Gram-negative bacteria recognition and photodynamic elimination by Zn-DPA based sensitizers.

The abuse and overuse of antibiotics let drug-resistant bacteria emerges. Antibacterial photodynamic therapy (APDT) has shown outstanding merits to eliminate the drug-resistant bacteria via cytotoxic reactive oxygen species produced by irradiating photosensitizer. However, most of photosensitizers are not effective for Gram-negative bacteria elimination. Herein conjugates of NBS, a photosensitizer, linked with one (NBS-DPA-Zn) or two (NBS-2DPA-Zn) equivalents of zinc-dipicolylamine (Zn-DPA) have been designed to achieve the functional recognition of different bacteria. Due to the cationic character of NBS and metal transfer channel effect of Zn-DPA, NBS-DPA-Zn exhibited the first regent to distinguish P. aeruginosa from other Gram-negative bacteria. Whereas NBS-2DPA-Zn showed broad-spectrum antibacterial effect because the two arm of double Zn-DPA enhanced interactions with anionic membranes of bacteria, led the bacteria aggregation and thus provided the efficacy of APDT to bacteria and corresponding biofilm. In combination with a hydrogel of Pluronic, NBS-2DPA-Zn@gel shows promising clinical application in mixed bacterial diabetic mouse model infection. This might propose a new method that can realize functional identification and elimination of bacteria through intelligent regulation of Zn-DPA, and shows excellent potential for antibacterial application.

Dipicolylamine-Zn Induced Targeting and Photo-Eliminating of Pseudomonas aeruginosa and Drug-Resistance Gram-Positive Bacteria.

The emergence of drug-resistant bacteria, particularly resistant strains of Gram-negative bacteria, such as Pseudomonas aeruginosa, poses a significant threat to public health. Although antibacterial photodynamic therapy (APDT) is a promising strategy for combating drug-resistant bacteria, actively targeted photosensitizers (PSs) remain unknown. In this study, a PS based on dipicolylamine (DPA), known as WZK-DPA-Zn, is designed for the selective identification of P. aeruginosa and drug-resistant Gram-positive bacteria. WZK-DPA-Zn exploits the synergistic effects of DPA-Zn2+ coordination and cellular uptake, which could effectively anchor P. aeruginosa within a brief period (10 min) without interference from other Gram-negative bacteria. Simultaneously, the cationic nature of WZK-DPA-Zn enhances its interaction with Gram-positive bacteria via electrostatic forces. Compared to traditional clinical antibiotics, WZK-DPA-Zn shows exceptional antibacterial activity without inducing drug resistance. This effectiveness is achieved using the APDT strategy when irradiated with white light or sunlight. The combination of WZK-DPA-Zn with Pluronic-based thermosensitive hydrogel dressings (WZK-DPA-Zn@Gel) effectively eliminates mixed bacterial infections and accelerates wound healing, thereby achieving a synergistic effect where "1+1>2." In summary, this study proposes a precise strategy employing DPA-Zn as the targeting moiety of a PS, facilitating the rapid elimination of P. aeruginosa and drug-resistant Gram-positive bacteria using APDT.

Photocatalytic Antimicrobials: Principles, Design Strategies, and Applications.

Nowadays, the increasing emergence of antibiotic-resistant pathogenic microorganisms requires the search for alternative methods that do not cause drug resistance. Phototherapy strategies (PTs) based on the photoresponsive materials have become a new trend in the inactivation of pathogenic microorganisms due to their spatiotemporal controllability and negligible side effects. Among those phototherapy strategies, photocatalytic antimicrobial therapy (PCAT) has emerged as an effective and promising antimicrobial strategy in recent years. In the process of photocatalytic treatment, photocatalytic materials are excited by different wavelengths of lights to produce reactive oxygen species (ROS) or other toxic species for the killing of various pathogenic microbes, such as bacteria, viruses, fungi, parasites, and algae. Therefore, this review timely summarizes the latest progress in the PCAT field, with emphasis on the development of various photocatalytic antimicrobials (PCAMs), the underlying antimicrobial mechanisms, the design strategies, and the multiple practical antimicrobial applications in local infections therapy, personal protective equipment, water purification, antimicrobial coatings, wound dressings, food safety, antibacterial textiles, and air purification. Meanwhile, we also present the challenges and perspectives of widespread practical implementation of PCAT as antimicrobial therapeutics. We hope that as a result of this review, PCAT will flourish and become an effective weapon against pathogenic microorganisms and antibiotic resistance.

