Yolk-Shell Encapsulation of Cells by Biomimetic Mineralization and Visible Light-Induced Surface Graft Polymerization.

The pursuit of low-cytotoxicity modification strategies represents a prominent avenue in cell coating research, holding immense significance for the advancement of practical living cell-related technologies. Here, we presented a novel method to fabricate encapsulated yeast cells with a yolk-shell structure by biomimetic mineralization and visible-light-induced surface graft polymerization. In this approach, an amorphous calcium carbonate (ACC) shell was first deposited on the surface of a yeast cell (cell@ACC) modified with 4 layers of self-assembled poly(diallyl dimethylammonium chloride) (PDADMAC)/poly(acrylic acid) (PAA) film using a biomimetic mineralization technique. Subsequently, polyethylenimine (PEI) was absorbed on the surface of cell@ACC by electrostatic interaction. Then, a cross-linked shell was introduced by surface-initiated graft polymerization of poly(ethylene glycol) diacrylate (PEGDA) on cell@ACC under irradiation of visible light using thioxanthone catechol-O,O'-diacetic acid as the photosensitizer. After the removal of the inner ACC shell, the yolk-shell-structured yeast cells (cell@PHS) were obtained. Due to the mild conditions of the strategy, the cell@PHS could retain 98.81% of its original viability. The introduction of the shell layer significantly prolonged the lag phase of yeast cells, which could be tuned between 5 and 25 h by regulating the thickness of the shell. Moreover, the cell@PHS showed improved resistance against lyticase due to the presence of a protective shell. After 30 days of storage, the viability of cell@PHS was 81.09%, which is significantly higher than the 19.89% viability of native yeast cells.

Self-Cross-Linkable Maleic Anhydride Terpolymer Coating with Inherent High Antimicrobial Activity and Low Cytotoxicity.

Developing coating materials with low cytotoxicity and high antimicrobial activity has been recognized as an effective way to prevent medical device-associated infections. In this study, a maleic anhydride terpolymer (PPTM) is synthesized and covalently attached to silicone rubber (SR) surface. The formed coating can be further cross-linked (SPM) through the self-condensation of pendent siloxane groups of terpolymer. No crack or delamination of SPM was observed after 500 cycles of bending and 7 day immersion in deionized water. The sliding friction force of a catheter was reduced by 50% after coating with SPM. The SPM coating without adding any extra antibacterial reagents can kill 99.99% of Staphylococcus aureus and Escherichia coli and also significantly reduce bacterial coverage, while the coating displayed no antimicrobial activity when maleic anhydride groups of SPM were aminated or hydrolyzed. The results of the repeated disinfection tests showed that the SR coated with SPM could maintain 87.3% bactericidal activity within 5 cycles. Furthermore, the SPM coating only imparted slight toxic effect (>85% viability) on L929 cells after 36 h of coculture, which is superior to the coating of aminated SPM conjugated with the antimicrobial peptide E6. The terpolymer containing maleic anhydride units have great potential as a flexible and durable coating against implant infections.

UV-Induced Thiol-Ene "Click" Surface Grafting Polymerization on BOPP Substrate and Its Postmodifying for Hydrophilic and Antibacterial Applications.

This paper proposed a novel and versatile surface modification route by integrating UV light-mediated thiol-ene "click" surface grafting polymerization and postmodification via the reactions of the surface thiol groups. At first, poly(thiol ether) layers with tunable thiol group density, up to 8.2 × 102 ea/nm3 for cross-linked grafting layers, were grafted from biaxially oriented polypropylene (BOPP) film. Then, the surface -SH groups reacted with epoxy compounds to introduce quaternary ammonium salt. With the immobilized quaternary ammonium salt and coordinated Zn2+ ions, the modified film demonstrated 99.98% antibacterial rate against Staphylococcus aureusafter soaking in DI water for 21 days and in a highly alkaline environment (0.1 M NaOH aqueous solution) for 3 days, and the surface water contact angle decreased to 39°. At last, the polymethacrylate chains were also successfully grafted from the surface thiol groups of the cross-linked poly(thiol ether) under visible light irradiation. With 2-(dimethyldodecylammonium) ethyl methacrylate as the grafting monomer, the modified BOPP film had shown a 99.99% antibacterial rate against both Escherichia coliand S. aureus. Meanwhile, with 2-methacryloxyethyl phosphoryl choline as grafting monomer, the modified surface showed an excellent antibioadhesion of living S. aureus, and the surface water contact angle was as low as 48°.

