Metal-Free Atom Transfer Radical Polymerization to Prepare Recylable Micro-Adjuvants for Dendritic Cell Vaccine.

In the development of dendritic cell (DC) vaccines, the maturation of DCs is a critical stage. Adjuvants play a pivotal role in the maturation of DCs, with a major concern being to ensure both efficacy and safety. This study introduces an innovative approach that combines high efficacy with safety through the synthesis of micro-adjuvants grafted with copolymers of 2-(methacrylamido) glucopyranose (MAG) and methacryloxyethyl trimethyl ammonium chloride (DMC). The utilization of metal-free surface-initiated atom transfer radical polymerization enables the production of safe and recyclable adjuvants. These micrometer-sized adjuvants surpass the optimal size range for cellular endocytosis, enabling the retrieval and reuse of them during the ex vivo maturation process, mitigating potential toxicity concerns associated with the endocytosis of non-metabolized nanoparticles. Additionally, the adjuvants exhibit a "micro-ligand-mediated maturation enhancement" effect for DC maturation. This effect is influenced by the shape of the particle, as evidenced by the distinct promotion effects of rod-like and spherical micro-adjuvants with comparable sizes. Furthermore, the porous structure of the adjuvants enables them to function as cargo-carrying "micro-shuttles", releasing antigens upon binding to DCs to facilitate efficient antigen delivery.

Switchable RAFT Polymerization Employing Photoresponsive HABI as a Mediator.

Recently, considerable interest has been devoted to developing switchable reversible addition fragmentation chain transfer (RAFT) polymerizations via photoactivation methods. Herein, a photo-deactivation strategy is introduced to regulate RAFT polymerization using photoresponsive hexaarylbiimidozole (HABI) as a mediator, which leads to switchable RAFT polymerization by repeated ON/OFF experiments. In comparison with well-known PET-RAFT polymerization, photo-deactivation RAFT (PD-RAFT) polymerization can be temporally stopped with UV light ON, where photoresponsive HABI can reversibly quench propagating radicals, resulting in switchable RAFT polymerization. The proposed mechanism of PD-RAFT polymerization in the presence of HABI involving radical quenching is based on ESR, NMR, GPC, MALDI-TOF-MS, and kinetics studies.

Surface-Initiated Synthesis of Cell-Specific Glycopolymers Using Live Mammalian Cells as Templates.

Molecular recognition is an important process in life activities where specificity is the key. However, the method to gain specificity are often complex and time-consuming. Herein, a novel, versatile, and effective way is developed to obtain cell-specific glycosurfaces by surface-initiated Cu-mediated reversible deactivation radical polymerization (Cu-RDRP) in an open to air fashion. Mammalian cells are used for the first time as live templates to realize cell-sugar monomer-aptation-polymerization which can produce cell-specific glycosurfaces. Both epithelial cell adhesion molecule (EpCAM) positive cells L929 and EpCAM negative cells Hela as models are used to acquire two cell-specific glycosurfaces, which can distinguish template-cells from others. The strategy is effective and convenient without the need of fixative pretreatment of cells. It is found that the specific capture does not rely on EpCAM antibodies, and the specificity is related to the composition and chain sequence of the glycopolymer brushes rather than surface morphology. In addition, these glycosurfaces keep the ability to identify the target cells after ten regenerative treatments, which provides another advantage for practical applications.

Glycopolymers for Antibacterial and Antiviral Applications.

Diseases induced by bacterial and viral infections are common occurrences in our daily life, and the main prevention and treatment strategies are vaccination and taking antibacterial/antiviral drugs. However, vaccines can only be used for specific viral infections, and the abuse of antibacterial/antiviral drugs will create multi-drug-resistant bacteria and viruses. Therefore, it is necessary to develop more targeted prevention and treatment methods against bacteria and viruses. Proteins on the surface of bacteria and viruses can specifically bind to sugar, so glycopolymers can be used as potential antibacterial and antiviral drugs. In this review, the research of glycopolymers for bacterial/viral detection/inhibition and antibacterial/antiviral applications in recent years are summarized.

Synthetic Sugar-Only Polymers with Double-Shoulder Task: Bioactivity and Imaging.

