pH-Responsive Fluorescent Polymer–Drug System for Real-Time Detection and <i>In Situ</i> Eradication of Bacterial Biofilms

Bacterial biofilms encased in extracellular polymeric substances to create protected microenvironments are typically challenging to disperse by common antibiotics and cannot be in situ visualized under current modalities. Herein, a pH-responsive branched polymer [poly­(MBA-AEPZ)-AEPZ-NA] capable of overcoming antibiotic resistance and real-time visualizing biofilms for fluorescence imaging-guided infection control is reported. The positively charged polymer can effectively penetrate bacterial biofilms, neutralize the anionic character, and then disrupt the structural integrity, thus significantly promoting the transport of antibiotics into biofilms. The polymer shows a weak fluorescence emission intensity under physiological conditions (pH 7.4) but emits intense green-light emission within the localized biofilm microenvironment (pH 5.5) to real-time visualize bacterial biofilms. A therapeutic system made of the polymer and a model antibiotic can significantly reduce the dosages of the drug, thereby minimizing biofilm-induced drug resistance. Notably, a green fluorescent polymer responding to localized pH conditions is demonstrated in living zebrafish. This work confirmed that combinations of the pH-responsive branched polymer and antibiotics could be administered to overcome drug resistance and realize fluorescence imaging-guided treatment of bacterial biofilm infections

Dual-Emission Carbonized Polymer Dots for Ratiometric pH Sensing, pH-Dependent Generation of Singlet Oxygen, and Imaging-Guided Dynamics Monitoring of Photodynamic Therapy

The pH environment in cancer cells has been demonstrated to display vital influences on the therapeutic effect of photodynamic therapy (PDT). It is very interesting to develop pH-responsive probes for simultaneous pH sensing and dynamics monitoring of the effects of PDT, and therefore assessing the correlation between them. In this study, a multifunctional fluorescence probe, dual-emission carbonized polymer dot (CPD) in blue and red regions, which uses ethylene imine polymer (PEI) and 4,4′,4″,4‴-(porphine-5, 10, 15, 20-tetrayl) tetrakis (benzoic acid) (TCPP) as precursors through a one-step hydrothermal amide reaction, has been designed for ratiometric pH sensing, generating pH-dependent 1O2 for PDT of cancer cells, and investigating the dynamics effects of PDT through pH-guided imaging. The prepared CPDs were successfully used for ratiometric pH response, pH-dependent generation of 1O2, and dynamics monitoring PDT in HeLa cells. This study may provide an alternative strategy to prepare CPD-based theranostic integrated nanoprobes for PDT through the rational design of precursors

pH-Responsive Hyperbranched Polymer Nanoparticles to Combat Intracellular Infection by Disrupting Bacterial Wall and Regulating Macrophage Polarization

Intracellular bacterial infections pose a serious threat to public health. Macrophages are a heterogeneous population of immune cells that play a vital role in intracellular bacterial infection. However, bacteria that survive inside macrophages could subvert the cell signaling and eventually reduce the antimicrobial activity of macrophages. Herein, dual pH-responsive polymer (poly[(3-phenylprop-2-ene-1,1-diyl)bis(oxy)bis(enthane-2,1-diyl)diacrylate-co-N-aminoethylpiperazine] (PCA)) nanoparticles were developed to clear intracellular bacteria by activating macrophages and destructing bacterial walls. The presence of acid-labile acetal linkages and tertiary amine groups in the polymer’s backbone endow hyperbranched PCA dual pH-response activity that shows acid-induced positive charge increase and cinnamaldehyde release properties. The biodegraded PCA nanoparticles could significantly inhibit the growth of bacteria by damaging the bacterial walls. Meanwhile, PCA nanoparticles could uptake by macrophages, generate reactive oxygen species (ROS), and remodel the immune response by upregulating M1 polarization, leading to the reinforced antimicrobial capacity. Furthermore, PCA nanoparticles could promote bacteria-infected wound healing in vivo. Therefore, these dual pH-responsive PCA nanoparticles enabling bacteria-killing and macrophage activation provide a novel outlook for treating intracellular infection

