Antifouling Polymer Brushes Research Articles

In Situ Growth of Columnar PEG on PEDOT and Its Antifouling Properties.

The antifouling properties of conductive polymers have received extensive attention for biosensor and bioelectronic applications. Polyethylene glycol (PEG) is a well-known antifouling material, but the controlled regulation of the surface topography of PEG without a template remains a challenge. Here, we show a columnar structure antifouling conductive polymer brush with enhanced antifouling properties and considerable conductivity. The method involves synthesizing the 3,4-ethylenedioxythiophene monomer modified with azide (EDOT-N3), the electropolymerization of PEDOT-N3, and the in situ growth of PEG polymer brushes on PEDOT through double-click reactions. The resultant columnar structure polymer brush exhibits high electrical conductivity (3.5 Ω·cm2), ultrahigh antifouling property, electrochemical stability (capacitance retention was 93.8% after 2000 cycles of CV scans in serum), and biocompatibility.

Study of Hydration Repulsion of Zwitterionic Polymer Brushes Resistant to Protein Adhesion through Molecular Simulations.

The fabrication of antifouling zwitterionic polymer brushes represents a leading approach to mitigate nonspecific adhesion on the surfaces of medical devices. This investigation seeks to elucidate the correlation between the material composition and structural attributes of these polymer brushes in preventing protein adhesion. To achieve this goal, we modeled three different zwitterionic brushes, namely, carboxybetaine methacrylate (CBMA), sulfobetaine methacrylate (SBMA), and (2-(methacryloyloxy)ethyl)-phosphorylcholine (MPC). The simulations revealed that elevating the grafting density enhances the structural stability, hydration strength, and resistance to protein adhesion exhibited by the polymer brushes. PCBMA manifests a more robust hydration layer, while PMPC demonstrates the slightest interaction with proteins. In a comprehensive evaluation, PSBMA polymer brushes emerged as the best choice with superior stability, enhanced protein repulsion, and minimally induced protein deformation, resulting in effective resistance to nonspecific adhesion. The high-density SBMA polymer brushes significantly reduce the level of protein adhesion in AFM testing. In addition, we have pioneered the quantitative characterization of hydration repulsion in polymer brushes by analyzing the hydration repulsion characteristics at different materials and graft densities. In summary, our study provides a nuanced understanding of the material and structural determinants influencing the capacity of zwitterionic polymer brushes to thwart protein adhesion. Additionally, it presents a quantitative elucidation of hydration repulsion, contributing to the advancement and application of antifouling polymer brushes.

Sandwich Immuno-RCA Assay with Single Molecule Counting Readout: The Importance of Biointerface Design.

The analysis of low-abundance protein molecules in human serum is reported based on counting of the individual affinity-captured analyte on a solid sensor surface, yielding a readout format similar to digital assays. In this approach, a sandwich immunoassay with rolling circle amplification (RCA) is used for single molecule detection (SMD) through associating the target analyte with spatially distinct bright spots observed by fluorescence microscopy. The unspecific interaction of the target analyte and other immunoassay constituents with the sensor surface is of particular interest in this work, as it ultimately limits the performance of this assay. It is minimized by the design of the respective biointerface and thiol self-assembled monolayer with oligoethylene (OEG) head groups, and a poly[oligo(ethylene glycol) methacrylate] (pHOEGMA) antifouling polymer brush was used for the immobilization of the capture antibody (cAb) on the sensor surface. The assay relying on fluorescent postlabeling of long single-stranded DNA that are grafted from the detection antibody (dAb) by RCA was established with the help of combined surface plasmon resonance and surface plasmon-enhanced fluorescence monitoring of reaction kinetics. These techniques were employed for in situ measurements of conjugating of cAb to the sensor surface, tagging of short single-stranded DNA to dAb, affinity capture of the target analyte from the analyzed liquid sample, and the fluorescence readout of the RCA product. Through mitigation of adsorption of nontarget molecules on the sensor surface by tailoring of the antifouling biointerface, optimizing conjugation chemistry, and by implementing weak Coulombic repelling between dAb and the sensor surface, the limit of detection (LOD) of the assay was substantially improved. For the chosen interleukin-6 biomarker, SMD assay with LOD at a concentration of 4.3 fM was achieved for model (spiked) samples, and validation of the ability of detection of standard human serum samples is demonstrated.

