Thermal and Reactive Oxygen Species Dual-Responsive OEGylated Polysulfides with Oxidation-Tunable Lower Critical Solution Temperatures.

In this work, two monomethoxy oligo(ethylene glycol) (OEG)-substituted episulfides are prepared and a series of polysulfides are synthesized with subsequent ring-opening polymerization. The OEGylated polysulfides exhibit thermal and reactive oxygen species (ROS) dual-responsive behavior. Their lower critical solution temperatures (LCSTs) are close to human body temperature and depend on the degree of polymerization and OEG length. Notably, the LCST of the polysulfide increases linearly with the oxidation degree by H2 O2 , showing a highly tunable change regulated by the ratio between hydrophobic sulfide and hydrophilic sulfoxide/sulfone in the backbone. Further, the OEGylated polysulfide can act as a ROS scavenger to protect red blood cells (RBCs) from oxidative damage in an RBCs aging model in vitro. This work paves a facile way to synthesize LCST-tunable polysulfides, which hold great promise in biological applications.

Hydrogen Peroxide and Glutathione Dual Redox-Responsive Nanoparticles for Controlled DOX Release.

Polymer nanoparticulate drug delivery systems that respond to reactive oxygen species (ROS) and glutathione (GSH) simultaneously at biologically relevant levels hold great promise to improve the therapeutic efficacy to cancer cells with reduced side effects of chemo drugs. Herein, a novel redox dual-responsive amphiphilic block copolymer (ABP) that consists of a hydrophilic poly (ethylene oxide) block and a hydrophobic block bearing disulfide linked phenylboronic ester group as pendant is synthesized, and the DOX loaded nanoparticles (BSN-DOX) based on ABPs with varied hydrophobic block length are fabricated for DOX delivery. The self-immolative leaving reaction of phenylboronic ester triggered by extracellular ROS and the cleavage of disulfide linkages induced by intracellular GSH both lead to rapid DOX release from BSN-DOX, resulting in an on-demand DOX release. Moreover, BSN-DOX show better tumor inhibition and lower side effects in vivo compared with free drug.

Inhibition of Inflammation-Associated Thrombosis with ROS-Responsive Heparin-DOCA/PVAX Nanoparticles.

Inflammation-associated thrombosis is a non-negligible source of mortalities and morbidities worldwide. To manipulate inflammation-associated coagulation, nanoparticles that contain anti-inflammatory polymer (copolyoxalate containing vanillyl alcohol, PVAX) and anti-thrombotic heparin derivative deoxycholic acid (Hep-DOCA) are prepared. The strategy takes advantage of the reducted side effects of heparin through heparin conjugation, achievement of long-term anti-inflammation by inflammation-trigged release of anti-inflammatory agents, and formation of PVAX/heparin-DOCA nanoparticles by co-self-assembly. It is demonstrated that the Hep-DOCA conjugate and PVAX are synthesized successfully; PVAX and Hep-DOCA nanodrugs (HDP) are obtained by co-assembly; the HDP nanoparticles effectively reduce the inflammation and coagulation without inducing lethal bleeding both in vivo and in vitro. The method provided here is versatile and effective, which paves new way to develop nanodrugs to treat inflammation-associated thrombosis safely.

Construction of K+ responsive surface on SEBS to reduce the hemolysis of preserved erythrocytes.

Hemolysis of stored erythrocytes is a big obstacle for the development of new plasticizer-free polymer containers. Hemolysis is mainly caused by cell membrane oxidation and cation leaks from the intracellular fluid during storage. To construct an anti-hemolytic surface for a plasticizer-free polymer, we fabricated 2-O-α-d-glucopyranosyl-l-ascorbic acid (AA-2G)-loaded polycaprolactone (PCL)-crown ether micro/nanofibers on the surface of styrene-b-(ethylene-co-butylene)-b-styrene (SEBS). Our strategy is based on the sensitive response of the crown ether to leaked potassium, causing the release of AA-2G, the AA-2G can then remove the excess ROS, maintaining the Na/K-pump activity and the cell integrity. We demonstrated that the PCL-crown ether micro/nanofibers have been well prepared on the surface of SEBS; the micro/nanofibers provide a sensitive response to excess K+ and trigger the rapid release of AA-2G. AA-2G then acts as an antioxidant to reduce the excess ROS and maintain the Na/K-pump activity to mitigate cation leaks, resulting in the reduced hemolysis of the preserved erythrocytes. Our work thus provides a novel method for the development of plasticizer-free polymers for the storage of erythrocytes, and has the potential to be used to fabricate long-term anti-hemolytic biomaterials for in vivo use.

Multiple microarrays of non-adherent cells on a single 3D stimuli-responsive binary polymer-brush pattern.

