Block Copolymer Brushes Research Articles

Inhibition of bacteria adhesion and biofilm formation using a precisely structured nitric oxide-releasing coating with repeatedly renewing antimicrobial and antifouling ability

Bacterial adhesion and colonization usually causes the formation of biofilm that cannot be easily eradicated by common antibiotics and disinfectants. Surface covalent immobilization of nitric oxide (NO)-releasing polymer is an effective surface modification strategy to combat the threat from bacteria. Although surface covalent strategy avoided the leaching of toxic NO donors, the existence of hydrophobic NO donors on topsurface could inevitably accelerate the microorganism adhesion and accumulation, and the NO-releasing period of time was still very limited. In this work, we prepared an antimicrobial and antibiofilm surface via tethering a precision-structured NO-releasing copolymer brush to an organic-inorganic hybrid hard coating (> 5H pencil hardness) on one end. The precision-structured diblock copolymer brush was composed of a surface antifouling block and a subsurface antimicrobial block of NO-releasing units. A typical effect of “kill two birds with one stone” was observed, the NO releasing subsurface segment within a polymeric matrix prolongs NO release while also preventing surface fouling caused by hydrophobic NO donors on top surface. The impacts of polymer architecture on bioactivity was detailed investigated, and the block copolymer brush exhibited the optimal inhibitory effects against bacteria colonization and biofilm formation. The developed bioactive diblock copolymer brush was grafted from a hard hybrid persistent coating that embeds considerable initiation sites for surface modification. Thanks to the presence of initiator throughout the hybrid network, the initiation layer could initiate polymerization repeatedly after the polymer brushes are worn off at high pressure. Thus, the coating exhibited repeatedly renewing antimicrobial and antifouling ability after multiple abrasion cycles. This work not only developed a novel strategy for regulating NO releasing via modulating the polymer architecture, but provided a feasible route for obtaining a robust initiator layer for synthesizing polymer brush for antimicrobial and antifouling applications.

Tailoring copolymer architectures and macromolecular interactions for enhanced nanotherapeutic delivery: A design-by-architecture approach

Amphiphilic copolymers with precise macromolecular topologies are crucial in nanomedicine for enhancing drug delivery by improving solubilization, stabilization, and therapeutic efficacy on target tissues. This study employs a design-by-architecture approach to identify factors influencing interactions between synthesized macromolecules and biological systems for optimal drug nanodelivery. Copolymers combining poly(ε-caprolactone) blocks and poly(polyethylene glycol methacrylate) or poly(glycerol methacrylate), were synthesized to investigate how architectural adjustments impact self-assembly, nanoparticle size, drug loading, and biomolecule interactions. Linear and brush block copolymers with varied block lengths were studied for their self-assembly behavior in aqueous environments. Both copolymer types formed smaller nanoparticles with extended hydrophilic blocks, with brush block copolymers showing superior performance due to their peculiar macromolecular architecture. Incorporating ‘clickable’ glycidyl units within the hydrophilic block minimally affected particle size. Controlled functionalization with thiol groups resulted in stable nanoparticles with enhanced size reduction and mucoadhesive properties, potentially improving targeted drug delivery and therapeutic effects.

Ionic nanoporous membranes from self-assembled liquid crystalline brush-like imidazolium triblock copolymers.

There is a need to generate mechanically and thermally robust ionic nanoporous membranes for separation and fuel cell applications. Herein, we report a general approach to the preparation of ionic nanoporous membranes through custom synthesis, self-assembly, and subsequent chemical manipulations of ionic brush block copolymers. We synthesized polynorbornene-based triblock copolymers containing imidazolium cations balanced by counter anions in the central block, side-chain liquid crystalline units, and sidechain polylactide end blocks. This unique platform comprises: (1) imidazolium/bis(trifluoromethanesulfonyl)imide (TFSI) as the middle block, which has an excellent ion-exchange ability, (2) cyanobiphenyl liquid crystalline end block, a sterically hindered hydrophobic segment, which is chemically stable and immune to hydroxide attack, (3) polylactide brush-like units on the other end block that is easily etched under mild alkaline conditions and (4) a polynorbornene backbone, a lightly crosslinked system that offers mechanical robustness. These membranes retain their morphology before and after backbone crosslinking as well as etching of polylactide sidechains. The ion exchange performance and dimensional stability of these membranes were investigated by water uptake capability and swelling ratio. Moreover, the length of the carbon spacer in the imidazolium/TFSI central block moiety endowed the membrane with improved ionic conductivity. The ionic nanoporous materials are unusual due to their singular thermal, mechanical, alkaline stability and ion transport properties. Applications of these materials include electrochemical actuators, solid-state ionic nanochannel biosensors, and ion-conducting membranes.