A Novel Family of Stable Polyelectrolyte Complexes Based on Mixed Olefins-Maleic Anhydride Copolymer.

In the present study, the copolymer of mixed olefins included in unetherified gasoline and maleic anhydride (PUGM) was prepared by self-stabilized precipitation polymerization (2SP) and employed for the synthesis of a new family of stable polyelectrolyte complexes (PECs). Polyanionic saponified PUGM partially grafted with methoxy poly(ethylene glycol) (PUGMS-g-mPEG) and polycationic quaternized PUGM (PUGMQ) were both derived from PUGM via the facile modification of anhydride groups. The particle size, zeta potential, morphology, and stability of self-assembled PEC particles were investigated thoroughly. Strikingly, the introduction of long mPEG side chains (Mn = 4000) had a remarkable effect on the self-assembled particles, which displayed a constant particle size of ∼200 nm regardless of varying n+/n-. Moreover, it also enhanced the salt tolerance and long-term stability of PEC particles significantly. Our work not only provides an effective approach to PECs from petroleum resources with low cost but also deepens the understanding of the relationship between the chain structure of polyelectrolytes and the stability of PECs.

Super-adsorbent microspheres based on a triallyl isocyanurate-maleic anhydride copolymer for the removal of organic pollutants from water.

Owing to the frequent occurrence of diclofenac sodium (DS) in fresh aquatic environments and its potential toxicity towards living organisms, the effective removal of DS has attracted worldwide attention. Herein, a green and efficient strategy to fabricate crosslinked microspheres with interconnected mesoporous structures and abundant adsorption active sites was developed. With this strategy, triallyl isocyanurate (TAIC)-maleic anhydride (MAH) copolymer microspheres (TMs) with a diameter of 1.19-1.35 µm were first prepared by self-stabilized precipitation (2SP) polymerization, and the TMs possess a large amount reactive anhydride groups (62.5-71.8 mol%), a specific surface area of 51.6-182.4 m2 g-1 and a mesoporous structure (average pore size: 3.4-3.8 nm). Then the TMs were further functionalized with polyethylenimine (PEI) to give rise to cationic microspheres (Cat-TMs), which showed excellent adsorption performance to DS with a rapid adsorption rate (reached equilibrium within 30 min), a very high equilibrium adsorption capacity (1421 mg g-1) and excellent recyclability. The pseudo-second-order model and Langmuir model were a good fit for the adsorption kinetic and isotherm process, respectively. Furthermore, due to the high cation density (4.291 mmol g-1) and excellent pH buffer capacity of Cat-TMs, the adsorption capacity can be maintained at a high level within the pH range of 6-10. The regenerated Cat-TMs showed only a slight loss (<5%) in the adsorption capacity even after 5 adsorption-desorption cycles. In short, Cat-TMs can be considered as a highly promising adsorbent for the rapid and ultra-efficient removal of anionic organic contaminants and have significant potential to be applied in wastewater treatment.

Covalently Attaching Hollow Silica Nanoparticles on a COC Surface for the Fabrication of a Three-Dimensional Protein Microarray.

Compared to traditional two-dimensional (2D) biochips, three-dimensional (3D) biochips exhibit the advantages of higher probe density and detection sensitivity due to their designable surface microstructure as well as enlarged surface area. In the study, we proposed an approach to prepare a 3D protein chip by deposition of a monolayer of functionalized hollow silica nanoparticles (HSNs) on an activated cyclic olefin copolymer (COC) substrate. First, the COC substrate was chemically modified through the photografting technique to tether poly[3-(trimethoxysilyl) propyl methacrylate] (PTMSPMA) brushes on it. Then, a monolayer of HSNs was deposited on the modified COC and covalently attached via a condensation reaction between the hydrolyzed pendant siloxane groups of PTMSPMA and the Si-OH groups of HSNs. The roughness of the COC substrate significantly increased to 50.3 nm after depositing a monolayer of HSNs (ranging from 100 to 700 nm), while it only caused a negligible reduction in the light transmittance of COC. The HSN-modified COC was further functionalized with epoxide groups by a silane coupling agent for binding proteins. Immunoglobulin G could be effectively immobilized on this substrate with the highest immobilization efficiency of 75.2% and a maximum immobilization density of 1.236 µg/cm2, while the highest immobilization efficiency on a 2D epoxide group-modified glass slide was only 57.4%. Moreover, immunoassay results confirmed a competitive limit of detection (LOD) (1.06 ng/mL) and a linear detection range (1-100 ng/mL) of the 3D protein chip. This facile and effective approach for fabricating nanoparticle-based 3D protein microarrays has great potential in the field of biorelated detection.