We have developed a 3D smart binary polymer-brush pattern on the polymer substrate for inducing multiple cell microarrays aided by a lectin and temperature. The binary polymer-brush pattern composed of poly(N-isopropylacrylamide) (PNIPAM) and poly(d-gluconamidoethyl methacrylate) (PGAMA) brushes is fabricated by combining the photolithography technique with a surface-initiated photo-polymerization (SIPP) method. We demonstrate that well-defined binary polymer-brush patterns with high resolution are fabricated using this facile method. The patterned hierarchical PNIPAM brushes exhibit reversible switching to the adhesion of red blood cells (RBCs) induced by thermo-responsiveness. The PGAMA brush domains resist adhesion of RBCs but bind specifically with concanavalin A (Con A), forming a lectin pattern to capture RBCs in a site-specific manner. Therefore, multiple cell microarrays on a single platform are generated with the aid of Con A and temperature. This novel platform composed of a smart binary polymer-brush pattern is versatile and specific, which opens up pathways to potential applications such as microsensors, biochips and bioassays.

Bioinspired Antioxidant Defense System Constructed by Antioxidants-Eluting Electrospun F127-Based Fibers.

Cells were continuously exposed to oxidative damage by overproduction of reactive oxygen species (ROS) when they contacted implanted biomaterials. The strategy to prevent cells from oxidative injures remains a challenge. Inspired by the antioxidant defense system of cells, we constructed a biocompatible and ROS-responsive architecture on the substrate of styrene-b-(ethylene-co-butylene)-b-styrene elastomer (SEBS). The strategy was based on fabrication of architectures through reactive electrospinning of mixture including SEBS, acylated Pluronic F127, copolymer of poly(ethylene glycol) diacrylate and 1,2-ethanedithiol (PEGDA-EDT), and antioxidants (AA-2G) and ROS-triggered release of AA-2G from microfibers to detoxify the excess ROS. We demonstrated that the stable and hydrophilic architecture was constructed by phase separation of SEBS/F127 components and cross-linking between polymer chains during electrospinning; the ROS-responsive fibers controlled the release of AA-2G and the interaction of AA-2G with ROS reduced the oxidative damage to cells. The bioinspired architecture not only reduced mechanical and oxidative damage to cells but also maintained normal ROS level for physiological hemostasis. This work provides basic principles to design and develop antioxidative biomaterials for implantation in vivo.

Measuring Interfacial Adhesion of Carbon Nanotube Bundles and Electrospun Polymer Fibers.

Adhesion at the dissimilar interface of single wall carbon nanotube (SWCNT) bundles and electrospun polymer nanofibers (Espun) was directly measured using a nano cheese cutter. A sample fiber suspended at the tip of an atomic force microscope cantilever and another freestanding fiber on a mica substrate were arranged in a cross-cylinder configuration and allowed to adhere. External tensile load led to spontaneous detachment or "pull-off". Adhesion was shown to be dominated by van der Waals attraction, and the detachment process conforms to the classical Maugis parameter in the JKR-DMT transition.

Cell-inspired biointerfaces constructed from patterned smart hydrogels for immunoassays in whole blood.

Immunoassays have shown great advances in the fields of biomedical diagnosis. However, successful immunoassays in blood plasma or whole blood based on the designed biointerfaces are still rare. Here, a newly cell-inspired biointerface for immunoassays in blood is demonstrated. Inspired by the high resistance to protein and cell adhesion and extraordinary biological recognition of stem cells, the biointerfaces are constructed by patterning smart hydrogels (PNIPAAm-co-PNaAc) on hydrophilic layers (PEG), followed by immobilization of antibodies on the patterned hydrogels. The hierarchical biointerfaces are hydrophilic to resist blood plasma and blood cell adhesion, but exhibit high affinity to the target antigens. As a result, successful immunoassays in blood are achieved. In addition, the detection signal is further enhanced by the manipulation of the phase transition of the smart hydrogels with temperature, and the sensitivity is higher than that of the widely-used poly(acrylic acid)/(polyacrylate) platform. The biointerface is versatile and effective in antibody-antigen recognition, which offers a potential new approach for developing highly sensitive immunoassays in blood.

Capturing red blood cells from the blood by lectin recognition on a glycopolymer-patterned surface.

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.

A novel hydrophilic polymer-brush pattern for site-specific capture of blood cells from whole blood.

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.

Binary release of ascorbic acid and lecithin from core-shell nanofibers on blood-contacting surface for reducing long-term hemolysis of erythrocyte.

There is an urgent need to develop blood-contacting biomaterials with long-term anti-hemolytic capability. 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 elastomer (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.

Construction of 3D micropatterned surfaces with wormlike and superhydrophilic PEG brushes to detect dysfunctional 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.

Effect of surface interactions on adhesion of electrospun meshes on substrates.

