. Thus, the blood-contacting surface reduces both mechanical and oxidative damage to RBC membranes, resulting in low hemolysis of preserved RBCs. This work not only paves new way to fabricate high hemocompatible biomaterials for RBC storage in vitro, but provides basic principles to design and develop antihemolysis biomaterials for implantation in vivo.

Combination chemotherapy with time-programmed administration of multiple drugs is a promising method for cancer treatment. However, realizing time-programmed release of combined drugs from a single carrier is still a great challenge in enhanced cancer therapy. Here, an implantable trilayer structured fiber device is developed to achieve time-programmed release of combined drugs for synergistic treatment of breast cancer. The fiber device is prepared by a modified microfluidic-electrospinning technique. The glycerol solution containing chemotherapy agent doxorubicin (Dox) forms the internal periodic cavities of the fiber, and poly(l-lactic acid) and poly(ε-caprolactone) containing the angiogenesis inhibitor apatinib (Apa) form the double walls of the fiber. Rapid release of Dox can be obtained by adjusting the wall thickness of the cavities, meanwhile sustained release of Apa is achieved through the slow degradation of the fiber matrix. After the fiber device is implanted subcutaneously near to the implanted solid tumor of mice, an excellent synergistic therapeutic effect is achieved through time-programmed release of the combined dual drugs. The fiber device provides a platform to sequentially co-deliver dual or multiple drugs for enhanced combined therapeutic efficacy.

Platelets have exhibited capabilities beyond clotting in recent years. Most of their functions are related to the nature of platelet adhesion. Establishing a facile method to understand the platelet adhesion and assess the platelet function through the mechanism and mechanics of adhesion is highly desired. Here, we report a generally applicable UV lithography technique with a photomask, which performs selective surface functionalization on large substrate areas, for creating stable, physical adhesive sites in the range of 12 μm to 3 μm. Our study demonstrated that the patterned surface facilitated probing of single platelet adhesion in a quantitative manner, and rendered platelets sensitive to adhesive proteins even at a low protein concentration. In addition, the platelet function in the presence of antiplatelet (anticancer) agents on platelets could be accurately estimated based on single platelet adhesion (SPA). This work paves a new way to understand and assess the blood platelet function. The SPA assay methodology has the potential to enable a rapid, accurate point-of-care platform suitable for evaluation of platelet function, detection of dysfunctional platelets, and assay of drug effects on platelets in cancer patients.

Contrary to a prevailing concept on protein adsorption and cell adhesion, novel micropatterned polyacrylamide (PAAm) brushes that can resist cell adhesion but promote protein retention are created through patterning of ATRP initiators and surface-initiated ATRP on a polymer substrate.

Platelets play a fundamental role in thrombus formation and in the pathogenesis of arterial thrombosis. Patterning surfaces for controlled platelet adhesion paves the way for adhesion and activation mechanisms in platelets and detection of platelet functional defects. Here, a new and simple method based on controlled polymerization of 2-methacryloyloxyethyl phosphorylcholine (MPC) on the surface of styrene-block-(ethylene-co-butylene)-block-styrene (SEBS) is shown. The competition between polymerization and degradation enables platelet adhesion on SEBS to be switched on and off. The adhesive sites of the platelets can be down to single cell level, and the dysfunctional platelets can be quantitatively detected.

Manipulation of cell–cell interactions has potential applications in basic research and cell-based therapy. Herein, using a combination of metabolic glycan labelling and bio-orthogonal click reaction, we engineer cell membranes with β-cyclodextrin and subsequently manipulate cell behaviours via photo-responsive host-guest recognition. With this methodology, we demonstrate reversible manipulation of cell assembly and disassembly. The method enables light-controllable reversible assembly of cell–cell adhesion, in contrast with previously reported irreversible effects, in which altered structure could not be reused. We also illustrate the utility of the method by designing a cell-based therapy. Peripheral blood mononuclear cells modified with aptamer are effectively redirected towards target cells, resulting in enhanced cell apoptosis. Our approach allows precise control of reversible cell–cell interactions and we expect that it will promote further developments of cell-based therapy., Reversible manipulation of cell-cell interactions has potential applications in basic research and cell-based therapy. Here the authors control cell-cell adhesion in vitro with light, by modifying the surface sugars of cells to display β-cyclodextrin, which recognises one isoform of light-isomerizable azobenzene linkers.