Construction of Rhodamine-Based AIE Photosensitizer Hydrogel with Clinical Potential for Selective Ablation of Drug-Resistant Gram-Positive Bacteria In Vivo.

The emergence of powerful antibiotic-resistant bacteria caused by the abuse of antibiotics has become a public health problem. Photodynamic antibacterial therapy is regarded as an innovative and promising antibacterial approach due to its minor side effects and lack of drug resistance. Nevertheless, few photosensitizers (PSs) are reported to have near-infrared (NIR) emission, the ability to rapidly discriminate bacteria, and high photodynamic antibacterial efficiency. In this study, it is reported for the first time that a water-soluble NIR fluorescence emission rhodamine-based photosensitizer with aggregation-inducing emission (AIE) effects, referred to as CS-2I, can efficiently identify and kill Gram-positive bacteria. In a fluorescence imaging experiment with blended bacteria, CS-2I can selectively target Gram-positive bacteria and specifically label Gram-positive bacteria with high efficiency after only 5 min of incubation. Furthermore, CS-2I achieves complete inhibition of methicillin-resistant Staphylococcus aureus (MRSA) at an extremely low concentration (0.5 µm) and light dosage (6 J cm-2 ). Remarkably, CS-2I is mixed with Carbomer 940 to prepare an antibacterial hydrogel dressing (CS-2I@gel), and in vitro and in vivo results demonstrate that CS-2I@gel provides extraordinary performance in photodynamic antibacterial therapy. Hence, this study provides a new strategy and blueprint for the future design of antibacterial materials.

Organic Photo-antimicrobials: Principles, Molecule Design, and Applications.

The emergence of multi-drug-resistant pathogens threatens the healthcare systems world-wide. Recent advances in phototherapy (PT) approaches mediated by photo-antimicrobials (PAMs) provide new opportunities for the current serious antibiotic resistance. During the PT treatment, reactive oxygen species or heat produced by PAMs would react with the cell membrane, consequently leaking cytoplasm components and effectively eradicating different pathogens like bacteria, fungi, viruses, and even parasites. This Perspective will concentrate on the development of different organic photo-antimicrobials (OPAMs) and their application as practical therapeutic agents into therapy for local infections, wound dressings, and removal of biofilms from medical devices. We also discuss how to design highly efficient OPAMs by modifying the chemical structure or conjugating with a targeting component. Moreover, this Perspective provides a discussion of the general challenges and direction for OPAMs and what further needs to be done. It is hoped that through this overview, OPAMs can prosper and will be more widely used for microbial infections in the future, especially at a time when the global COVID-19 epidemic is getting more serious.

A photo-sensitizable phage for multidrug-resistant Acinetobacter baumannii therapy and biofilm ablation.

Antibiotic abuse causes the emergence of bacterial resistance. Photodynamic antibacterial chemotherapy (PACT) has great potential to solve serious bacterial resistance, but it suffers from the inefficient generation of ROS and the lack of bacterial targeting ability. Herein, a unique cationic photosensitizer (NB) and bacteriophage (ABP)-based photodynamic antimicrobial agent (APNB) is developed for precise bacterial eradication and efficient biofilm ablation. Thanks to the structural modification of the NB photosensitizer with a sulfur atom, it displays excellent reactive oxygen species (ROS)-production ability. Moreover, specific binding to pathogenic microorganisms can be provided by bacteriophages. The developed APNB has multiple functions, including bacteria targeting, near-infrared fluorescence imaging and combination therapy (PACT and phage therapy). Both in vitro and in vivo experiments prove that APNB can efficiently treat A. baumannii infection. Particularly, the recovery from A. baumannii infection after APNB treatment is faster than that with ampicillin and polymyxin B in vivo. Furthermore, the strategy of combining bacteriophages and photosensitizers is employed to eradicate bacterial biofilms for the first time, and it shows the excellent biofilm ablation effect as expected. Thus, APNB has huge potential in fighting against multidrug-resistant bacteria and biofilm ablation in practice.

