Thermoresponsive poly(N-isopropyl acrylamide)-grafted polycaprolactone films with surface immobilization of collagen

Thermoresponsive poly(N-isopropylacrylamide) (P(NIPAAm))-grafted polycaprolactone (PCL) films with a suitable amount of immobilized cell-adhesive collagen were prepared to improve cell adhesion and proliferation above the lower critical solution temperature (LCST, 32°C) of P(NIPAAm) without destroying cell detachment properties at lower temperatures. Covalently tethered P(NIPAAm) brushes on PCL film surfaces were first prepared via surface-initiated atom transfer radical polymerization (ATRP). The alkyl bromide end groups of the grafted P(NIPAAm) brushes were used in nucleophilic substitution reactions for the direct coupling of collagen to produce the collagen-immobilized thermoresponsive PCL surface. At 37°C, the cell attachments on the collagen-immobilized thermoresponsive PCL surface were enhanced substantially. The attached cells could be recovered simply by lowering culture temperature. The P(NIPAAm)-grafted PCL films with immobilized collagen are potentially useful as adhesion modifiers for advanced cell culture and tissue engineering applications.     

Preparation of molecularly imprinted polymer microspheres via atom transfer radical precipitation polymerization

The first combined use of atom transfer radical polymerization (ATRP) and precipitation polymerization in the molecular imprinting field is described. The utilized polymerization technique, namely atom transfer radical precipitation polymerization (ATRPP), provides MIP microspheres with obvious molecular imprinting effects towards the template, fast template binding kinetics and an appreciable selectivity over structurally related compounds. The living chain propagation mechanism in ATRPP results in MIP spherical particles with diameters (number‐average diameter D_n ≈ 3 μm) much larger than those prepared via traditional radical precipitation polymerization (TRPP). In addition, the MIP microspheres prepared via ATRPP have also proven to show significantly higher high‐affinity binding site densities on their surfaces than the MIP generated via TRPP, while the binding association constants K_a and apparent maximum numbers N_max of the high‐affinity sites as well as the specific template bindings are almost the same in the two cases. © 2009 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 47: 3257–3270, 2009     

One‐pot synthesis of surface‐functionalized molecularly imprinted polymer microspheres by iniferter‐induced “living” radical precipitation polymerization

This article describes for the first time the development of a new polymerization technique by introducing iniferter‐induced “living” radical polymerization mechanism into precipitation polymerization and its application in the molecular imprinting field. The resulting iniferter‐induced “living” radical precipitation polymerization (ILRPP) has proven to be an effective approach for generating not only narrow disperse poly(ethylene glycol dimethacrylate) microspheres but also molecularly imprinted polymer (MIP) microspheres with obvious molecular imprinting effects towards the template (a herbicide 2,4‐dichlorophenoxyacetic acid (2,4‐D)), rather fast template rebinding kinetics, and appreciable selectivity over structurally related compounds. The binding association constant K_a and apparent maximum number N_max for the high‐affinity sites of the 2,4‐D imprinted polymer were determined by Scatchard analysis and found to be 1.18 × 10⁴ M^?1 and 4.37 μmol/g, respectively. In addition, the general applicability of ILRPP in molecular imprinting was also confirmed by the successful preparation of MIP microspheres with another template (2‐chloromandelic acid). In particular, the living nature of ILRPP makes it highly useful for the facile one‐pot synthesis of functional polymer/MIP microspheres with surface‐bound iniferter groups, which allows their direct controlled surface modification via surface‐initiated iniferter polymerization and is thus of great potential in preparing advanced polymer/MIP materials. © 2010 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 48: 3217–3228, 2010     

Preparation and characterization of a thermoresponsive gigaporous medium for high-speed protein chromatography

A high-speed thermoresponsive medium was developed by grafting poly(N-isopropylacrylamide-co-butyl methacrylate) (P(NIPAM-co-BMA)) brushes onto gigaporous polystyrene (PS) microspheres via surface-initiated atom transfer radical polymerization (ATRP) technique, which has strong mechanical strength, good chemical stability and high mass transfer rate for biomacromolecules. The gigaporous structure, surface chemical composition, static protein adsorption, and thermoresponsive chromatographic properties of prepared medium (PS–P(NIPAM-co-BMA)) were characterized in detail. Results showed that the PS microspheres were successfully grafted with P(NIPAM-co-BMA) brushes and that the gigaporous structure was robustly maintained. After grafting, the nonspecific adsorption of proteins on PS microspheres was greatly reduced. A column packed with PS–P(NIPAM-co-BMA) exhibited low backpressure and significant thermo-responsibility. By simply changing the column temperature, it was able to separate three model proteins at the mobile phase velocity up to 2167 cm h⁻¹. In conclusion, the thermoresponsive polymer brushes grafted gigaporous PS microspheres prepared by ATRP are very promising in ‘green’ high-speed preparative protein chromatography.     