Perspectives on Theoretical Models and Molecular Simulations of Polymer Brushes.

Polymer brushes have witnessed extensive utilization and progress, driven by their distinct attributes in surface modification, tethered group functionality, and tailored interactions at the nanoscale, enabling them for various scientific and industrial applications of coatings, sensors, switchable/responsive materials, nanolithography, and lab-on-a-chips. Despite the wealth of experimental investigations into polymer brushes, this review primarily focuses on computational studies of antifouling polymer brushes with a strong emphasis on achieving a molecular-level understanding and structurally designing antifouling polymer brushes. Computational exploration covers three realms of thermotical models, molecular simulations, and machine-learning approaches to elucidate the intricate relationship between composition, structure, and properties concerning polymer brushes in the context of nanotribology, surface hydration, and packing conformation. Upon acknowledging the challenges currently faced, we extend our perspectives toward future research directions by delineating potential avenues and unexplored territories. Our overarching objective is to advance our foundational comprehension and practical utilization of polymer brushes for antifouling applications, leveraging the synergy between computational methods and materials design to drive innovation in this crucial field.

Durable, Ultrathin, and Antifouling Polymer Brush Coating for Efficient Condensation Heat Transfer.

Heat exchangers are made of metals because of their high heat conductivity and mechanical stability. Metal surfaces are inherently hydrophilic, leading to inefficient filmwise condensation. It is still a challenge to coat these metal surfaces with a durable, robust, and thin hydrophobic layer, which is required for efficient dropwise condensation. Here, we report the nonstructured and ultrathin (∼6 nm) polydimethylsiloxane (PDMS) brushes on copper that sustain high-performing dropwise condensation in high supersaturation. Due to the flexible hydrophobic siloxane polymer chains, the coating has low resistance to drop sliding and excellent chemical stability. The PDMS brushes can sustain dropwise condensation for up to ∼8 h during exposure to 111 °C saturated steam flowing at 3 m·s-1, with a 5-7 times higher heat transfer coefficient compared to filmwise condensation. The surface is self-cleaning and can reduce the level of bacterial attachment by 99%. This low-cost, facile, fluorine-free, and scalable method is suitable for a great variety of heat transfer applications.

Formation of Antifouling Brushes on Various Substrates Using a Melanin-Inspired Initiator Film.

In this study, we developed a substrate-independent initiator film that can undergo surface-initiated polymerization to form an antifouling brush. Inspired by the melanogenesis found in nature, we synthesized a tyrosine-conjugated bromide initiator (Tyr-Br) that contains phenolic amine groups as the dormant coating precursor and α-bromoisobutyryl groups as the initiator. The resultant Tyr-Br was stable under ambient air conditions and underwent melanin-like oxidation only in the presence of tyrosinase to form an initiator film on various substrates. Subsequently, an antifouling polymer brush was formed using air-tolerant activators regenerated by electron transfer for atom transfer radical polymerization (ARGET ATRP) of zwitterionic carboxybetaine. The entire surface coating procedure, including the initiator layer formation and ARGET ATRP, occurred under aqueous conditions and did not require organic solvents or chemical oxidants. Therefore, antifouling polymer brushes can be feasibly formed not only on experimentally preferred substrates (e.g., Au, SiO2, and TiO2) but also on polymeric substrates such as poly(ethylene terephthalate) (PET), cyclic olefin copolymer (COC), and nylon.

Grafting of Porous Conductive Fiber Mats with an Antifouling Polymer Brush by Means of Filtration-Based Surface Initiated ATRP.