Three-dimensional protein microarrays fabricated on reactive microsphere modified COC substrates.

Fabrication of three-dimensional (3D) surface structures for the high density immobilization of biomolecules is an effective way to prepare highly sensitive biochips. In this work, a strategy to attach polymeric microspheres on a cyclic olefin copolymer (COC) substrate for the preparation of a 3D protein chip was developed. The COC surface was firstly functionalized by the photograft technique with epoxy groups, which were subsequently converted to amine groups. Then monodisperse poly(styrene-alt-maleic anhydride) (PSM) copolymer microspheres were prepared by self-stabilized precipitation polymerization and deposited as a single layer on a modified COC surface to form a 3D surface texture. The surface roughness of the COC support undergoes a significant increase from 1.4 nm to 37.1 nm after deposition of PSM microspheres with a size of 460 nm, and the modified COC still maintains a transmittance of more than 63% at the fluorescence excitation wavelengths (555 nm and 647 nm). The immobilization efficiency of immunoglobulin G (IgG) on the 3D surface reached 75.6% and the immobilization density was calculated to be 0.255 µg cm-2, at a probe protein concentration of 200 µg mL-1. The 3D protein microarray can be rapidly blocked by gaseous ethylenediamine within 10 minutes due to the high reactivity of anhydride groups in PSM microspheres. Immunoassay results show that the 3D protein microarray achieved specific identification of the target protein with a linear detection range from 6.25 ng mL-1 to 250 ng mL-1 (R2 > 0.99) and a limit of detection of 8.87 ng mL-1. This strategy offers a novel way to develop high performance polymer-based 3D protein chips.

A Versatile Strategy to Coat Individual Cell with Fully/Partially Covered Shell for Preparation of Self-Propelling Living Cells.

Coating living cells with a functional shell has been regarded as an effective way to protect them against environmental stress, regulate their biological behaviors, or extend their functionalities. Here, we reported a facile method to prepare fully or partially coated shells on an individual yeast cell surface by visible light-induced graft polymerization. In this strategy, yeast cells that were surface-absorbed with polyethylenimine (PEI) were deposited on the negatively charged glass slide to form a single layer by electrostatic interaction. Then, surface-initiated graft polymerization of poly(ethylene glycol) diacrylate (PEGDA) on yeast cells under visible light irradiation was carried out to generate cross-linked shells on the cells. The process of surface modification had negligible influence on the viability of yeast cells due to the mild reaction condition. Additionally, compared to the native yeast cells, a 17.5 h of delay in division was observed when the graft polymerization was performed under 15 mW/cm2 irradiation for 30 min. Introducing artificial shell endowed yeast cells with significant resistance against lyticase, and the protection can be enhanced by increasing the thickness of shell. Moreover, the partially coated yeast cells would be prepared by simply adjusting the reaction condition such as irradiation density and time. By immobilizing urease on the functional patch, the asymmetrically modified yeast cells exhibited self-propelling capability, and the speed of directional movement reached 4 µm/s in the presence of 200 mM urea. This tunable coating individual cell strategy with varying functionality has great potential applications in fields of cell-based drug delivery, cell therapy, biocatalysis, and tissue engineering.

UV-Assisted Li+-Catalyzed Radical Grafting Polymerization of Vinyl Ethers: A New Strategy for Creating Hydrolysis-Resistant and Long-Lived Polymer Brushes as a "Smart" Surface Coating.