Efficient one‐pot synthesis of water‐compatible and photoresponsive molecularly imprinted polymer nanoparticles by facile RAFT precipitation polymerization

A facile, general, and highly efficient one‐pot approach to obtain azobenzene (azo)‐containing molecularly imprinted polymer (MIP) nanoparticles with photoresponsive template binding and release properties in aqueous media is described, which involves the combined use of hydrophilic macromolecular chain transfer agent‐mediated reversible addition‐fragmentation chain transfer precipitation polymerization and easily available water‐insoluble azo functional monomers. The resulting azo‐containing MIPs were characterized with dynamic laser scattering (DLS), SEM, FTIR, static contact angle and water dispersion studies, and equilibrium binding experiments. They have proven to be nanoparticles (their diameters being around 104–397 nm, as determined by DLS in methanol) with surface‐grafted hydrophilic polymer brushes and exhibit excellent pure water‐compatible template binding properties. Moreover, obvious photoregulated template binding behaviors were observed for such azo‐containing MIP nanoparticles, which led to their largely accelerated template release in the aqueous media under the UV light irradiation. Furthermore, the general applicability of the strategy was also demonstrated. © 2014 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2014 , 52, 1941–1952     

表面温敏聚合物刷的原子力显微镜和石英晶体微天平研究

通过表面引发的原子转移自由基聚合在硅片表面制备了聚（N-异丙基丙烯酰胺）（PNIPAAm）聚合物刷。用原子力显微镜（AFM）分别研究了PNIPAAm的接枝动力学、温度和溶剂性质对厚度的影响以及PNIPAAm链与原子力针尖间的粘附力。结果表明,PNIPAAm链在硅片表面的生长具有很好的可控性。常温下厚度为33nm的PNIPAAm膜在水溶液中的增加到82.4nm;而在甲醇/水（v/v,1:1）溶液中,PNIPAAm分子链处于坍塌收缩状态,厚度降低为45nm;在55℃下干燥所得厚度则仅为22nm。力-距离测量结果表明,在溶液中,PNIPAAm链与原子力针尖之间的粘附力远小于在干态下的粘附力。用石英晶体微天平（QCM-D）对PNIPAAm的可逆相转变进行了研究,结果表明PNIPAAm分子链随温度变化的构象转变是发生在30-34℃之间的连续过程。     

Fabrication of thermoresponsive polymer gradients for study of cell adhesion and detachment

A poly(N-isopropylacrylamide) (PNIPAAm) gradient covalently anchored on a silicon substrate with a linear variation of thickness was fabricated by continuous injection of the reaction mixture (NIPAAm, CuBr and its ligand, methanol, and water) into a glass chamber containing a silicon wafer, whose surface had been homogeneously immobilized with bromoisobutyryl bromide (BIBB). Because of the good control of the surface-initiated atom transfer radical polymerization (SI-ATRP) technique, the thickness of the PNIPAAm brushes was linearly proportional to the polymerization time. As a result, the gradient length and sharpness could be easily controlled by the experimental parameters such as the polymerization time and the injection rate. The as-prepared PNIPAAm gradients were characterized by ellipsometry, water contact angle, and atom force microscopy to detect their alteration of the thickness, surface wettability, and morphology, confirming the gradient structure. X-ray photoelectron spectroscopy confirmed the surface composition of the PNIPAAm. In vitro culture of HepG2 cells was implemented on the gradient surfaces, revealing that the cells could adhere at 37 degrees C and could be detached at 24 degrees C when the gradient thickness was in the range of 20-45 nm. The work thus develops a method to fabricate the stable gradient surface with better quality control, and clarifies in a facile manner the appropriate thickness of the PNIPAAm brushes in terms of cell adhesion and detachment.     