This work addresses the challenge of surface modification of porous, electrospun fiber mats containing an insoluble conducting polymer coating. Herein, a novel methodology of grafting a polymer brush onto conducting polymer fiber mats is developed that employs filtering of the polymerization solution through the fiber mat. An electrospun sulfonated polystyrene-poly(ethylene-ran-butylene)-polystyrene (sSEBS) fiber mat is first coated with a layer of conducting copolymer bearing an Atom Transfer Radical Polymerization (ATRP) initiating functionality (PEDOT-Br). The surface-initiated ATRP from the fibers' surface is then carried out to graft a hydrophilic polymer brush (poly(ethylene glycol) methyl ether methacrylate) by means of filtering the polymerization solution through the fiber mat. Scanning electron microscopy (SEM) images reveal a progressive change in the morphology of the fiber mat surface with the increasing volume of the filtrated polymerization solution, while energy dispersive X-ray spectrosdcopy (EDX) spectra show a change in the atomic oxygen to sulfur (O/S) ratio, therefore confirming the successful grafting from the fibers' surface. The conductive fiber mat grafted with hydrophilic brushes shows a 20% reduction in the non-specific adsorption of bovine serum albumin (BSA) compared to a pristine fiber mat. This study is a proof-of-concept for this novel, filtration-based, surface-initiated polymerization methodology.

The Relation Between Protein Adsorption and Hemocompatibility of Antifouling Polymer Brushes.

Whenever an artificial surface comes into contact with blood, proteins are rapidly adsorbed onto its surface. This phenomenon, termed fouling, is then followed by a series of undesired reactions involving activation of complement or the coagulation cascade and adhesion of leukocytes and platelets leading to thrombus formation. Thus, considerable efforts are directed towards the preparation of fouling-resistant surfaces with the best possible hemocompatibility. Herein, a comprehensive hemocompatibility study after heparinized blood contact with seven polymer brushes prepared by surface-initiated atom transfer radical polymerization is reported. The resistance to fouling is quantified and thrombus formation and deposition of blood cellular components on the coatings are analyzed. Moreover, identification of the remaining adsorbed proteins is performed via mass spectroscopy to elucidate their influence on the surface hemocompatibility. Compared with an unmodified glass surface, the grafting of polymer brushes minimizes the adhesion of platelets and leukocytes and prevents the thrombus formation. The fouling from undiluted blood plasma is reduced by up to 99%. Most of the identified proteins are connected with the initial events of foreign body reaction towards biomaterial (coagulation cascade proteins, complement component, and inflammatory proteins). In addition, several proteins that are not previously linked with blood-biomaterial interaction are presented and discussed.

Brush-Like Interface on Surface-Attached Hydrogels Repels Proteins and Bacteria.

Interfacing artificial materials with biological tissues remains a challenge. The direct contact of their surface with the biological milieu results in multiscale interactions, in which biomacromolecules adsorb and act as transducers mediating the interactions with cells and tissues. So far, only antifouling polymer brushes have been able to conceal the surface of synthetic materials. However, their complex synthesis has precluded their translation to applications. Here, it is shown that ultrathin surface-attached hydrogel coatings of N-(2-hydroxypropyl) methacrylamide (HPMA) and carboxybetaine methacrylamide (CBMAA) provide the same level of protection as brushes. In spite of being readily applicable, these coatings prevent the fouling from whole blood plasma and provide a barrier to the adhesion of Gram positive and negative bacteria. The analysis of the components of the surface free energy and nanoindentation experiments reveals that the excellent antifouling properties stem from the strong surface hydrophilicity and the presence of a brush-like structure at the water interface. Moreover, these coatings can be functionalized to achieve antimicrobial activity while remaining stealth and non-cytotoxic to eukaryotic cells. Such level of performance is previously only achieved with brushes. Thus, it is anticipated that this readily applicable strategy is a promising route to enhance the biocompatibility of real biomedical devices.

Thermoresponsive, Pyrrolidone‐Based Antifouling Polymer Brushes

AbstractCommonly, modification of surfaces with thermoresponsive polymers is performed using poly(N‐isopropylacrylamide) (poly(NIPAM)). However, integration of poly(NIPAM) with a second polymer to obtain more complex copolymer structures has proven challenging due to inherently poorly controllable polymerization characteristics of acrylamides. In this study, (N‐(2‐methacryloyloxyethyl)pyrrolidone (NMEP) is synthesized and polymerized under controlled conditions from silicon oxide substrates via surface‐initiated atom transfer radical polymerization (SI‐ATRP) to produce thermoresponsive poly(NMEP) brushes. The livingness of the brushes is demonstrated by reinitiation of poly(NMEP) brushes using oligo(ethylene glycol) methyl ether methacrylate to obtain diblock copolymer brushes. Following extensive characterization, the reversible thermoresponsive behavior of these poly(NMEP) brushes is demonstrated using phase‐controlled AFM topography measurements in an aqueous liquid environment. These measurements indicate that at 27 °C the poly(NMEP) brushes are solvated and extend away from the surface, whereas at 60 °C the polymers are insoluble and reside in a collapsed conformation. Finally, to investigate the potential applicability of poly(NMEP) brushes in biomedical devices, the antifouling properties of the coating are tested in aqueous media containing BSA, fibrinogen, or 10% diluted human serum using quartz crystal microbalance with dissipation monitoring (QCM‐D). These measurements reveal very good antifouling properties, even when exposed to 10% diluted human serum.