A long-wavelength ultrasensitive colorimetric fluorescent probe for carbon monoxide detection in living cells.

Exploring techniques for monitoring the intracellular signaling molecule carbon monoxide (CO) in biosystems is important to help understand its various cellular functions. Therefore, a simple long-wavelength colorimetric fluorescent probe LW-CO was designed for selectively and sensitively detecting intracellular CO in living systems. Probe LW-CO is ultrasensitive and can track CO levels in the range of 0-1 µM, with a detection limit of about 3.2 nM. Additionally, the obvious color changes of probe LW-CO with CO (yellow to pink) provide a convenient way for on-site detection of CO with the naked eye. Probe LW-CO was applied to track the exogenous levels of CO in RAW264.7 cells. Probe LW-CO proved to be an efficient method for investigating various cellular functions of CO.

A highly specific and ultrasensitive p-aminophenylether-based fluorescent probe for imaging native HOCl in live cells and zebrafish.

It is very important to detect native hypochlorous acid (HOCl) in the complex biosystems owing to the important roles of HOCl in the immune defense and the pathogenesis of numerous diseases. In this paper, a new p-aminophenylether-based fluorescent probe PAPE-HA was developed for specific detection of HOCl. Probe PAPE-HA could implement the quantitative detection of HOCl ranging from 0 to 1 µM and the detection limit was obtained as low as 1.37 nM. Additionally, probe PAPE-HA could reach a rapid response for HOCl (<2 min). Importantly, probe PAPE-HA with preeminent specificity and ultrasensitivity was proven to possess powerful capability of tracking native HOCl in live cells and zebrafish, and we thus anticipate that probe PAPE-HA could be used as a novel promising tool for revealing diverse cellular functions of HOCl.

A carbonothioate-based far-red fluorescent probe for the specific detection of mercury ions in living cells and zebrafish.

The detection of ionic mercury (Hg2+) is very important because it is a highly toxic environmental pollutant that could cause serious diseases and threaten human health. Herein, we designed a new carbonothioate-based far-red fluorescent probe, CBRB, with a seminaphthorhodafluor dye as the fluorophore for the detection of Hg2+. The CBRB probe by itself exhibited very weak fluorescence due to the enhanced photo-induced electron transfer (PET) effect and inhibited the intramolecular charge transfer (ICT) process caused by the carbonothioate moiety. Upon addition of Hg2+, a tremendous fluorescence enhancement was achieved, attributed to the removal of the carbonothioate group via a specific mercury-promoted desulfurization reaction. Moreover, the probe displayed a large Stokes shift (about 105 nm) and was used to quantitatively measure the concentration of Hg2+ for concentrations ranging from 0 to 1 µM (DL = 3.6 nM). In addition, CBRB in our experiments responded exclusively to Hg2+, even in the presence of high concentrations other ions. Gratifyingly, this probe was successfully used to monitor Hg2+ in environmental water samples and to image Hg2+ in living cells as well as in zebrafish.

A highly selective and ultrasensitive ratiometric fluorescent probe for peroxynitrite and its two-photon bioimaging applications.

In this work, taking full advantage of the intramolecular charge transfer (ICT) mechanism, a hydroxynaphthalimide-based ratiometric two-photon fluorescent probe RTP-PN was synthesized to detect ONOO-. Probe RTP-PN could accurately detect ONOO- in the range of 1.4 nM-1.4 µM with the detection limit of 1.4 nM by a ratiometric fluorescence spectroscopy method. Additionally, probe RTP-PN exhibited an ultrafast response for ONOO- than other various species including H2O2 and ClO-. Finally, probe RTP-PN was successfully adopted to detect intracellular ONOO- by the two-photon excitation microscopy.