The search for novel fluorescent materials has attracted the attention of many researchers. Numerous bioimaging materials based on the aggregation-induced emission (AIE) units have been surging and could be employed in wide areas during the past two decades. In recent few years, the appearance of nonconventional fluorescence emitters without aromatic conjugated structures provides another bioimaging candidate which has the advantage of enhanced biodegradability and relatively low cost, and their luminescent mechanism can be explained by clustering-triggered emission (CTE) like AIE. In our contribution, we utilize nonaromatic sugar as a monomer to prepare a series of glycopolymers with designed components through sunlight-induced reversible addition fragmentation chain transfer polymerization; these glycopolymers can be employed in bioimaging fields due to the bioactivity coming from sugar and CTE capacity.

Thermo-resistance of ESKAPE-panel pathogens, eradication and growth prevention of an infectious biofilm by photothermal, polydopamine-nanoparticles in vitro.

Nanotechnology offers many novel infection-control strategies that may help prevent and treat antimicrobial-resistant bacterial infections. Here, we synthesized polydopamine, photothermal-nanoparticles (PDA-NPs) without further surface-functionalization to evaluate their potential with respect to biofilm-control. Most ESKAPE-panel pathogens in suspension with photothermal-nanoparticles showed three- to four-log-unit reductions upon Near-Infra-Red (NIR)-irradiation, but for enterococci only less than two-log unit reduction was observed. Exposure of existing Staphylococcus aureus biofilms to photothermal-nanoparticles followed by NIR-irradiation did not significantly kill biofilm-inhabitants. This indicates that the biofilm mode of growth poses a barrier to penetration of photothermal-nanoparticles, yielding dissipation of heat to the biofilm-surrounding rather than in its interior. Staphylococcal biofilm-growth in the presence of photothermal-nanoparticles could be significantly prevented after NIR-irradiation because PDA-NPs were incorporated in the biofilm and heat dissipated inside it. Thus, unmodified photothermal nanoparticles have potential for prophylactic infection-control, but data also constitute a warning for possible development of thermo-resistance in infectious pathogens.

Highly Branched Gradient Glycopolymer: Enzyme-Assisted Synthesis and Enhanced Bacteria-Binding Ability.

A one-pot strategy was applied to synchronize enzymatic monomer transformation with reversible addition fragmentation chain transfer (RAFT) polymerization for the synthesis of glycopolymers with highly branched gradient architectures. Also, the linear analogues, block glycopolymers, and gradient glycopolymers were also synthesized for comparison. The binding ability of glycopolymers toward bacteria was then studied by optical density (OD) test, confocal laser scanning microscopy (CLSM), and quartz crystal microbalance with dissipation (QCM-D). The results show that the highly branched gradient glycopolymers have the most remarkable bacteria-binding ability compared with the two linear analogues, gradient glycopolymers, and block glycopolymers. The highly branched glycopolymers were further used as inhibitors in the anti-infection test, demonstrating a significant inhibitory effect on preventing bacteria from infecting the cells.

Using One Photoredox Catalyst to Simultaneously Mediate Two Different Polymerizations for Photo-Triggered Multi-Component Orthogonal Polymerizations.

The development of multi-component orthogonal polymerizations (MOPs) with simple procedure and high efficiency is crucial for rational polymer synthesis. In this work, photo-triggered Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) polymerization are first investigated, employing copper(II) thioxanthone carboxylate (Cu(TX)2 ) as photoredox catalyst and sunlight as stimuli. Then, the novel MOPs using one photoredox catalyst, Cu(TX)2 to simultaneously mediate two different photopolymerizations, are successfully realized in one-pot, where photo-induced electron transfer atom transfer radical polymerization and photo-activated CuAAC polymerization can effectively proceed in a one-pot and non-interfering fashion to afford predetermined copolymers with well-defined structure.

Ultralow Crosslinked Microgel Brings Ultrahigh Catalytic Efficiency.