A versatile platform allowing capture and detection of normal and dysfunctional cells on the same patterned surface is important for accessing the cellular mechanism, developing diagnostic assays, and implementing therapy. Here, an original and effective method for fabricating binary polymer brushes pattern is developed for controlled cell adhesion. The binary polymer brushes pattern, composed of poly(N-isopropylacrylamide) (PNIPAAm) and poly[poly(ethylene glycol) methyl ether methacrylate] (POEGMA) chains, is simply obtained via a combination of surface-initiated photopolymerization and surface-activated free radical polymerization. This method is unique in that it does not utilize any protecting groups or procedures of backfilling with immobilized initiator. It is demonstrated that the precise and well-defined binary polymer patterns with high resolution are fabricated using this facile method. PNIPAAm chains capture and release cells by thermoresponsiveness, while POEGMA chains possess high capability to capture dysfunctional cells specifically, inducing a switch of normal red blood cells (RBCs) arrays to hemolytic RBCs arrays on the pattern with temperature. This novel platform composed of binary polymer brush pattern is smart and versatile, which opens up pathways to potential applications as microsensors, biochips, and bioassays.

Janus kinase (JAK) 2 plays a pivotal role in the tumorigenesis of signal transducers and activators of transcription (STAT) 3 constitutively activated solid tumors. JAK2 mutations are involved in the pathogenesis of various types of hematopoietic disorders, such as myeloproliferative disorders, polycythemia vera, essential thrombocythemia, and primary myelofibrosis. Thus, small-molecular inhibitors targeting JAK2 are potent for therapy of these diseases. In this study, we screened 1,062,608 drug-like molecules from the ZINC database and 2080 natural product chemicals. We identified a novel JAK family kinase inhibitor, dehydrocrenatidine, that inhibits JAK-STAT3-dependent DU145 and MDA-MB-468 cell survival and induces cell apoptosis. Dehydrocrenatidine represses constitutively activated JAK2 and STAT3, as well as interleukin-6-, interferon-α-, and interferon-γ-stimulated JAK activity, and STAT phosphorylation, and suppresses STAT3 and STAT1 downstream gene expression. Dehydrocrenatidine inhibits JAKs-JH1 domain overexpression-induced STAT3 and STAT1 phosphorylation. In addition, dehydrocrenatidine inhibits JAK2-JH1 kinase activity in vitro. Importantly, dehydrocrenatidine does not show significant effect on Src overexpression and epidermal growth factor-induced STAT3 activation. Our results indicate that dehydrocrenatidine is a JAK-specific inhibitor.

a b s t r a c t There is an urgent need to develop blood-contacting biomaterials with long-term anti-hemolytic capabil- ity. To obtain such biomaterials, we coaxially electrospin (ascorbic acid (AA) and lecithin)/poly (ethylene oxide) (PEO) core-shell nanofibers onto the surface of styrene-b-(ethylene-co-butylene)-b-styrene elas- tomer (SEBS) that has been grafted with poly (ethylene glycol) (PEG) chains. Our strategy is based on that the grafted layers of PEG render the surface hydrophilic to reduce the mechanical injure to red blood cells (RBCs) while the AA and lecithin released from nanofibers on blood-contacting surface can actively interact with RBCs to decrease the oxidative damage to RBCs. We demonstrate that (AA and lecithin)/PEO core-shell structured nanofibers have been fabricated on the PEG grafted surface. The binary release of AA and lecithin in the distilled water is in a controlled manner and lasts for almost 5 days; during RBCs preservation, AA acts as an antioxidant and lecithin as a lipid supplier to the membrane of erythrocytes, resulting in low mechanical fragility and hemolysis of RBCs, as well as high deformability of stored RBCs. Our work thus makes a new approach to fabricate blood-contacting biomaterials with the capability of long-term anti-hemolysis.

Epidermal growth factor receptor (EGFR) is a clinical therapeutic target to treat a subset of non–small cell lung cancer (NSCLC) harboring EGFR mutants. However, some patients with a similar kind of EGFR mutation show intrinsic resistance to tyrosine kinase inhibitors (TKI). It indicates that other key molecules are involved in the survival of these cancer cells. We showed here that 2-[(aminocarbonyl)amino]-5 -(4-fluorophenyl)-3- thiophenecarboxamide (TPCA-1), a previously reported inhibitor of IκB kinases (IKK), blocked STAT3 recruitment to upstream kinases by docking into SH2 domain of STAT3 and attenuated STAT3 activity induced by cytokines and cytoplasmic tyrosine kinases. TPCA-1 is an effective inhibitor of STAT3 phosphorylation, DNA binding, and transactivation in vivo. It selectively repressed proliferation of NSCLC cells with constitutive STAT3 activation. In addition, using pharmacologic and genetic approaches, we found that both NF-κB and STAT3 could regulate the transcripts of interleukin (IL)-6 and COX-2 in NSCLC harboring EGFR mutations. Moreover, gefitinib treatment only did not efficiently suppress NF-κB and STAT3 activity. In contrast, we found that treatment with TKIs increased phosho-STAT3 level in target cells. Inhibiting EGFR, STAT3, and NF-κB by combination of TKIs with TPCA-1 showed increased sensitivity and enhanced apoptosis induced by gefitinib. Collectively, in this work, we identified TPCA-1 as a direct dual inhibitor for both IKKs and STAT3, whereas treatment targeting EGFR only could not sufficiently repress NF-κB and STAT3 pathways for lung cancers harboring mutant EGFR. Therefore, synergistic treatment of TPCA-1 with TKIs has potential to be a more effective strategy for cancers. Mol Cancer Ther; 13(3); 617–29. ©2014 AACR.