“Smart” molecularly imprinted monoliths for the selective capture and easy release of proteins

A new thermally switchable molecularly imprinted monolith for the selective capture and release of proteins has been designed. First, a generic poly(glycidyl methacrylate‐co‐ethylene dimethacrylate) monolith reacted with ethylenediamine followed by functionalization with 2‐bromoisobutyryl bromide to introduce the initiator for atom transfer radical polymerization. Subsequently, a protein‐imprinted poly(N‐isopropylacrylamide) layer was grafted onto the surface of the monolithic matrix by atom transfer radical polymerization. Scanning electron microscopy and energy‐dispersive X‐ray spectroscopy of the cross‐sections of imprinted monoliths confirmed the formation of dense poly(N‐isopropylacrylamide) brushes on the pore surface. The imprinted monolith exhibited high specificity and selectivity toward its template protein myoglobin over competing proteins and a remarkably large maximum adsorption capacity of 1641 mg/g. Moreover, this “smart” imprinted monolith featured thermally responsive characteristics that enabled selective capture and easy release of proteins triggered only by change in temperature with water as the mobile phase and avoided use of stronger organic solvents or change in ionic strength and pH.     

Synthesis of molecular imprinted organic-inorganic hybrid polymer binding caffeine

Random copolymers of poly{(methacrylamide)-co-(vinyl trimethoxysilane)} and poly{(methacrylic acid)-co-(vinyl trimethoxysilane)} were synthesized via a free radical polymerization reaction. Acid catalyzed sol-gel process of tetraethylorthosilicate (TEOS) with aforementioned polymers in the presence of methyl xanthine class of alkaloid like caffeine resulted in the formation of highly transparent monoliths. Solvent extraction of the template leaves behind the recognition sites intact with high selectivity towards the print molecule. The ionic and non-specific adsorptions, which are considered to be the main disadvantages of the molecularly imprinted polymers (MIP), are prevented to a considerable extent by the end capping of surface silanol groups. The template binding efficiencies of MIP were determined by HPLC analysis.     

Adsorption of carbaryl using molecularly imprinted microspheres prepared by precipitation polymerization

To obtain the desired specific adsorbents for carbaryl to enrichment, separation, and analysis of trace pesticide residues in environmental water, molecularly imprinted polymer (MIP) microspheres were prepared by precipitation polymerization using carbaryl, methacrylic acid (MAA), ethylene glycol dimethacrylate (EGDMA), azobisisobutyronitrile (AIBN), and acetonitrile as template, functional monomer, cross‐linker, initiator, and porogen, respectively. Molecular modeling software was used to compute rational interaction between the template molecule and function monomer. The adsorption properties of carbaryl in acetonitrile for imprinted microspheres were evaluated by equilibrium rebinding experiments. Scatchard plot analysis revealed that there was one class of binding sites populated in the imprinted polymer microspheres with dissociation constants of 3.3 × 10^?2 mol/l and an apparent maximum number of 1.95 µmol/g. The specificity of the imprinted microspheres was investigated by binding analysis using carbaryl and structurally related carbamate pesticides. The results indicated that the obtained imprinted microspheres showed a good selectivity for carbaryl. Copyright © 2007 John Wiley & Sons, Ltd.     

Preparation and application of molecularly imprinted polymer for isolation of chicoric acid from Chicorium intybus L. medicinal plant

Molecularly imprinted polymer (MIP) was synthesized and applied for the extraction of chicoric acid from Chicory herb (Chicorium intybus L.). A computational study was developed to find a suitable template to functional monomer molar ratio for MIP preparations. The molar ratio was chosen based on the comparison of the binding energy of the complexes between the template and functional monomers. Based on the computational results, eight different polymers were prepared using chicoric acid as the template. The MIPs were synthesized in a non-covalent approach via thermal free-radical polymerization, using two different polymerization methods, bulk and suspension. Batch rebinding experiments were performed to evaluate the binding properties of the imprinted polymers. The best results were obtained with a MIP prepared using bulk polymerization with 4-vinylpyridine (4-VP) as the functional monomer and ethylene glycol dimethacrylate (EGDMA) as the crosslinker with a molar ratio of 1:4:20. The best MIP showed selective binding ability toward chicoric acid in the presence of the template’s structural analogues, caffeic acid, caftaric acid and chlorogenic acid.     

Analytical followup of the gamma initiated synthesis of a molecularly imprinted polymer

A molecularly imprinted polymer (MIP) for the template phenytoin has been prepared by gamma initiated copolymerization of methacrylamide and ethylene glycol dimethacrylate. The progress of polymerization was studied by measuring the monomer conversions and the template binding properties of the resulting polymers, respectively. The consumption rate of the two monomers showed different course. There was no difference observed in the polymerization rates of the MIP and the control polymer (NIP). The template binding properties of the MIP and the NIP changed considerably with the progress of the polymerization process and became similar to those of the thermally initiated polymers after full conversion.     