Counterpropagating Gradients of Antibacterial and Antifouling Polymer Brushes.

We report on the formation of counterpropagating density gradients in poly([2-dimethylaminoethyl] methacrylate) (PDMAEMA) brushes featuring spatially varying quaternized and betainized units. Starting with PDMAEMA brushes with constant grafting density and degree of polymerization, we first generate a density gradient of quaternized units by directional vapor reaction involving methyl iodide. The unreacted DMAEMA units are then betainized through gaseous-phase betainization with 1,3-propanesultone. The gas reaction of PDMAEMA with 1,3-propanesultone eliminates the formation of byproducts present during the liquid-phase modification. We use the counterpropagating density gradients of quaternized and betainized PDMAEMA brushes in antibacterial and antifouling studies. Completely quaternized and betainized brushes exhibit antibacterial and antifouling behaviors. Samples containing 12% of quaternized and 85% of betainized units act simultaneously as antibacterial and antifouling surfaces.

A CRISPR/Cas12a-assisted on-fibre immunosensor for ultrasensitive small protein detection in complex biological samples

Tracking trace amounts of analytes directly from low volumes of complex biological samples remains an ongoing challenge in precision diagnostics, as the commonly used immunosorbent assays have limited sensitivity. Herein, a CRISPR/Cas12a assisted on-fibre immunosensor (CAFI) was developed based on an antibody-analyte-aptamer sandwich structure, in which a single strand DNA aptamer was applied to detect the analyte while triggering the CRISPR/Cas12a fluorescent detection system to amplify the analyte signal. This novel CAFI biosensing system was fabricated on a glass fibre surface with an antifouling PEG polymer brush modified for the detection of a spectrum of small molecules from complex media. In comparison with a conventional ELISA system, CAFI has a 1,000-fold higher sensitivity with the limit of detection for IFN-γ down to 1 fg mL−1 (58.8 aM). It also has a tuneable linear detection range that can be easily adjusted within the range 1 fg mL−1 to 100 pg mL−1 (5 orders of magnitude), meeting the requirements of the demanding diagnostic scenarios. CAFI has successfully been demonstrated by detecting IFN-γ from a diverse complex biological sample type, including human serum, whole blood, perspiration, and saliva. Moreover, CAFI is applicable for the detection of other analytes by simply modifying the capture antibody and detection aptamer, demonstrated here with insulin. All these superior capabilities of CAFI make it a suitable technology to measure proteins in low (100 μL) volume complex biological samples.

Unraveling the Mechanism and Kinetics of Binding of an LCI-eGFP-Polymer for Antifouling Coatings.

The ability of proteins to adsorb irreversibly onto surfaces opens new possibilities to functionalize biological interfaces. Herein, the mechanism and kinetics of adsorption of protein-polymer macromolecules with the ability to equip surfaces with antifouling properties are investigated. These macromolecules consist of the liquid chromatography peak I peptide from which antifouling polymer brushes are grafted using single electron transfer-living radical polymerization. Surface plasmon resonance spectroscopy reveals an adsorption mechanism that follows a Langmuir-type of binding with a strong binding affinity to gold. X-ray reflectivity supports this by proving that the binding occurs exclusively by the peptide. However, the lateral organization at the surface is directed by the cylindrical eGFP. The antifouling functionality of the unimolecular coatings is confirmed by contact with blood plasma. All coatings reduce the fouling from blood plasma by 8894% with only minor effect of the degree of polymerization for the studied range (DP between 101 and 932). The excellent antifouling properties, combined with the ease of polymerization and the straightforward coating procedure make this a very promising antifouling concept for a multiplicity of applications.