Surface-initiated atom transfer radical polymerization (SI-ATRP) is a versatile tool for surface functionalization in a well-controlled manner. However, surface modification of styrenic thermoplastic elastomers (STPEs) faces a great challenge because immobilization of typical ATRP initiators onto STPEs needs to be carried out in organic solvent, which dissolves and destroys the STPEs film. In this work, a simple aqueous-based route is developed to immobilize ATRP initiators, Br, onto the surface of styrene-b-(ethylene-co-butylene)-b-styrene elastomer (SEBS), chosen as a model copolymer of STPEs. In such a way, functional polymer brushes of ethylene glycol methyl ether methacrylate (OEGMA) are successfully prepared from the surface of SEBS. Kinetic investigations show an approximately linear relationship between grafting density and reaction time, indicating the growth of chains is coincident with a "controlled" process. C-Br bonds directly connected to benzene rings on the SEBS-Br surfaces are demonstrated to be effective initiation sites for SI-ATRP. The even coverage of the surface by well-defined P(OEGMA) brushes enable SEBS films to exhibit excellent resistance to protein adsorption and platelet adhesion as well as low hemolysis ratio. This work not only manipulates the SEBS surface to substantially improve its bio-compatibility, but paves a way to facilitate SI-ATRP on the surface of styrene-based block copolymers (SBCs). (C) 2013 Published by Elsevier B.V.

Platelets play a fundamental role in thrombus formation and in the pathogenesis of arterial thrombosis. Patterning surfaces for controlled platelet adhesion paves the way for adhesion and activation mechanisms in platelets and detection of platelet functional defects. Here, a new and simple method based on controlled polymerization of 2-methacryloyloxyethyl phosphorylcholine (MPC) on the surface of styrene-block-(ethylene-co-butylene)-block-styrene (SEBS) is shown. The competition between polymerization and degradation enables platelet adhesion on SEBS to be switched on and off. The adhesive sites of the platelets can be down to single cell level, and the dysfunctional platelets can be quantitatively detected.

A newly developed nanofiber platform for nonadherent cell capture and release is demonstrated. The nanofiber platform is fabricated by single-spinneret electrospinning of bovine serum albumin (BSA)-conjugated poly(N-isopropylacrylamide) (PNIPAAm) and polycaprolactone blends. The nanofibers possess core-sheath structure with PNIPAAm as the sheath, which render the nanofibers switchable between hydrophilicity and hydrophobicity with the temperature change. As a result of BSA immobilization on the sheath part, the nanofibers resist platelet adhesion in the blood, facilitating the direct capture and isolation of red blood cells (RBCs) from the blood. Meanwhile, the captured RBCs are readily released from the nanofibers with the temperature stimuli. The capture and release efficiencies of up to 100% are achieved while maintaining cellular integrity and function. This work presents a promising platform to capture and release nonadherent cell effectively for subsequent molecular analysis and disease diagnosis.

Yanjie Yang,1,2 Jing Nan,3 Jianwen Hou,3 Bianfei Yu,1 Tong Zhao,1 Shuang Xu,1 Shuangyu Lv,2 Haixia Zhang1 1Key Laboratory of Nonferrous Metal Chemistry and Resources Utilization of Gansu Province, Lanzhou University, Lanzhou, 2School of Medicine, Henan University, Kaifeng, 3School of Life Sciences, Lanzhou University, Lanzhou, People’s Republic of China Abstract: In this study, we synthesized water-soluble fluorescent gold nanoclusters (Au NCs) stabilized with dihydrolipoic acid (DHLA). The cytotoxicity of these Au NCs was then assessed in the normal human hepatic cell line (L02) and the human hepatoma cell line (HepG2) at different exposure times. Cell viability was normal in both cell lines at 24hours and 48hours; however, the growth of HepG2 cells was significantly inhibited at 72hours. The change in lactate dehydrogenase level was strongly correlated with cell viability after 72hours incubation with DHLA–capped Au NCs, and the increase in cellular reactive oxygen species may be related to the decrease in cell viability. Growth inhibition of HepG2 cells was possibly due to difficultly passing the checkpoint between G1 phase and S phase. The anticancer activity of DHLA–capped Au NCs should be considered when used in biomedical imaging and drug delivery. Keywords: gold nanocluster, cytotoxicity, cell viability, reactive oxygen species, cell cycle