A facile synthetic route was developed to prepare a surface-grafted brush layer of poly(vinyl ethers) (PVEs) directly by a radical mechanism, with the "naked" Li+ acting as a catalyst. Density functional theory calculations suggested that complexation of naked Li+ to VEs significantly reduced the highest unoccupied molecular orbital-lowest unoccupied molecular orbital (HOMO-LUMO) energy gap from 5.08 to 0.68 eV, providing a better prospect for electron transfer. The structure, morphology, and surface properties of grafted polymer layers were characterized using attenuated total reflection Fourier transform infrared spectroscopy, Raman spectroscopy, X-ray photoelectron spectroscopy, atomic force microscopy, and dynamic water contact angle (DCA). Moreover, ellipsometry data indicated that the thickness of the polymer brushes was in the range of 20-60 nm, which corresponds to the grafting densities of 0.65-1.15 chain/nm2, and DCA decreased from 84.4 to 45.3°. Most importantly, no hydrolysis was observed for the modified surface after 30 days of exposure to phosphate-buffered saline solution, 0.1 mol/L NaOH(eq) and 0.1 mol/L HCl(eq), demonstrating excellent hydrolysis resistance with long service life. In addition, as a proof of concept, the side hydroxyl groups of grafted PVEs provide active sites for efficient fixation of bioactive molecules, e.g., glycosaminoglycan and serum protein.

Antibacterial hydrogel coating: Strategies in surface chemistry.

Hydrogels have emerged as promising antimicrobial materials due to their unique three-dimensional structure, which provides sufficient capacity to accommodate various materials, including small molecules, polymers and particles. Coating substrates with antibacterial hydrogel layers has been recognized as an effective strategy to combat bacterial colonization. To prevent possible delamination of hydrogel coatings from substrates, it is crucial to attach hydrogel layers via stronger links, such as covalent bonds. To date, various surface chemical strategies have been developed to introduce hydrogel coatings on different substrates. In this review, we first give a brief introduction of the major strategies for designing antibacterial coatings. Then, we summarize the chemical methods used to fix the antibacterial hydrogel layer on the substrate, which include surface-initiated graft crosslinking polymerization, anchoring the hydrogel layer on the surface during crosslinking, and chemical crosslinking of layer-by-layer coating. The reaction mechanisms of each method and matched pretreatment strategies are systemically documented with the aim of introducing available protocols to researchers in related fields for designing hydrogel-coated antibacterial surfaces.

Facile fabrication of shell crosslinked microcapsule by visible light induced graft polymerization for enzyme encapsulation.

Immobilization of enzymes can effectively improve their stability, facilitate their recycling and reduce the cost, which is of great significance for the development of highly efficient biocatalysis technology. Here a simple strategy to encapsulate enzymes into polymeric microcapsules fabricated by visible light induced graft polymerization on a removable template was developed. The strategy showed a high degree of enzyme loading and excellent reusability of the immobilized enzyme.

Facile Surface Functionalization of Cyclic Olefin Copolymer Film with Anhydride Groups for Protein Microarray Fabrication.

Immobilization of protein at high efficiency is a challenge for fabricating polymer-based protein chips. Here, a simple but effective approach was developed to fabricate a cyclic olefin copolymer (COC)-based protein microarray with a high immobilization density. In this strategy, poly(maleic anhydride-co-vinyl acetate) (poly(MAH-co-VAc)) brushes were facilely attached on the COC surface via UV-induced graft copolymerization. The introduction of poly(MAH-co-VAc) brushes resulted in an obvious increase in the surface roughness of COC. The functionalized COC showed little reduction in transparency compared with pristine COC, indicating that the photografting treatment did not alter its optical property. The graft density of the anhydride groups on the modified COC could be tuned from 0.46 to 3.2 µmol/cm2. The immobilization efficiency of immunoglobulin G (IgG) on functionalized COC reached 88% due to the high reactivity between anhydride groups and amine groups of IgGs. An immunoassay experiment demonstrated that the microarray showed high sensitivity to the target analyte.

Layered Co-Immobilization of ß-Glucosidase and Cellulase on Polymer Film by Visible-Light-Induced Graft Polymerization.