θ-Solvent-Mediated Double-Shell Polyethylene Glycol Brushes on Nanoparticles for Improved Stealth Properties and Delivery Efficiency.

Antifouling polymer brushes are widely used to inhibit the formation of protein corona on nanoparticles (NPs) and subsequent accumulation in the liver and spleen. Herein, we demonstrate a θ-solvent-mediated method for the preparation of gold nanoparticles with a high polyethylene glycol (PEG) grafting density. Reaching the θ-solvent by adding salt (e.g., Na2SO4) can significantly increase the grafting density of the PEG brush to 2.08 chains/nm2. The PEG polymer brush prepared in the θ-solvent possesses a double-shell structure consisting of a concentrated polymer brush (CPB) and a semidilute polymer brush (SDPB), denoted as NP@CPB@SDPB, while those prepared in a good solvent have only a SDPB shell, i.e., NP@SDPB. Compared to the NP@SDPB structure, the NP@CPB@SDPB structure decreases the liver accumulation from 34.0%ID/g to 23.1%ID/g, leading to an increase in tumor accumulation from 8.5%ID/g to 12.8%ID/g. This work provides new insights from the perspective of polymer physical chemistry into the improved stealth properties and delivery efficiency of NPs, which will accelerate the clinical translation of nanomedicine.

Machine Learning-Enabled Repurposing and Design of Antifouling Polymer Brushes

Rational development of antifouling materials is of great importance for fundamental research and real-world applications. However, current experimental designs and computational modelings of antifouling materials still retain empirical flavor due to the data complexity of polymers and their associated structures/properties. In this work, we developed a data-driven, machine learning workflow, in combination with an in-house benchmark dataset of antifouling polymer brushes, to discover the potential antifouling property of existing polymer brushes using the descriptor-based artificial neural network (ANN) model and design the new antifouling polymer brushes using the group-based supporting vector regression (SVR) model. The resultant two machine learning models not only demonstrated their reliability, predictivity, and applicability, but also established the composition-structure–property relationships using both descriptors and functional groups. Finally, we synthesized different repurposed and newly designed polymer brushes, as predicted by ANN and SVR models, all of which exhibited excellent surface resistance to protein adsorption from undiluted human blood serum and plasma at optimal film thicknesses. Overall, our data-driven machine learning models can be used as an intelligent tool for determining, repurposing, and designing new superior antifouling materials beyond polymer brushes.

Antifouling Surfaces Enabled by Surface Grafting of Highly Hydrophilic Sulfoxide Polymer Brushes.

Antifouling surfaces are important in a broad range of applications. An effective approach to antifouling surfaces is to covalently attach antifouling polymer brushes. This work reports the synthesis of a new class of antifouling polymer brushes based on highly hydrophilic sulfoxide polymers by surface-initiated photoinduced electron/energy transfer-reversible addition-fragmentation chain transfer (PET-RAFT) polymerization. The sulfoxide polymer brushes are able to effectively reduce nonspecific adsorption of proteins and cells, demonstrating remarkable antifouling properties. Given the outstanding antifouling behavior of the sulfoxide polymers and versatility of surface-initiated PET-RAFT technology, this work presents a useful and general approach to engineering various material surfaces with antifouling properties, for potential biomedical applications in areas such as tissue engineering, medical implants, and regenerative medicine.

Controlled Surface Adhesion of Macrophages via Patterned Antifouling Polymer Brushes

Macrophages will play an important role in future diagnostics and immunotherapies of cancer. However, this demands to selectively capture and sort different subpopulations, which remains a challenge due to their innate ability to bind to a wide range of interfaces indiscriminately. The main obstacle here is the lack of interfaces combining sufficient antifouling properties with the display of specific binding sites allowing sorting and quantification. Herein, as a proof of principle means, it is introduced to pattern interfaces to locally and selective capture macrophages. The repellent coating is based on antifouling polymer brushes, which can be functionalized. Arrays of binding sites are constructed by microchannel cantilever spotting. Those structures are tested for the isolation of different macrophage subtypes, especially polarized anti‐inflammatory macrophages of the M2 type which can be found associated to tumors (“tumor associated macrophages”; TAMs). Using macrophages as a model system, it is demonstrated that the newly developed surfaces and patterns are efficient for specifically trapping targeted cells and can be useful for further development of therapeutic or diagnostic purposes in the future.