Despite the importance of adhesion between electrospun meshes and substrates, the knowledge on adhesion mechanism and the method to improve the adhesion remain limited. Here, we precisely design the model system based on electrospun poly(ethylene oxide) (PEO) meshes and the substrate of styrene-b-(ethylene-co-butylene)-b-styrene elastomer (SEBS), and quantitatively measure the adhesion with a weight method. The surfaces of SEBS with different roughness are obtained by casting SEBS solution on the smooth and rough glass slides, respectively. Then, the surfaces of casted SEBS are respectively grafted with PEG oligomers and long PEG chains much larger than the entanglement molecular weight by surface-initiated atom transfer radical polymerization (SI-ATRP) of poly(ethylene glycol) methyl ether methacrylate (PEGMA). The detached surfaces of SEBS and electrospun fibers after adhesion measurements are analyzed by scanning electron microscopy (SEM). The adhesive force and adhesion energy are found to lie in the range from 68 to 220 mN and from 12 to 46 mJ/m(2), respectively, which are slightly affected by surface roughness of substrate but mainly determined by surface interactions. Just as the chemical cross-linking induces the strong adhesion, the chain entanglements on the interface lead to the higher adhesion than those generated by hydrophilic-hydrophobic interactions and hydrophilic interactions. The long grafted chains and the enhanced temperature facilitate the chain entanglements, resulting in the strong adhesive force. This work sheds new light on the adhesion mechanism at molecular level, which may be helpful to improve the adhesion between the electrospun fibers and substrates in an environmentally friendly manner.

Shear adhesion strength of aligned electrospun nanofibers.

Inspiration from nature such as insects' foot hairs motivates scientists to fabricate nanoscale cylindrical solids that allow tens of millions of contact points per unit area with material substrates. In this paper, we present a simple yet robust method for fabricating directionally sensitive shear adhesive laminates. By using aligned electrospun nylon-6, we create dry adhesives, as a succession of our previous work on measuring adhesion energies between two single free-standing electrospun polymer fibers in cross-cylinder geometry, randomly oriented membranes and substrate, and peel forces between aligned fibers and substrate. The synthetic aligned cylindrical solids in this study are electrically insulating and show a maximal Mode II shear adhesion strength of 27 N/cm(2) on a glass slide. This measured value, for the purpose of comparison, is 270% of that reported from gecko feet. The Mode II shear adhesion strength, based on a commonly known "dead-weight" test, is 97-fold greater than the Mode I (normal) adhesion strength of the same. The data indicate a strong shear binding on and easy normal lifting off. Anisotropic adhesion (Mode II/Mode I) is pronounced. The size and surface boundary effects, crystallinity, and bending stiffness of fibers are used to understand these electrospun nanofibers, which vastly differ from otherwise known adhesive technologies. The anisotropic strength distribution is attributed to a decreasing fiber diameter and an optimized laminate thickness, which, in turn, influences the bending stiffness and solid-state "wettability" of points of contact between nanofibers and surface asperities.

Controlled lecithin release from a hierarchical architecture on blood-contacting surface to reduce hemolysis of stored red blood cells.

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.

Superhydrophobic coating of elastomer on different substrates using a liquid template to construct a biocompatible and antibacterial surface.

The construction of biocompatible and antibacterial surfaces is becoming increasingly important. However, most of the existing techniques require multi-step procedures, stringent conditions and specific substrates. We present here a facile method to create a biocompatible and antibacterial surface on virtually any substrate under ambient conditions. The strategy is based on casting a highly adherent elastomer, styrene-b-(ethylene-co-butylene)-b-styrene, from a solvent mixture of xylene and decanol, in which decanol acts as both a polymer precipitator to induce phase separation and a liquid template to stabilize the superhydrophobic structure. The stable and durable superhydrophobic surface shows good biocompatibility and antibacterial properties.

Precise patterning of the SEBS surface by UV lithography to evaluate the platelet function through single platelet adhesion.

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.

Patterning surfaces for controlled platelet adhesion and detection of dysfunctional platelets.

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.

Measurement of adhesion work of electrospun polymer membrane by shaft-loaded blister test.

The work of adhesion at the interface of electrospun membrane and rigid substrate is measured by a shaft-loaded blister test (SLBT). Poly(vinylidene fluoride) (PVDF) were electrospun with an average fiber diameter of 333 ± 59 nm. Commercial cardboard with inorganic coating was used to provide a model substrate for adhesion tests. In SLBT, the elastic response PVDF was analyzed and its adhesion energy measured. The average value of the adhesion work is 206 ± 26 mJ/m(2). Elastic modulus of electrospun membrane obtained by SLBT is found to be 23.42 ± 2.69 MPa, which is consistent with the value obtained from standard tensile tests. The results show SLBT presented a viable methodology for evaluating the adhesion energy of electrospun polymer fabrics.

Mechanism of adhesion between polymer fibers at nanoscale contacts.

Adhesive force exists between polymer nano/microfibers. An elaborate experiment was performed to investigate the adhesion between polymer nano/microfibers using a nanoforce tensile tester. Electrospun polycaprolactone (PCL) fibers with diameters ranging from 0.4-2.2 µm were studied. The response of surface property of electrospun fiber to the environmental conditions was tracked by FTIR and atomic force microscopy (AFM) measurements. The effect of temperature on molecular orientation was examined by wide angle X-ray diffraction (WAXD). The adhesive force was found to increase with temperature and pull-off speed but insensitive to the change of relative humidity, and the abrupt increase of adhesion energy with temperature accompanied by a reduced molecular orientation in the amorphous part of fiber was observed. Results show that adhesion is mainly driven by van der Waals interactions between interdiffusion chain segments across the interface.