General Strategy to Achieve Color-Tunable Ratiometric Two-Photon Integrated Single Semiconducting Polymer Dot for Imaging Hypochlorous Acid

It is highly desired and challenging to construct integrated (all-in-one) single semiconducting-polymer-derived dot (Pdot) without any postmodification but with desired performances for bioapplications. In this work, eight hypochlorous acid (HClO)-sensitive integrated polymers and corresponding polymer-derived Pdots are designed through molecular engineering to comparatively study their analytical performances for detecting and imaging HClO. The optimized polymers-derived Pdots are obtained through regulating donor–acceptor structure, the content of HClO-sensitive units, and the position of HClO-sensitive units in the polymer backbone. The designed Pdots display distinguished characteristics including multicolours with blue, yellow, and red three primary fluorescence colors, determination mode from single-channel to dual-channel (ratiometric) quantification, ultrafast response, low detection limit, and high selectivity for ClO– sensing based on specific oxidation of ClO–-sensitive unit 10-methylphenothiazine (PT) accompanied by altering the intramolecular charge transfer (ICT) and fluorescence resonance energy transfer (FRET) processes in Pdots. The prepared integrated Pdots are also applied for two-photon ClO– imaging in HeLa cells and one- and two-photon ClO– imaging produced in acute inflammation in mice with satisfactory results. We believe that the present study not only provides excellent integrated fluorescent nanoprobes for ClO– monitoring in living systems but also extends a general strategy for designing integrated semiconducting polymers and Pdots with desired performances for biological applications

ε‑Polylysine-Based Macromolecules with Catalase-Like Activity to Accelerate Wound Healing by Clearing Bacteria and Attenuating Inflammatory Response

Wound healing has remained a critical challenge due to its susceptibility to bacterial infection and the unique biological inflammatory response. Safe and effective therapeutics are still lacking. Biodegradable macromolecules (ε-polylysine-g-ferrocene, EPL-g-Fc) were developed to accelerate wound healing by combating bacterial infection and attenuating inflammatory responses. The biodegradable macromolecules were prepared via a Schiff-based reaction between ferrocene carboxaldehyde (Fc) and ε-polylysine (EPL). Through the synergistic combination of positive-charged EPL and π–π stacked Fc, the macromolecules possess excellent antibacterial activities. EPL-g-Fc with catalase-like activity could modulate the oxidative microenvironment in mammalian cells and zebrafish by catalyzing H2O2 into H2O and O2. EPL-g-Fc could alleviate inflammatory response in vitro. Furthermore, the macromolecules could accelerate bacteria-infected wound healing in vivo. This work provides a versatile strategy for repairing bacteria-infected wounds by eliminating bacteria, modulating oxidative microenvironment, and alleviating inflammatory response

Cascade-Targeted Nanoplatforms for Synergetic Antibiotic/ROS/NO/Immunotherapy against Intracellular Bacterial Infection

Intracellular bacteria in dormant states can escape the immune response and tolerate high-dose antibiotic treatment, leading to severe infections. To overcome this challenge, cascade-targeted nanoplatforms that can target macrophages and intracellular bacteria, exhibiting synergetic antibiotic/reactive oxygen species (ROS)/nitric oxide (NO)/immunotherapy, were developed. These nanoplatforms were fabricated by encapsulating trehalose (Tr) and vancomycin (Van) into phosphatidylserine (PS)-coated poly[(4-allylcarbamoylphenylboric acid)-ran-(arginine-methacrylamide)-ran-(N,N′-bisacryloylcystamine)] nanoparticles (PABS), denoted as PTVP. PS on PTVP simulates a signal of “eat me” to macrophages to promote cell uptake (the first-step targeting). After the uptake, the nanoplatform in the acidic phagolysosomes could release Tr, and the exposed phenylboronic acid on the nanoplatform could target bacteria (the second-step targeting). Nanoplatforms can release Van in response to infected intracellular overexpressed glutathione (GSH) and weak acid microenvironment. l-arginine (Arg) on the nanoplatforms could be catalyzed by upregulated inducible nitric oxide synthase (iNOS) in the infected macrophages to generate nitric oxide (NO). N,N′-Bisacryloylcystamine (BAC) on nanoplatforms could deplete GSH, allow the generation of ROS in macrophages, and then upregulate proinflammatory activity, leading to the reinforced antibacterial capacity. This nanoplatform possesses macrophage and bacteria-targeting antibiotic delivery, intracellular ROS, and NO generation, and pro-inflammatory activities (immunotherapy) provides a new strategy for eradicating intracellular bacterial infections