Surface-Grafted Biocompatible Polymer Conductors for Stable and Compliant Electrodes for Brain Interfaces.

Durable and conductive interfaces that enable chronic and high-resolution recording of neural activity are essential for understanding and treating neurodegenerative disorders. These chronic implants require long-term stability and small contact areas. Consequently, they are often coated with a blend of conductive polymers and are crosslinked to enhance durability despite the potentially deleterious effect of crosslinking on the mechanical and electrical properties. Here the grafting of the poly(3,4 ethylenedioxythiophene) scaffold, poly(styrenesulfonate)-b-poly(poly(ethylene glycol) methyl ether methacrylate block copolymer brush to gold, in a controlled and tunable manner, by surface-initiated atom-transfer radical polymerization (SI-ATRP) is described. This "block-brush" provides high volumetric capacitance (120 F cm─3), strong adhesion to the metal (4h ultrasonication), improved surface hydrophilicity, and stability against 10000 charge-discharge voltage sweeps on a multiarray neural electrode. In addition, the block-brush film showed 33% improved stability against current pulsing. This approach can open numerous avenues for exploring specialized polymer brushes for bioelectronics research and application.

Chiral polymer coatings on substrates via surface-initiated RAFT polymerization under ambient conditions

Surface-initiated reversible addition-fragmentation chain transfer polymerization using oxygen (O2-SI-RAFT) was performed to endow the substrates with chiral properties. Chiral monomers were synthesized using methacrylic anhydride and l/d-amino acids (phenylalanine methyl ester, alanine methyl ester, valine methyl ester, and leucine methyl ester), and chiral polymers were grown on titanium and glass substrates. The polymer brushes grafted from the substrates were confirmed by ellipsometry analysis, water contact angles, X-ray photoelectron spectroscopy, and atomic force microscopy. Block copolymer brushes were also polymerized through chain extension. Using a glass substrate, chirality of the polymer brushes grown on the surface could be confirmed via circular dichroism spectroscopy.

A thermoresponsive cationic block copolymer brush-grafted silica bead interface for temperature-modulated separation of adipose-derived stem cells

Adipose-derived mesenchymal stem cells (ADSCs) have beneficial effects in cell transplantation therapy; these cells are collected from adipose tissue using low-invasive methods. However, to prepare ADSCs for cell therapy, a cell separation method that neither involves modification of the cell surface nor causes loss of cell activity is needed. Here, we aimed to develop ADSC separation columns using thermoresponsive cationic block copolymer brush-grafted beads as packing materials. The block copolymer brush was formed by a bottom cationic segment, poly(N,N-dimethylaminopropylacrylamide) (PDMAPAAm), and an upper thermoresponsive segment, poly(N-isopropylacrylamide) (PNIPAAm), and was grafted in two atom transfer radical polymerization reactions. The copolymer brush-grafted silica beads were packed into a column. An ADSC suspension was introduced into the columns at 37 °C and adsorbed on the copolymer brush-modified beads through electrostatic and hydrophobic interactions with the PDMAPAAm and PNIPAAm segments, respectively. The adsorbed ADSCs eluted from the column by lowering the temperature to 4 °C. In contrast, most Jurkat and vascular endothelial cells eluted at 37 °C, because of the relatively weaker electrostatic interactions with the block copolymer brush compared to ADSCs. Using the prepared column, a mixture of ADSCs and Jurkat cells was separated by changing the column temperature. The recovered ADSCs exhibited cell activity. The developed cell separation column may be useful for isolating ADSCs without cell surface modification, while maintaining cell activity.