Rational Design of a Hepatoma-Specific Fluorescent Probe for HOCl and Its Bioimaging Applications in Living HepG2 Cells.

Liver cancer is a kind of high mortality cancer due to the difficulty of early diagnosis. And according to the reports, the concentration of reactive oxygen species (ROS) was higher in cancer cells than normal cells. Therefore, developing an effective fluorescent probe for hepatoma-selective imaging of hypochlorous acid (HOCl) which is one of the vital ROS is of great importance for understanding the role of HOCl in liver cancer pathogenesis. However, the cell-selective fluorescent probe still remains a difficult task among current reports. Herein, a galactose-appended naphthalimide (Gal-NPA) with p-aminophenylether as a new receptor and galactose moiety as hepatoma targeting unit was synthesized and employed to detect endogenous HOCl in living HepG2 cells. This probe was proved to possess good water solubility and could respond specifically to HOCl. In addition, probe Gal-NPA could completely react to HOCl within 3 s meanwhile accompanied by tremendous fluorescence enhancement. The quantitative linear range between fluorescence intensities and the HOCl concentrations was 0 to 1 µM (detection limit = 0.46 nM). More importantly, fluorescence confocal imaging experiments showed that probe Gal-NPA could discriminate hepatoma cells over other cancer cells and simultaneously trace endogenous HOCl levels in living HepG2 cells. And we thus anticipate that probe Gal-NPA has the potential application for revealing the functions of HOCl in hepatoma cells.

A highly selective colorimetric and long-wavelength fluorescent probe for the detection of Hg2.

Currently, the fluorescent probe is an important method for detecting heavy metal ions, especially mercury ion (Hg2+ ), which is harmful to the health of humans and the environment due to its toxicity and extensive use. In this paper, we designed and synthesized a colorimetric and long-wavelength fluorescent probe Hg-P with high sensitivity and excellent selectivity, which could detect Hg2+ by the changes of visual color, fluorescence and absorption spectroscopy. With the addition of Hg2+ to probe Hg-P solution, its color changed from yellow to pink, and showed a 171 nm red-shifted absorption spectrum. Probe Hg-P was used in real water and soil solution samples to detect Hg2+ , and the result is satisfactory. Therefore, this new probe shows great value and application in detecting Hg2+ in the environment.

A Colorimetric and Fluorescent Probe for the Detection of Cu2+ in a Complete Aqueous Solution.

The fluorescent probe has become an important method for the detection of heavy metal ions. In the present work, a new and simple fluorescent probe, Cu-P, for detecting copper ion (Cu2+) was designed and synthesized. The probe has shown high sensitivity and selectivity toward Cu2+. The detection limit was 13 nM (based on the 3σ/slope). A significant color change from yellow to pink was observed; thus, the probe Cu-P could serve as a "naked-eye" indicator for Cu2+. Furthermore, the proposed probe was used to detect Cu2+ in real water and soil extract samples, with the result being satisfactory. Therefore, our proposed probe would provide a promising method for the detection of Cu2+ in the environment.

A highly specific and ultrasensitive near-infrared fluorescent probe for imaging basal hypochlorite in the mitochondria of living cells.

The development of highly specific and ultrasensitive fluorescent probes for tracking basal mitochondrial hypochlorite is very important to unravel its diverse cellular functions in the mitochondria of living cells. In this paper, we have developed a water-soluble, mitochondria-targeted near-infrared fluorescent probe NB-OCl for selectively measuring OCl- in the presence of higher concentration (500 µM) other biologically important substances. Surprisingly, the obtained results demonstrated that probe NB-OCl could sensitively determine OCl- in the range of 0-200 pM with the detection limit of 10.8 pM. To the best of our knowledge, NB-OCl is the first fluorescent probe for the specific determination of OCl- at the picomolar level. Moreover, probe NB-OCl exhibits a fast response for OCl- (< 5 s), which would be in favor of tracking the highly reactive and short-lived OCl- in the living systems. The preeminent recognition properties of probe NB-OCl enable its applications in the monitoring of basal OCl- and the fluctuations of endogenous/exogenous OCl- levels in the mitochondria of living cells.