Near-Infrared Light-Activated Thermosensitive Liposomes as Efficient Agents for Photothermal and Antibiotic Synergistic Therapy of Bacterial Biofilm

Biofilm is closely related to chronic infections and is difficult to eradicate. Development of effective therapy strategies to control biofilm infection is still challenging. Aiming at biofilm architecture, we designed and prepared near-infrared-activated thermosensitive liposomes with photothermal and antibiotic synergistic therapy capacity to eliminate Pseudomonas aeruginosa biofilm. The liposomes with positive charge and small size aided to enter the biofilm microchannels and locally released antibiotics in infection site. The liposomes could remain stable at 37 °C and release about 80% antibiotics over 45 °C. The biofilm dispersion rate was up to 80%, which was a 7- to 8-fold rise compared to excess antibiotic alone, indicating that the localized antibiotic release and photothermal co-therapy improved the antimicrobial efficiency. In vivo drug-loaded liposomes in treating P. aeruginosa-induced abscess exhibited an outstanding therapeutic effect. Furthermore, photothermal treatment could stimulate the expression of bcl2-associated athanogene 3 to prevent normal tissue from thermal damage. The near-infrared-activated nanoparticle carriers had the tremendous therapeutic potential to dramatically enhance the efficacy of antibiotics through thermos-triggered drug release and photothermal therapy

Bioactive Peptide-Mimicking Polymer Nanoparticles for Bacteria Imaging and Chemo/Immunotherapy of Intracellular Infection

Intracellular bacterial infections are extremely difficult to be treated because intracellular bacteria have developed resistant mechanisms to escape the immune attack and antibiotic therapy. It remains challenging to develop antibiotic-free materials and relative strategies for treating intracellular bacterial infections. Herein, a new host-defense peptide-mimicking polymer nanoparticle, inspired by cell-penetrating peptides, was developed to eradicate intracellular bacteria by its outstanding antibacterial and pro-inflammatory immunomodulatory. The polymer nanoparticle (TPE-Parg) was prepared through ring-opening polymerization of N6-carbobenzoxy-l-lysine N-carboxyanhydride (Cbz-l-Lys NCA) using 1-(4-aminophenyl)-1,2,2-triphenylethene (TPE-NH2) as the initiator, followed by deprotection of the Cbz-l-Lys NCA group and guanidinium modification. The impact of cationic functional groups and chain length variation on the antibacterial activity of polymer nanoparticle were investigated in detail. The results confirmed that the optimal polymer nanoparticle could not only image bacteria with aggregation-induced blue fluorescence, but also kill planktonic bacteria with low cytotoxicity. Furthermore, the nanoparticle could induce macrophages to generate nitric oxide (NO) and activate the immune system to eliminate intracellular bacteria. The nanoparticle further showed its potent in vivo antibacterial activity in an intracellular Staphylococcus aureus infection model fabricated on mice hypodermic. The obtained multifunctional host-defense peptide-mimicking polymer nanoparticles with potent antibacterial activity (chemotherapy) and pro-inflammatory immunomodulatory (immunotherapy) are excellent alternatives for intracellular antibacterial therapy and provide a direction for developing innovative antimicrobials