Functional Poly(ε-caprolactone)/Poly(ethylene glycol) Copolymers with Complex Topologies for Doxorubicin Delivery to a Proteinase-Rich Tumor Environment

Doxorubicin (DOX)-loaded polymer nanoparticles based on poly(ethylene glycol)-poly(ε-caprolactone) copolymers with a complex macromolecular topology are proposed to tackle the matrix metalloproteinase (MMP)-rich tumor environment. Linear, 4-arm comb-like copolymers and 4-arm brush block copolymers were synthesized through a combination of ring opening polymerization and atom transfer radical polymerization, in order to control the molar mass distribution, the arm/brush architecture, as well as the final size and DOX loading of self-assembled nanoparticles obtained by nanoprecipitation. The optimized nanocarriers were conjugated with penetrating low molecular weight protamine peptides coupled to a polyanionic inhibitory domain cleavable by matrix metalloproteinase-2 (MMP2). DOX-loaded, MMP2-activable nanocarriers were evaluated in the context of glioblastoma (GBM), a brain tumor characterized by remarkable and relevant MMP2 expression. Uptake and cytotoxicity in patient-derived GBM cells correlated with the level of MMP2 enzymatic activity in a dose- and time-dependent manner. No effects were observed in nontumoral endothelial cells that do not express MMP2. Results demonstrated that, by tuning polymer topology and peptide sequence, nanoparticle self-assembly, DOX encapsulation, and delivery can be optimized for the development of an advanced treatment for MMP2-overexpressing tumors.

A Facile Fabrication of CdSe/ZnS QDs-Block Copolymer Brushes-Modified Graphene Oxide Nanohybrid with Temperature-Responsive Behavior.

In this paper, we described a straightforward one-step chemical method for the synthesis of semiconductor quantum dots(QDs)—block copolymer brushes functionalized graphene oxide(GO) fluorescence nanohybrids. The azobenzene-terminated block copolymer poly(N-isopropylacrylamid)-b-poly(styrene-co-5-(2-methacryoylethyloxymethyl)-8-quinolinol)(PNIPAM-b-P(St-co-MQ)) was modified on the surface of GO sheets via host–guest interactions between β-cyclodextrin-modified GO and azobenzene moieties, and simultaneously CdSe/ZnS QDs were integrated on the block copolymer brushes through the coordination between 8-hydroxyquinoline units in the polymer brushes and CdSe/ZnS QDs. The resulting fluorescence nanohybrid exhibited dual photoluminescence at 620 nm and 526 nm, respectively, upon excitation at 380 nm and LCST-type thermo-responsive behavior which originated from the change in the PNIPAM conformation in the block copolymer brushes of GO sheets.

Multifunctional Photonic Janus Particles.

Photonic Janus particles with a sphere fused to a cone are created from the phase separation of dendronized brush block copolymers (den-BBCP) and poly(4-vinylpyridine)-r-polystyrene (P4VP-r-PS) during the solvent evaporation of oil-in-water emulsions. Rapid self-assembly of den-BBCP generates well-ordered lamellar structures stacking along the long axis of the particles, producing structural colors that are dependent on the incident light angle. The colors are tunable over the visible spectrum by varying the molecular weight of den-BBCP. The P4VP-r-PS phase can undergo further surface modifications to produce multifunctional photonic Janus particles. Specifically, real-time magnetic control of the reflected color is achieved by coating the P4VP-r-PS phase with citric acid-capped Fe3O4 nanoparticles. Charged biomolecules (i.e., antibodies) are electrostatically immobilized to the Fe3O4 coating for potential applications in biosensing. As a demonstration, a new photonic sensor for the foodborne pathogen Salmonella is developed with antibody-modified photonic Janus particles, where the angle-dependent structural color plays a key role in the sensing mechanism.

Synthesis and Self-Assembly of Poly(vinylpyridine)-Containing Brush Block Copolymers: Combined Synthesis of Grafting-Through and Grafting-to Approaches

Synthesis and Self-Assembly of Poly(vinylpyridine)-Containing Brush Block Copolymers: Combined Synthesis of Grafting-Through and Grafting-to Approaches

Enhanced vapor sorption in block and random copolymer brushes.