Temperature-controlled flow switching in nanocapillary array membranes mediated by poly(N-isopropylacrylamide) polymer brushes grafted by atom transfer radical polymerization

We report actively controlled transport that is thermally switchable and size-selective in a nanocapillary array membrane (NCAM) prepared by grafting poly(N-isopropylacrylamide) (PNIPAAm) brushes onto the exterior surface of a Au-coated polycarbonate track-etched membrane. A smooth Au layer on the membrane surface, which is key to obtaining a uniform polymer film, was prepared by thermal evaporation of approximately 50 nm Au on both exterior surfaces. After evaporation, the inner diameter of the pore is reduced slightly, but the NCAM retains a narrow pore size distribution. PNIPPAm brushes with 10-30 nm (dry film) thickness were grafted onto the Au surface through surface-initiated atom transfer radical polymerization (ATRP) using a disulfide initiator, (BrC(CH3)2COO(CH2)11S)2. Molecular transport through the PNIPAAm polymer brush-modified NCAMs was investigated by real-time fluorescence measurements using fluorescein isothiocyanate (FITC)-labeled dextrans ranging from 4.4 to 282 kDa in membranes with variable initial pore diameters (80, 100, and 200 nm) and different PNIPAAm thicknesses. Manipulating the temperature of the NCAM through the PNIPAAm lower critical solution temperature (LCST) causes large, size-dependent changes in the transport rates. Over specific ranges of probe size, transport is completely blocked below the LCST but strongly allowed above the LCST. The combination of the highly uniform PNIPAAm brush and the monodisperse pore size distribution is critical in producing highly reproducible switching behavior. Furthermore, the reversible nature of the switching raises the possibility of using them as actively controlled filtration devices.     

Preparation of polysulfone‐g‐poly(N‐isopropylacrylamide) graft copolymers through atom transfer radical polymerization and formation of temperature‐responsive nanoparticles

Polysulfone‐g‐poly(N‐isopropylacrylamide) (PSf‐g‐PNIPAAm) graft copolymers were prepared from atom transfer radical polymerization of NIPAAm using chloromethylated PSf as a macro‐initiator. The chain lengths of PNIPAAm of the graft copolymers were controllable with polymerization reaction time. The chemical structures of the graft copolymers were characterized with FTIR, NMR, and elemental analysis and their amphiphilic characteristics were examined and discussed. The PSf‐g‐PNIPAAm graft copolymers and the nanoparticles made from the graft copolymers exhibited repeatable temperature‐responsive properties in heating–cooling cycles. © 2008 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 46: 4756–4765, 2008     

Preparation of molecularly imprinted polymers via atom transfer radical “bulk” polymerization

The first application of atom transfer radical “bulk” polymerization (ATRBP) in molecular imprinting is described, which provides molecularly imprinted polymers (MIPs) with obvious imprinting effects towards the template, very fast binding kinetics, and an appreciable selectivity over structurally related compounds. In comparison with the MIP prepared via the normally used traditional “bulk” free radical polymerization (BFRP), the MIPs obtained via ATRBP showed somewhat lower binding capacities and apparent maximum numbers N_max for high‐affinity sites as well as quite similar binding association constants K_a for high‐affinity sites and high‐affinity site densities, in contrast with the previous reports (e.g., nitroxide/iniferter‐mediated “bulk” polymerization provided MIPs with improved properties). This is tentatively ascribed to the occurrence of rather fast gelation process in ATRBP, which greatly restricted the mobility of the chemical species, leading to a heavily interrupted equilibrium between dormant species and active radicals and heterogeneous polymer networks. In addition, the general applicability of ATRBP was also confirmed by preparing MIPs for different templates. This work clearly demonstrates that applying controlled radical polymerizations (CRPs) in molecular imprinting not always benefits the binding properties of the resultant MIPs, which is of significant importance for the rational use of CRPs in generating MIPs with improved properties. © 2009 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 48: 532–541, 2010     

Efficient synthesis of narrowly dispersed molecularly imprinted polymer microspheres with multiple stimuli-responsive template binding properties in aqueous media

A facile, general, and highly efficient approach to obtain narrowly dispersed molecularly imprinted polymer microspheres with multiple stimuli-responsive template binding properties in aqueous media by successive RAFT polymerization is described.     