Robocoliths : An Engineered Coccolith-Based Hybrid that Transforms Light into Swarming Motion

The establishment of controlled nano- and mesoscopic energized entities that gather, in a concerted effort, into motile aggregated patterns is at the forefront of scientific discovery. However, translating energy into swarming motion for such miniature-based entities remains a challenge. This requires simultaneously breaking the symmetry of the system to enable locomotion and a coupling effect between the objects that are part of the population to induce the collective motion. Herein, we introduce Robocoliths as a new concept of light-driven Emiliania huxleyi (EHUX) coccolith-based miniature entities capable of swarming behavior. EHUX coccoliths are characterized by an asymmetric morphology, which is a crucial advantage in the design of nano- and mesoscopic objects with locomotory abilities. Their activation with the bioinspired material polydopamine not only endowed the asymmetric coccoliths with advanced functionalities such as thermal and energy harvesting responsiveness under visible light exposure; it also provided a functional surface from which antifouling polymer brushes were grown. The energy harvesting responsive Robocoliths can induce an increase of temperature in the surrounding environment while displaying a collective behavior (i.e., swarming) via a controlled ON-OFF light switching mechanism. In this context, Robocoliths could pave the way for a new generation of multifunctional swarming bio-micromachines with a potential impact in various fields of applications, starting from the analytical and environmental sectors and extending to the therapeutically relevant domains.

Surface plasmon resonance-based aptasensor for direct monitoring of thrombin in a minimally processed human blood

Optical affinity biosensors are pursued for timely monitoring of thrombin in human blood, which is of urgent need in tailored anticoagulation therapies. However, the unspecific deposition of molecules, cells, and aggregates from the blood at their surface (also termed fouling) severely hinders their development and impedes the deploying of this technology to everyday clinical practice. We addressed this challenge by designing surface plasmon resonance (SPR) sensor chip with an antifouling polymer brush architecture and incorporated thrombin aptamer bioreceptors. Poly[(N-(2-hydroxypropyl)-methacrylamide)-co-(carboxybetaine methacrylamide)] brushes were synthesized on gold sensor chip surface via photoinduced single-electron transfer living radical polymerization and postmodified with three thrombin aptamers (HD1 short, HD1 and HD22). The affinity interaction of the aptamer bioreceptors with thrombin (as well as with other molecules present in the blood) was investigated and changes in their performance when incorporated into the polymer brushes were characterized. The combination of brushes and aptamer bioreceptors allowed for the analysis of medically relevant concentrations of thrombin in the 10 % blood by direct SPR detection format. This is the first time that the optical affinity biosensor is demonstrated for label-free analysis of biomarkers in a minimally processed human blood without a need for pre-separation steps. We believe that this system constitutes a basis for the future affinity biosensor applications that are suitable for the clinical settings and can be readily adapted to detect a range of important biological markers.

C1q recognizes antigen-bound IgG in a curvature-dependent manner

C1q is an important recognition protein in the complement system, which is a major protein cascade in the innate immune system. Upon recognition of a target by C1q, the target is marked for opsonization and destruction. C1q recognizes many pathogenic patterns directly, but an important target is the Fc domain of antibodies binding to their antigen. In this paper, the curvature-dependence of the interaction between IgG and C1q is studied by surface plasmon resonance and quartz crystal microbalance. IgG is organized in similar surface coverage densities on flat polystyrene surfaces and polystyrene nanoparticles of different sizes, and the amount of C1q binding to the IgG is investigated. Nanoparticles in solution were found to aggregate upon incubation with IgG, and therefore a new technique utilizing nanoparticles binding to antifouling polymer brush functionalized surfaces was used to prepare surfaces with nanoparticles for measurements with surface plasmon resonance. Interestingly antigen-bound IgG at the curved surface of nanoparticles showed 5.6 times lower binding of C1q compared to at matched flat surfaces. There was no significant difference between the binding at 100 and 200 nm polystyrene particles. These findings are important for designing drug delivery systems to evade the complement system.