Polymer brushes in gaseous environments absorb and adsorb vapors of favorable solvents, which makes them potentially relevant for sensing applications and separation technologies. Though significant amounts of vapor are sorbed in homopolymer brushes at high vapor pressures, at low vapor pressures sorption remains limited. In this work, we vary the structure of two-component polymer brushes and investigate the enhancement in vapor sorption at different relative vapor pressures compared to homopolymer brushes. We perform molecular dynamics simulations on two-component block and random copolymer brushes and investigate the influence of monomer miscibility and formation of high-energy interfaces between immiscible monomers on vapor sorption. Additionally, we present absorption isotherms of pure homopolymer, mixed binary brush and 2-block, 4-block, and random copolymer brushes. Based on these isotherms, we finally show that random copolymer brushes absorb more vapor than any other architecture investigated thus far. Random brushes display enhanced sorption at both high and low vapor pressures, with the largest enhancement in sorption at low vapor pressures.

Thermoresponsive block copolymer brush for temperature-modulated hepatocyte separation.

Hepatic tissue engineering may be an effective approach for the treatment of liver disease; however, its practical application requires hepatic cell separation technologies that do not involve cell surface modification and maintain cell activity. In this study, we developed hepatocyte cell separation materials using a thermoresponsive polymer and a polymer with high affinity to hepatocytes. A block copolymer of poly(N-p-vinylbenzyl-O-β-D-galactopyranosyl-(1→4)-D-gluconamide) (PVLA) and poly(N-isopropylacrylamide) (PNIPAAm) [PVLA-b-PNIPAAm] was prepared through two steps of atom transfer radical polymerization. On the prepared PVLA-b-PNIPAAm brush, HepG2 cells (model hepatocytes) adhered at 37 °C and detached at 20 °C, attributed to the temperature-modulated affinity between PVLA and HepG2. Cells from the immortalized human hepatic stellate cell line (TWNT-1) did not adhere to the copolymer brush, and RAW264.7 cells (mouse macrophage; model Kupffer cells) adhered to the copolymer brush, regardless of temperature. Using the difference in cell adhesion properties on the copolymer brush, temperature-modulated cell separation was successfully demonstrated. A mixture of HepG2, RAW264.7, and TWNT-1 cells was seeded on the copolymer brush at 37 °C for adherence. By reducing the temperature to 20 °C, adhered HepG2 cells were selectively recovered with a purity of approximately 85% and normal activity. In addition, induced pluripotent stem (iPS) cell-derived hepatocytes adhered on the PVLA-b-PNIPAAm brush at 37 °C and detached from the copolymer brush at 20 °C, whereas the undifferentiated iPS cells did not adhere, indicating that the prepared PVLA-b-PNIPAAm brush could be utilized to separate hepatocyte differentiated and undifferentiated cells. These results indicated that the newly developed PVLA-b-PNIPAAm brush can separate hepatic cells from contaminant cells by temperature modulation, without affecting cell activity or modifying the cell surface. Thus, the copolymer brush is expected to be a useful separation tool for cell therapy and tissue engineering using hepatocytes.

Near-Infrared Fluorescent Micelles from Poly(norbornene) Brush Triblock Copolymers for Nanotheranostics.