Microgel nanoreactors maintain the stability of metallic nanoparticles and regulate their catalytic activity. However, limited by the synthetic method, the recycling ability and long-lasting stability of microgel nanoreactors are challenged. Herein, a brand-new nanoparticle carrier, ultralow crosslinked poly(N-isopropylacrylamide-b-methacrylic acid) (P(NIPAm-b-MAA)) microgel, is synthesized based on the reversible addition-fragmentation chain transfer polymerization method and the self-crosslinking mechanism of PNIPAm. This carrier enables the easy preparation, low cost, long-lasting stability, and high catalytic efficiency of nanoreactors. As far as it is known, the catalytic reduction rates of several dye models used in this work are the highest ones in similar systems. In addition, the presence of the MAA block leads to the agglomeration and dispersion of the microgels under different pH conditions, thus realizing rapid recycling of the nanoreactors. This novel carrier has great potential for a wide range of applications in catalysis.

Ultralow Self-Cross-Linked Poly(N-isopropylacrylamide) Microgels Prepared by Solvent Exchange.

We found that the poly(N-isopropylacrylamide) (PNIPAm) synthesized by free-radical polymerization in organic phase could also form stable microgels in water through solvent exchange without chemical cross-linkers. Dynamic light scattering and transmission electron microscopy showed the larger swelling ratio and higher deformability of these microgels. Nuclear magnetic resonance and infrared spectroscopy indicated that the self-cross-linking structures in these microgels were attributed to the hydrogen atom abstraction both from the isopropyl tert-carbon atoms and the vinyl tert-carbon atoms in PNIPAm chains and the organic solvents were important assistants in the hydrogen abstraction behavior. Our discovery revealed that the self-cross-linking of PNIPAm chains is a common phenomenon within their free-radical polymerization process, whether in aqueous phase or in organic phase. Besides, the addition of second monomers will not affect the cross-linkage of the PNIPAm portion, which may be of great significance for the synthesis of various functional ultralow cross-linking PNIPAm microgels.

One-Pot, Multicomponent Strategy for Designing Lymphoseek-Inspired Hetero-Glycoadjuvant@AuNPs.

Searching for vaccines (antigen and adjuvant) with easy preparation and strong T-cell response are crucial for antitumor immunity. In this work, to design lymphoseek-inspired vaccine possessing the abilities of promoting vaccine internalization and enhancing CD8+ T-cell responses, a simple multicomponent strategy is successfully utilized to fabricate lymph node and dendritic cell dual-targeting glycoadjuvant@AuNPs in one pot, where three different components, catechol-containing glycopolymer, HAuCl4 , and amine-terminal CpG (CpG-NH2 ) can react in a single step to generate target adjuvant. It is found that hetero-glycoadjuvant@AuNPs could increase adjuvant internalization and enhance the activation of bone-marrow-derived dendritic cells. Critically, lymphoseek-inspired vaccine potentiates antigen-specific CD8+ T-cell immune responses.

Topological Glycopolymers as Agglutinator and Inhibitor: Cyclic versus Linear.

Carbohydrates play an important role in biological processes for their specific interactions with proteins. Cyclic glycopolymers are promising to mimic the topology of natural macrocycle-biomacromolecules due to their unique architecture of lacking chain ends. To systematically study the effect of glycopolymer architecture on the interactions with protein, the cyclic glycopolymers bearing galactose side-chain (cyclic PMAGn ) with three degrees of polymerization (n = 14, 24, 47) are prepared for the first time. The cyclic PMAGn exhibits unique properties in agglutinating and inhibiting proteins in subsequent studies by comparison with the linear precursor with the same molecular weights. More impressively, the cyclic PMAGn highlight the improved performance of cyclic architecture. For example, the cyclic PMAGn shows superior inhibition abilities to suppress amyloid formation from amyloid ß protein fragment 1-42 aggregation and block the specific interaction between bacteria and galactose-modified surface compared to that of respective linear counterpart. This interesting finding suggests that the architecture of cyclic glycopolymers may be capable of optimizing the ability to bind or inhibit proteins in biological processes.

Integration of CuAAC Polymerization and Controlled Radical Polymerization into Electron Transfer Mediated "Click-Radical" Concurrent Polymerization.