Exploring a suitable immobilization strategy to improve catalytic efficiency and reusability of cellulase is of great importance to lowering the cost and promoting the industrialization of cellulose-derived bioethanol. In this work, a layered structure with a thin PEG hydrogel as the inner layer and sodium polyacrylate (PAANa) brush as the outer layer was fabricated on low density polyethylene (LDPE) film by visible-light-induced graft polymerization. Two enzymes, ß-glucosidase (BG) and cellulase, were separately coimmobilized onto this hierarchical film. As supplementary to cellulase for improving catalytic efficiency, BG was in situ entrapped into the inner PEG hydrogel layer during the graft polymerization from the LDPE surface. After graft polymerization of sodium acrylate on the PEG hydrogel layer was reinitiated, cellulase was covalently attached on the outer PAANa brush layer. Owing to the mild reaction condition (visible-light irradiation and room temperature), the immobilized BG could retain a high activity after the graft polymerization. The immobilization did not alter the optimal pH and temperature of BG or the optimal temperature of cellulase. However, the optimal pH of cellulase shifts to 5.0 after immobilization. Compared with the original activity of single cellulase system and isolated BG/cellulase immobilization system, the dual-enzyme system exhibited 82% and 20% increase in catalytic activity, respectively. The dual-enzyme system could maintain 93% of carboxymethylcellulose sodium salt (CMC) activity after repeating 10 cycles of hydrolysis and 89% of filter paper activity after 6 cycles relative to original activity, exhibiting excellent reusability. This layer coimmobilization system of BG and cellulase on the polymer film displays tremendous potential for practical application in a biorefinery.

Highly Transparent Cyclic Olefin Copolymer Film with a Nanotextured Surface Prepared by One-Step Photografting for High-Density DNA Immobilization.

Compared with conventional glass slides and two-dimensional (2D) planar microarrays, polymer-based support materials and three-dimensional (3D) surface structures have attracted increasing attention in the field of biochips because of their good processability in microfabrication and low cost in mass production, as well as their improved sensitivity and specificity for the detection of biomolecules. In the present study, UV-induced emulsion graft polymerization was carried out on a cyclic olefin copolymer (COC) surface to generate 3D nanotextures composed of loosely stacked nanoparticles with a diameter of approximately 50 nm. The introduction of a hierarchical nanostructure on a COC surface only resulted in a 5% decrease in its transparency at a wavelength of 550 nm but significantly increased the surface area, which markedly improved immobilization density and efficiency of an oligonucleotide probe compared with the functional group and polymer brush-modified substrates. The highest immobilization efficiency of the probes reached 93%, and a limit of detection of 75 pM could be obtained. The hybridization experiment demonstrated that the 3D gene chip exhibited excellent sensitivity for target DNA detection and single-nucleotide polymorphism discrimination. This one-step approach to the construction of nanotextured surfaces on the COC has promising applications in the fields of biochips and immunoassays.

Cytocompatible Fabrication of Yeast Cells/Fabrics Composite Sheet for Bioethanol Production.

Entrapment of living cells into a polymer network has significant potential in various fields such as biomass conversion and tissue engineering. A crucial challenge for this strategy is to provide a mild enough condition to preserve cell viability. Here, a facile and cytocompatible method to entrap living yeast cells into a poly(ethylene glycol) (PEG) network grafting from polypropylene nonwoven fabrics via visible-light-induced surface living graft crosslinking polymerization is reported. Due to the mild reaction conditions and excellent biocompatibility of PEG, the immobilized yeast cells could maintain their viability and proliferate well. The obtained composite sheet has excellent long-term stability and shows no significant efficiency loss after 25 cycles of repeated batch bioethanol fermentation. The immobilized yeast cells exhibit 18.0% higher bioethanol fermentation efficiency than free cells. This strategy for immobilization of living cells with high viability has significant potential application.

Antibacterial Hydrogels.

Antibacterial materials are recognized as important biomaterials due to their effective inhibition of bacterial infections. Hydrogels are 3D polymer networks crosslinked by either physical interactions or covalent bonds. Currently, hydrogels with an antibacterial function are a main focus in biomedical research. Many advanced antibacterial hydrogels are developed, each possessing unique qualities, namely high water swellability, high oxygen permeability, improved biocompatibility, ease of loading and releasing drugs, and structural diversity. Here, an overview of the structures, performances, mechanisms of action, loading and release behaviors, and applications of various antibacterial hydrogel formulations is provided. Furthermore, the prospects in biomedical research and clinical applications are predicted.

Decorating an individual living cell with a shell of controllable thickness by cytocompatible surface initiated graft polymerization.