This contribution describes the design and synthesis of multifunctional micelles based on amphiphilic brush block copolymers (BBCPs) for imaging and selective drug delivery of natural anticancer compounds. Well-defined BBCPs were synthesized via one-pot multi-step sequential grafting-through ring-opening metathesis polymerization (ROMP) of norbornene-based macroinitiators. The norbornenes employed contain a poly(ethylene glycol) methyl ether chain, an alkyl bromide chain, and/or a near-infrared (NIR) fluorescent cyanine dye. After block copolymerization, post-polymerization transformations using bromide-azide substitution, followed by the strain-promoted azide-alkyne cycloaddition (SPAAC) allowed for the functionalization of the BBCPs with the piplartine (PPT) moiety, a natural product with well-documented cytotoxicity against cancer cell lines, via an ester linker between the drug and the polymer side chain. The amphiphilic BBCPs self-assembled in aqueous media into nano-sized spherical micelles with neutral surface charges, as confirmed by dynamic light scattering analysis and transmission electron microscopy. During self-assembly, paclitaxel (PTX) could be effectively encapsulated into the hydrophobic core to form stable PTX-loaded micelles with high loading capacities and encapsulation efficiencies. The NIR fluorescent dye-containing micelles exhibited remarkable photophysical properties, excellent colloidal stability under physiological conditions, and a pH-induced disassembly under slightly acidic conditions, allowing for the release of the drug in a controlled manner. The in vitro studies demonstrated that the micelles without the drug (blank micelles) are biocompatible at concentrations of up to 1 mg mL-1 and present a high cellular internalization capacity toward MCF-7 cancer cells. The drug-functionalized micelles showed in vitro cytotoxicity comparable to free PPT and PTX against MCF-7 and PC3 cancer cells, confirming efficient drug release into the tumor environment upon cellular internalization. Furthermore, the drug-functionalized micelles exhibited higher selectivity than the pristine drugs and preferential cellular uptake in human cancer cell lines (MCF-7 and PC3) when compared to the normal breast cell line (MCF10A). This study provides an efficient strategy for the development of versatile polymeric nanosystems for drug delivery and image-guided diagnostics. Notably, the easy functionalization of BBCP side chains via SPAAC opens up the possibility for the preparation of a library of multifunctional systems containing other drugs or functionalities, such as target groups for recognition.

A general and robust strategy for in-situ templated synthesis of patterned inorganic nanoparticle assemblies

Precisely controlling the spatial organization of inorganic nanoparticles is essential for many applications, such as sensing, catalytic, and optical/electronic systems. Here, we describe a general and robust strategy to fabricate arbitrary quasi-two-dimensional (quasi-2D) nanoparticle assembly structures with tunable size and thickness on a substrate by using block copolymer brushes as the templates. To achieve this, patterned amphiphilic block copolymer brushes were first grown via light-mediated surface-initiated atom transfer radical polymerization. The hydrophilic polymer blocks could interact and enrich the inorganic precursors, and thus accelerate the in-situ growth of nanoparticles within the patterned hydrophilic polymer domains. In our method, the block copolymer template could strictly limit the vertical growth of quasi-2D nanoparticle assemblies to achieve thickness control in the nanometer scale. By changing the design of the photomask and experimental conditions for the light-mediated polymerization, quasi-2D inorganic nanoparticle assemblies with controllable shape and dimension can be feasibly obtained. Moreover, this method is also generally applicable to prepare various patterned inorganic nanoparticle assemblies with different compositions.

Synthesis and micellar property of amphiphilic brush-arm star copolymers via living ROMP

Norbornene macromonomers (MMs) with polystyrene (PS) or poly(tert-butyl acrylate) (PtBA) side chains were synthesized by a combination of atom-transfer radical polymerization (ATRP) and click reactions. A series of amphiphilic linear and star-shaped brush block copolymers were obtained through sequential ring opening metathesis polymerization (ROMP) of MMs by the use of the highly active olefin metathesis Blechert's catalyst and tri-functionalized Blechert's catalyst respectively, followed by hydrolysis reaction. These well-defined diblock molecular brushes can undergo further self-assembly to form spherical micelles in solution, which have been characterized using DLS, AF4-MALLS, AFM, TEM and fluorescence spectra. The dynamic diameter increases as the hydrophilic backbone content increases for the vast majority of the linear and star copolymers. Brush-arm star diblock amphiphilic copolymers have smaller hydrodynamic diameter and aggregation number than the corresponding linear analogues. Moreover, the critical micelle concentration (CMC) values of star diblock copolymers show less sensitive to the variety of composition.