The successful chain-growth copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) polymerization employing Cu(0)/pentamethyldiethylenetriamine (PMDETA) and alkyl halide as catalyst is first investigated by a combination of nuclear magnetic resonance, gel-permeation chromatography, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. In addition, the electron transfer mediated "click-radical" concurrent polymerization utilizing Cu(0)/PMDETA as catalyst is successfully employed to generate well-defined copolymers, where controlled CuAAC polymerization of clickable ester monomer is progressed in the main chain acting as the polymer backbone, the controlled radical polymerization (CRP) of acrylic monomer is carried out in the side chain. Furthermore, it is found that there is strong collaborative effect and compatibility between CRP and CuAAC polymerization to improve the controllability.

One-Pot Multicomponent Synthesis of Glycopolymers through a Combination of Host-Guest Interaction, Thiol-ene, and Copper-Catalyzed Click Reaction in Water.

There is a common phenomenon that the heterogeneity of natural oligosaccharides contains various sugar units, which can be used to enhance affinity and selectivity toward a specific receptor, so the synthesis of heterogeneous glycopolymers is always an important issue in the glycopolymer field. Herein, this study conducts a one-pot method to prepare polyrotaxane-based heteroglycopolymers anchored with different sugar units and fluorescent moieties via the combination of host-guest interaction, thiol-ene, and copper-catalyzed click chemistry in water. Fourier transform infrared spectroscopy, nuclear magnetic resonance spectroscopy, gel permeation chromatography, X-ray diffraction, and Ellman's assay test are used in the paper to characterize the compounds. Quartz crystal microbalance-dissipation (QCD-D) experiments and bacterial adhesion assay are utilized to study the interactions of polyrotaxane-based heteroglycopolymers with Con A and Escherichia coli. The results reveal that polyrotaxanes (PRs) with mannose and glucose present better specificity toward Con A and E. coli than PRs with glucose due to synergistic effects.

Regenerable-Catalyst-Aided, Opened to Air and Sunlight-Driven "CuAAC&ATRP" Concurrent Reaction for Sequence-Controlled Copolymer.

An ideal stimuli-responsive controlled/living radical polymerization should have the ability to manipulate the reaction through spatiotemporal "on/off" controls, achieving the polymerization under fully open conditions and allowing for precise control over macromolecular architecture with defined molecular weights and monomer sequence. In this contribution, the photo (sunlight)-induced electron transfer atom transfer radical-polymerization (PET-ATRP) can be realized to be reversibly activated and deactivated under fully open conditions utilizing one-component copper(II) thioxanthone carboxylate as multifunctional photocatalyst and oxygen scavenger. The polymerization behaviors are investigated, presenting controlled features with first-order kinetics and linear relationships between molecular weights and monomer conversions. More importantly, "CuAAC&ATRP" concurrent reaction combining PET-ATRP, photodriven deoxygenation, and photoactivated CuAAC click reaction is successfully employed to synthesize the sequence-defined multiblock functional copolymers, in which the iterative monomer additions can be easily manipulated under fully open conditions.

Integrating Sugar and Dopamine into One Polymer: Controlled Synthesis and Robust Surface Modification.

Glycopolymers attached to a surface possess the ability to bind to certain carbohydrate binding proteins in a highly specific manner, and because of this, the fabrication of glycopolymer-modified surfaces has evolved as an effective route toward bioresponsive systems. Poly(N-3,4-dihydroxybenzenethyl methacrylamide-co-2-(methacrylamido) glucopyranose) copolymers, containing sugar and catechol functionalities, are for the first time successfully prepared in a well-controlled manner via room temperature single-electron transfer initiation and propagation through radical addition fragmentation chain transfer technique. The polymerization behavior is investigated and it presents controlled features with first-order kinetics and linear relationships between molecular weights and monomer conversions. Moreover, the copolymers are used to modify different types of surfaces (silicon, steel, and plastic), the properties of the surfaces and the specific lectin-binding abilities are investigated by a combination of water contact angle, Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectra, scanning electron microscopy with energy dispersive X-ray (SEM/EDX), atomic force microscopy, and confocal microscope measurements.

Ratiometric Fluorescent pH Probes Based on Glycopolymers.