Surface engineering of individual living cells is a promising field for cell-based applications. However, engineering individual cells with controllable thickness by chemical methods has been rarely studied. This article describes the development of a new cytocompatible chemical strategy to decorate individual living cells. The thicknesses of the crosslinked shells could be conveniently controlled by the irradiation time, visible light intensity, or monomer concentration. Moreover, the lag phase of the yeast cell division was extended and their stability against lysis was improved, which could also be tuned by controlling the shell thickness.

Separated Immobilization of Incompatible Enzymes on Polymer Substrate via Visible Light Induced Living Photografting Polymerization.

The use of the mixed catalytic system with several enzymes can provide multiple benefits in terms of the cost, simplification of a multistep reaction, and effectiveness of complex chemical reactions. Although study of different enzyme coimmobilization systems has attracted increasing attention in recent years, separately immobilizing enzymes which can not coexist on one support is still one of the great challenges. In this paper, a simple and effective strategy was introduced to separately encapsulate incompatible trypsin and transglutaminase (TGase) into different poly(ethylene glycol) (PEG) network layer grafted on low-density polyethylene (LDPE) film via visible light induced living photografting polymerization. As a proof of concept, this dual-enzyme separately loaded film was used to catalyze the synthesis of a new target antitumor drug LTV-azacytidine. The final results demonstrated that this strategy could maintain higher activities of both enzymes than the mixed coimmobilization method. And the mass spectra analysis results demonstrated that LTV-azacytidine was successfully synthesized. We believe that this facile and mild separately immobilizing incompatible enzyme strategy has great application potential in the field of biocatalysis.

Hierarchical PEG-Based 3D Patterns Grafting from Polymer Substrate by Surface Initiated Visible Light Photolithography.

The precise construction of a hierarchical complex pattern on substrates is required for numerous applications. Here, a strategy to fabricate well-defined hierarchical three dimensional (3D) patterns on polymer substrate is developed. This technique, which combines photolithography and visible light-induced surface initiated living graft crosslinking polymerization (VSLGCP), can effectively graft 3D patterns onto polymer substrate with high fidelity and controllable height. Owing to the living nature of VSLGCP, hierarchical 3D patterns can be prepared when a sequential living graft crosslinking process is performed on the first formed patterns. As a proof-of-concept, a reactive two layer 3D pattern with a morphology of lateral stripe on vertical stripe is prepared and employed to separately immobilize model biomolecules, e.g., biotin and IgG. This two component pattern can specifically interact with corresponding target proteins successfully, indicating that this strategy has potential applications in the fabrication of polymer-based multicomponent biomolecule microarrays.

Ammonium-Functionalized Hollow Polymer Particles As a pH-Responsive Adsorbent for Selective Removal of Acid Dye.

In this work, a novel type of ammonium-functionalized hollow polymer particles (HPP-NH3(+)) with a high density of ammonium groups in the shell has been specially designed and synthesized. Benefiting from both the high surface area and from the high density of positively charged ammonium groups, the as-prepared HPP-NH3(+) can serve as a selective adsorbent for the removal of negatively charged acid dye (e.g., methyl blue a-MB). The equilibrium adsorption data of a-MB on the HPP-NH3(+) were evaluated using Freundlich and Langmuir isotherm models, and Langmuir isotherm exhibited a better fit with a maximum adsorption capacity of 406 mg/g. Most importantly, because of the presence of dual functional groups (ammonium and carboxyl groups), the HPP-NH3(+) showed a significant pH-dependent equilibrium adsorption capacity, which increased dramatically from 59 mg/g to 449 mg/g as the solution pH decreased from 9 to 2. This uniqueness makes the dye-adsorbed HPP-NH3(+) can be facilely regenerated under mild condition (in weak alkaline solution, pH 10) to recover both a-MB and the HPP-NH3(+), whereas the recovery of conventional adsorbents is commonly performed under particularly severe conditions. The regenerated HPP-NH3(+) can be reused for dye removal and the dye removal efficiency remained above 98% even after five adsorption-desorption cycles. Because of its high adsorption capacity, pH-sensitivity, easy regeneration, and good reusability, the HPP-NH3(+) has great potential for the application in the field of water treatment, controlled drug release, and pH-responsive delivery.