Graphitic Mesoporous Carbon Thin Film Prepared By Rapid Thermal Annealing for Micro-Supercapacitors

The miniaturization of electronic devices led to the advent of on-chip power sources with high energy and power density requirement. The development of such miniaturized energy devices further requires the preparation and/or processing of thin films (down to a few nanometers) of active material. Mesoporous carbon possessing both high porosity and conductivity are desired for such applications. However, obtaining thin films of mesoporous carbon on various substrates using commercially viable approaches requires further improvement and investigation. We have adapted rapid thermal annealing (RTA) as a novel approach to obtain mesoporous carbon. RTA enables rapid heating to temperatures up to 1200oC in just a few seconds and therefore provides a route for complete carbonization within a few minutes. Carbon precursor films comprised of brush block copolymers (BBCP) as sacrificial templates and phenol formaldehyde (PF) resin as a carbon source were prepared. In particular, the self-assembly of polydimethylsiloxane-b-polyethylene oxide (PDMS–b-PEO) BBCP and PF resin led to the formation of microphase-segregated film on various substrates including Si wafer, fused silica, and stainless steel. The PDMS domain in the BBCP fostered strong adhesion to the substrate even after annealing. RTA enabled complete carbonization of the precursor film within only 5 minutes, whereas conventional methods require approximately 500 minutes. Rapid processing at 1000oC resulted in the formation of mesoporous carbon with higher degree of graphitization, which is difficult to achieve with conventional carbonization method. The device prepared via RTA has a capacitance value of 7.0 mF/cm2 at 100mV/s, which is one of the highest values obtained for nanometer-thick carbon-based electrodes. These electrodes maintained a quasi-rectangular shape during the potential vs current scan, even at a scan rate of 20 V/s, which suggests that these materials are promising candidates for applications requiring very high-power density.The mesoporous carbon structure having high porosity and high conductivity can be further utilized as supports for the deposition of pseudo-capacitance materials to further boost the energy density of the device. Figure 1

Polymer Coupling via Hetero-Disulfide Exchange and Its Applications to Rewritable Polymer Brushes.

An iodide-terminated polymer (Polymer-I) is converted to a thiol-terminated polymer (Polymer-SH) using HSCH2CH2SH in a remarkably short time (10 min). Polymer-SH is further converted to a pyridyl disulfide-terminated polymer (Polymer-SS-Py). The hetero-coupling of Polymer-SH and Polymer-SS-Py is successfully achieved to quantitatively generate a polymer disulfide (Polymer-SS-Polymer). Exploiting this efficient hetero-coupling technique, Polymer-SH is attached (grafted) on a Py-SS-immobilized surface to generate a polymer brush via a disulfide (-SS-) linkage (writing process). The -SS- linkage is cleaved by the treatment with dithiothreitol (DTT) to detach the polymer from the surface (erasing process). Subsequently, another Polymer-SH is attached on the surface to generate another polymer brush (rewriting process). Thus, a writable, erasable, and rewritable polymer brush surface is achieved. Hydrophilic, hydrophobic, and super-hydrophobic polymers (Polymer-SH) are attached on the surface, tailoring the surface wettability in the writing-erasing-rewriting cycles. Polymer-SH is also attached on a chain-end Py-SS-functionalized polymer brush surface, generating a rewritable block copolymer brush surface. A patterned block copolymer brush surface is also obtained using photo-irradiation and a photo-mask in the erasing process. The metal-free synthetic procedure, accessibility to patterned brushes, and switchable surface properties via the writing-erasing-rewriting process are attractive features of the present approach.

Precise positioning of enzymes within hierarchical polymer nanostructures for switchable bioelectrocatalysis