Effectively detecting pH changes plays a critical role in exploring cellular functions and determining physiological and pathological processes. A novel ratiometric pH probe based on a glycopolymer, armored with properties of serum-stability, tumor-targeting, and pH monitoring, is designed. Random copolymers of 2-(methacrylamido) glucopyranose and fluorescein O-methacrylate are first synthesized by reversible addition fragmentation chain transfer polymerization. Acryloxyethyl thiocarbamoyl rhodamine B is then attached to the polymer chain to prepare ratiometric fluorescent pH probes via a thiol-ene reaction. The synthesized polymeric probes are characterized by NMR, gel permeation chromatography, UV-vis spectroscopy, and transmission electron microscopy, and the fluorescence responses are examined in phosphate buffer at different pHs. The cytotoxicity and confocal imaging experiments of the probes are detected using HeLa cells, demonstrating a low toxicity and superior biocompatibility for detecting pH changes in bioapplications.

Initiator and Photocatalyst-Free Visible Light Induced One-Pot Reaction: Concurrent RAFT Polymerization and CuAAC Click Reaction.

A new, visible light-catalyzed, one-pot and one-step reaction is successfully employed to design well-controlled side-chain functionalized polymers, by the combination of ambient temperature revisible addtion-fragmentation chain transfer (RAFT) polymerization and click chemistry. Polymerizations are well controlled in a living way under the irradiation of visible light-emitting diode (LED) light without photocatalyst and initiator, using the trithiocarbonate agent as iniferter (initiator-transfer agent-terminator) agent at ambient temperature. Fourier transfer infrared spectroscopy (FT-IR), NMR, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) data confirm the successful one-pot reaction. Compared to the reported zero-valent metal-catalyzed one-pot reaction, the polymerization rate is much faster than that of the click reaction, and the visible light-catalyzed one-pot reaction can be freely and easily regulated by turning on and off the light.

Regulation of Protein Binding Capability of Surfaces via Host-Guest Interactions: Effects of Localized and Average Ligand Density.

The protein binding capability of biomaterial surfaces can significantly affect subsequent biological responses, and appropriate ligand presentation is often required to guarantee the best functions. Herein, a new facile method for regulating this capability by varying the localized and average ligand density is presented. Binding between lysine and plasminogen relevant to a fibrinolysis system was chosen as a model. We integrated different lysine-modified ß-cyclodextrin (ß-CD) derivatives onto bioinert copolymer brushes via host-guest interactions. The localized and average lysine density can be conveniently modulated by changing the lysine valency on ß-CD scaffolds and by diluting lysine-persubstituted ß-CD with pure ß-CD, respectively. Both the plasminogen adsorption and the plasminogen binding affinity were enhanced by lysine-persubstituted ß-CD compared with those of lysine-monosubstituted ß-CD, which is possibly due to the higher localized lysine density and the multivalent binding of plasminogen on lysine-persubstituted ß-CD surfaces. With a change in the ratio of lysine-persubstituted ß-CD to ß-CD, the average lysine density can be tuned, leading to the linear regulation of the adsorption of plasminogen on surfaces.

Synthesis of Hemoglobin Conjugated Polymeric Micelle: A ZnPc Carrier with Oxygen Self-Compensating Ability for Photodynamic Therapy.

Photodynamic therapy (PDT) is a promising singlet oxygen ((1)O2) mediated clinical treatment for many tumors. As the source of (1)O2, oxygen plays an important role in the curative effect of PDT. However, the facts of photochemical depletion of oxygen and the intrinsic hypoxic microenvironment of tumors remain the major challenges. In this work, a novel photosensitizer carrier with oxygen self-compensating ability was designed for PDT. It was synthesized via chemical conjugation of hemoglobin (Hb) to polymeric micelles formed by triblock copolymers of poly(ethylene glycol)-block-poly(acrylic acid)-block-polystyrene (PEG-b-PAA-b-PS). The PEG-b-PAA-b-PS and resultant micelles in aqueous solution were comprehensively characterized by means of FTIR, (1)H NMR, GPC, DLS, TEM, and fluorescence spectroscopy. The oxygen-binding capacity and antioxidative activity of the Hb conjugated micelles were evaluated via UV-vis spectroscopy. In addition, compared with the control micelles without Hb, the Hb conjugated photosensitizer carrier was able to generate more (1)O2 and exert greater photocytotoxicity on Hela cells in vitro.