A carbonothioate-based highly selective fluorescent probe with a large Stokes shift for detection of Hg2.

Mercury (Hg) is one of the heavy metal pollutants in the environment. Even a very small amount of mercury can cause serious harm to human beings. Herein, we reported a new carbonothioate-based fluorescent probe for the detection of Hg2+ without interference from other metal ions. This probe possessed a very large Stokes shift (192 nm), which could improve the detection sensitivity by minimizing the interferences resulted from self-absorption or auto-fluorescence. With the addition of Hg2+ to the probe solution, considerable fluorescence enhancement was observed. Additionally, the Hg2+ concentration of 0-16 µM and fluorescence intensity showed a good linear relationship (y = 22106× + 53108, R2  = 0.9955). Finally, the proposed probe was used to detect Hg2+ in real water samples, and its result was satisfactory. Therefore, our proposed probe would provide a promising method for the determination of Hg2+ in the environment.

A highly specific and ultrasensitive fluorescent probe for basal lysosomal HOCl detection based on chlorination induced by chlorinium ions (Cl+).

The development of techniques for detecting HOCl at the subcellular level is very important to elucidate its cellular functions. Due to its relatively low concentration, it is still a great challenge to specifically track the basal HOCl in normal cells. In this paper, based on the unique chlorination of HOCl by the initiation of chlorinium ions (Cl+) in an acidic medium, we have developed a simple pH-mediated lysosome-targetable fluorescent probe Lyso-HOCl for the specific detection of HOCl over other bioactive molecules at higher concentration (500 µM). Our results show that Lyso-HOCl possesses a detection limit of 8.0 pM, and can quantitatively detect HOCl at the picomolar level. The ultrasensitive and ultrafast response property of probe Lyso-HOCl offers a good opportunity to monitor the basal HOCl and the fluctuation of endogenous HOCl levels in the lysosomes of macrophages (Raw 264.7 cells), and we thus anticipate that this probe would provide a promising tool for further unraveling the biological functions of HOCl in subcellular lysosomes.

Highly Specific and Ultrasensitive Two-Photon Fluorescence Imaging of Native HOCl in Lysosomes and Tissues Based on Thiocarbamate Derivatives.

Hypochlorous acid (HOCl) acts as a weak acid distributed mainly in acidic organelle lysosomes of phagocytes and plays a crucial role in the immune defense. The elaborate interrelation between the variations of HOCl levels in lysosomes and different physiological and pathological processes remains unclear. Thus, the accurate determination of lysosomal HOCl in living cells and in vivo is very important. Because of extremely low concentration and difficulty in distinguishing HOCl from OCl- under the physiological environment, it is still a great challenge to specifically monitor the intracellular intrinsic HOCl levels without exogenous stimulation, which impedes an exact understanding of its biological roles. In this paper, based on the electrophilic addition of Cl+ to sulfide moiety, we have developed a two-photon fluorescent probe O-(N-butyl-1,8-naphthalimide)-4-yl-N,N-dimethylthiocarbamate (NDMTC) for the specific determination of HOCl over OCl- and other bioactive molecules. Our results show that NDMTC possesses a detection limit of 7.6 pM, and it is the first fluorescent probe for detecting HOCl at the picomolar level. Furthermore, by introducing an alkylmorpholine group to the NDMTC framework, the lysosome-targetable derivative Lyso-NDMTC was obtained, and its ability to image HOCl in the lysosome organelles was clearly confirmed. Combined with two-photon fluorescence imaging of background suppression and deeper tissue penetration, NDMTC and Lyso-NDMTC were used to successfully visualize intracellular native HOCl and discern tumor tissue in mice. This study offers two perfect fluorescence imaging probes for further investigation of pathological roles of HOCl in various diseases.

A fast-response, highly sensitive and selective fluorescent probe for the ratiometric imaging of nitroxyl in living cells.

A fast-response, highly sensitive and selective fluorescent probe with the 2-(diphenylphosphino)benzoate moiety as a recognition receptor for the ratiometric imaging of nitroxyl in living cells was first developed.