The ability to reversibly switch bioelectrocatalytic sensors is attractive for the design of biomonitoring platforms displaying a complex environmental response, or for the protection of biosensors. However, the retention of reversible biocatalytic properties upon multiple environmental cycles, with broad detection range, low signal-to-noise and limit of detection remains challenging. In this report, we demonstrate the precise positioning of the enzyme glucose oxidase within block-copolymer brush nanostructures, via the re-initiation of N-isopropylacrylamide (NIPAM) polymerisation from enzyme-decorated poly(dimethylaminoethyl methacrylate) (PDMAEMA) blocks. We find that the precise design of polymer brush grafting density, thickness and crosslinking of the PNIPAM block enables the stable positioning of biocatalytic sites close to electrode surfaces. The control of the polymer brush nanostructure, its conformation and the distribution of biocatalytic sites is characterised via a combination of in situ ellipsometry, X-ray photoelectron spectroscopy, grazing angle FTIR and surface plasmon resonance. In turn, cyclic voltammetry and electrochemical impedance spectroscopy demonstrate that such control of the polymeric nanostructures confers a unique combination of low limit of detection (23.9μM), a broad dynamic range of glucose sensing (0.05-12.8mM) and true "OFF" state upon pH or thermal stimulation, whilst retaining excellent performance over repeated switching cycles of the sensor. Therefore, hierarchical biocatalytic polymer brushes display unique properties for the design of responsive biosensors and complex multi-functional gating platforms.

Thermally-modulated cell separation columns using a thermoresponsive block copolymer brush as a packing material for the purification of mesenchymal stem cells.

Cell therapy using mesenchymal stem cells (MSCs) is used as effective regenerative treatment. Cell therapy requires effective cell separation without cell modification and cellular activity reduction. In this study, we developed a temperature-modulated mesenchymal stem cell separation column. A temperature-responsive cationic block copolymer, poly(N,N-dimethylaminopropylacrylamide)-b-poly(N-isopropylacrylamide)(PDMAPAAm-b-PNIPAAm) brush with various cationic copolymer compositions, was grafted onto silica beads via two-step atom transfer radical polymerization. Using the packed beads, the elution behavior of the MSCs was observed. At 37 °C, the MSCs were adsorbed onto the column via both hydrophobic and electrostatic interactions with the PNIPAAm and PDMAPAAm segments of the copolymer brush, respectively. By reducing the temperature to 4 °C, the adsorbed MSCs were eluted from the column by reducing the hydrophobic and electrostatic interactions attributed to the hydration and extension of the PNIPAAm segment of the block copolymer brush. From the temperature-modulated adsorption and elution behavior of MSCs, a suitable DMAPAAm composition of the block copolymer brush was determined. Using the column, a mixture of MSC and BM-CD34+ cells was separated by simply changing the column temperature. The column was used to purify the MSCs, with purities of 78.2%, via a temperature change from 37 °C to 4 °C. Additionally, the cellular activity of the MSCs was retained throughout the column separation step. Overall, the obtained results show that the developed column is useful for MSC separation without cell modification and cellular activity reduction.

Selective capture and non-invasive release of cells using a thermoresponsive polymer brush with affinity peptides.

Tissue engineering and cell transplantation therapy have become promising therapies for intractable diseases. These approaches require cell separation technology without cell modification. Accordingly, in this study, we developed a novel cell separation method using a thermoresponsive block copolymer brush with an affinity peptide. A block copolymer brush with bottom poly(2-hydroxyethyl methacrylate [HEMA]-co-propargyl acrylate) and top poly(N-isopropylacrylamide-co-HEMA) segments was prepared through two steps of atom transfer radical polymerization. Then, cell affinity peptides were conjugated to the bottom segment of the copolymer brush through a click reaction. Using cRGD as a cell-affinity peptide, enhancement of cell adhesion with rapid adhesion on the copolymer brush was observed at 37 °C, whereas the copolymer brush without cRGD did not exhibit cell adhesion. Temperature-modulated cell adhesion and detachment were performed with a relatively long upper segment because the affinity between peptides and cells was modulated by the swelling and shrinking of the upper thermoresponsive segment. Selective endothelial cell adhesion was performed at 37 °C using GGGREDV as an affinity peptide. Smooth muscle cells and fibroblasts did not adhere to the copolymer brush. Adhered human umbilical vein endothelial cells (HUVECs) were successfully recovered by reducing the temperature to 20 °C. Based on the properties of the copolymer brush, HUVECs could be purified using a mixture of cells simply by changing the temperature. These results demonstrated that the prepared copolymer brush with cell affinity peptides could be a useful cell separation tool because the cells could be separated with specificity and without cell modification using a simple procedure.