Gradient immobilization of a cell adhesion RGD peptide on thermal responsive surface for regulating cell adhesion and detachment

Using surface initiated atomic transfer radical polymerization (ATRP) and an injection method, a poly(N-isopropylacrylamide)-b-poly(acrylic acid)-g-RGD (PNIPAAm-b-PAA-g-RGD) gradient surface was prepared. First, a thermoresponsive surface with a constant thickness of PNIPAAm was fabricated, onto which the AA monomers were block copolymerized using the PNIPAAm macromolecules as initiators. During this process, a continuous injection method was employed to yield a molecular weight gradient of PAA on the underlying uniform PNIPAAm layer. RGD peptide was finally covalently immobilized onto the PAA gradient by carbodiimide chemistry. In vitro culture of HepG2 cells showed that immobilization of the RGD peptide could accelerate cell attachment, while the thermoresponsive layer beneath could effectively release the cells by simply lowering temperature. Thus, the PNIPAAm-b-PAA-g-RGD gradient surface, combining the thermal response with cell affinity properties, can well regulate the cell adhesion and detachment, which may thus be useful for investigation of cell-substrate interactions with a smaller number of samples.     

Covalent immobilization of antibody fragments on well-defined polymer brushes via site-directed method

Well-defined polymer brushes and block copolymer brushes consisting of 2-methacryloyloxyethyl phosphorylcholine (MPC) and glycidyl methacrylate (GMA) were prepared by surface-initiated atom transfer radical polymerization (ATRP). The polymer brushes were used for the immobilization of antibody fragments in a defined orientation. Pyridyl disulfide moieties were introduced to the polymer brushes via a reaction of epoxy groups in GMA units. Fab’ fragments were then immobilized onto these surfaces via a thiol-disulfide interchange reaction and the reactivity of antibodies with antigens was investigated. Antigen/antibody binding on the polymer brushes was more preferable than that on epoxysilane films as a control surface. Furthermore, the activity of the antibodies immobilized on the block copolymer brushes having biocompatible PMPC was greater than that on other surfaces that did not have PMPC in their structures.     

Synthesis and binding site characteristics of 2,4,6-trichlorophenol-imprinted polymers

2,4,6-Trichlorophenol (2,4,6-TCP)-imprinted micro- and submicrospheres prepared by precipitation polymerization were compared with templated materials obtained by conventional bulk polymerization. The influence of the type and amount of functional monomer, the type and amount of cross-linker, polymerization temperature, porogen, and the ratio of template molecule and functional monomer to cross-linker on the size of the obtained particles were investigated. UV-Vis spectrophotometer experiments revealed that the microsphere polymers provided higher affinity to the template in contrast to imprinted polymers prepared by bulk polymerization. The binding properties of the microspheres, including binding isotherms and affinity distribution, were studied via Freundlich isotherm affinity distribution (FIAD) analysis. The obtained results indicated that microspheres prepared by precipitation polymerization provided superior rebinding properties during equilibrium binding in contrast to bulk polymers and submicrosphere polymers. Moreover, release experiments showed that 80% of rebound 2,4,6-TCP was released from the imprinted microspheres within the first 2 h, while more intimately bound 2,4,6-TCP molecules were released in the following 40 h. The morphologies and porosities of the resulting imprinted materials were characterized by scanning electron microscopy (SEM) and Brunauer-Emmett-Teller (BET) analysis, respectively. The microsphere polymers exhibited a regular spherical shape with a high degree of monodispersity to the corresponding bulk polymers. Furthermore, the micro- and submicrospheres were characterized by narrow distribution of pores in contrast to a heterogeneity index of m = 0.6647 for the microsphere imprinted polymer.     

Thermoresponsive Double Network Hydrogels with Exceptional Compressive Mechanical Properties

The utility of thermoresponsive hydrogels, such as those based on poly(N‐isopropylacrylamide) (PNIPAAm), is severely limited by their deficient mechanical properties. In particular, the simultaneous achievement of high strength and stiffness remains unreported. In this work, a thermoresponsive hydrogel is prepared having the unique combination of ultrahigh compressive strength (≈23 MPa) and excellent compressive modulus (≈1.5 MPa). This is accomplished by employing a double network (DN) design comprised of a tightly crosslinked, highly negatively charged 1st network based on poly(2‐acrylamido‐2‐methylpropane sulfonic acid (PAMPS) and a loosely crosslinked, zwitterionic 2nd network based on a copolymer of thermoresponsive NIPAAm and zwitterionic 2‐(methacryloyloxy)ethyl]dimethyl‐(3‐sulfopropyl)ammonium hydroxide (MEDSAH). Comparison to other DN designs reveals that this PAMPS/P(NIPAAm‐co‐MEDSAH) DN hydrogel's remarkable properties stem from the intra‐ and internetwork ionic interactions of the two networks. Finally, this mechanically robust hydrogel retains the desirable thermosensitivity of PNIPAAm hydrogels, exhibiting a volume phase transition temperature of ≈35 °C.