Activatable probes with potential for intraoperative tumor-specific fluorescence-imaging guided surgery.

Owing to societal development and aging population, the impact of cancer on human health and quality of life has increased. Early detection and surgical treatment are the most effective approaches for most cancer patients. As the scope of conventional tumor resection is determined by auxiliary examination and surgeon experience, there is often insufficient recognition of tiny tumors. The ability to detect such tumors can be improved by using fluorescent tumor-specific probes for surgical navigation. This review mainly describes the design principles and mechanisms of activatable probes for the fluorescence imaging of tumors. This type of probe is nonfluorescent in normal tissue but exhibits obvious fluorescence emission upon encountering tumor-specific substrates, such as enzymes or bioactive molecules, or changes in the microenvironment, such as a low pH. In some cases, a single-factor response does not guarantee the effective fluorescence labeling of tumors. Therefore, two-factor-activatable fluorescence imaging probes that react with two specific factors in tumor cells have also been developed. Compared with single biomarker testing, the simultaneous monitoring of multiple biomarkers may provide additional insight into the role of these substances in cancer development and aid in improving the accuracy of early cancer diagnosis. Research and progress in this field can provide new methods for precision medicine and targeted therapy. The development of new approaches for early diagnosis and treatment can effectively improve the prognosis of cancer patients and help enhance their quality of life.

A self-assembled nanophotosensitizer targets lysosomes and induces lysosomal membrane permeabilization to enhance photodynamic therapy.

We report the self-assembly of amphiphilic BDQ photosensitizers into lysosome-targeting nanophotosensitizer BDQ-NP for highly effective photodynamic therapy (PDT). Molecular dynamics simulation, live cell imaging, and subcellular colocalization studies showed that BDQ strongly incorporated into lysosome lipid bilayers to cause continuous lysosomal membrane permeabilization. Upon light irradiation, the BDQ-NP generated a high level of reactive oxygen species to disrupt lysosomal and mitochondrial functions, leading to exceptionally high cytotoxicity. The intravenously injected BDQ-NP accumulated in tumours to achieve excellent PDT efficacy on subcutaneous colorectal and orthotopic breast tumor models without causing systemic toxicity. BDQ-NP-mediated PDT also prevented metastasis of breast tumors to the lungs. This work shows that self-assembled nanoparticles from amphiphilic and organelle-specific photosensitizers provide an excellent strategy to enhance PDT.

AIPH-Encapsulated Thermo-Sensitive Liposomes for Synergistic Microwave Ablation and Oxygen-Independent Dynamic Therapy.

Microwave ablation (MWA) is a novel treatment modality that can lead to the death of tumor cells by heating the ions and polar molecules in the tissue through high-speed vibration and friction. However, the single hyperthermia is not sufficient to completely inhibit tumor growth. Herein, a thermodynamic cancer-therapeutic modality has been fabricated which could be able to overcome hypoxia's limitations in the tumor microenvironment. Using thermo-sensitive liposomes (TSLs) and oxygen-independent radical generators (2,2'-azobis[2-(2-imidazolin-2-yl)propane]dihydrochloride [AIPH]), a nano-drug delivery system denoted as ATSL is developed for efficient sequential cancer treatment. Under the microwave field, the temperature rise of local tissue could not only lead to the damage of tumor cells but also induce the release of AIPH encapsulated in ATSL to produce free radicals, eliciting tumor cell death. In addition, the ATSL developed here would avoid the side effects caused by the uncontrolled diffusion of AIPH to normal tissues. The ATSLs have shown excellent therapeutic effects both in vitro and in vivo, suggesting its highly promising potential for clinic.

Pseudocatalytic Hydrogels with Intrinsic Antibacterial and Photothermal Activities for Local Treatment of Subcutaneous Abscesses and Breast Tumors.

The intimate relationship between bacteria and tumors has triggered a lot of activities in the development and design of bioactive materials to concurrently respond to antitumor and antibacterial demands. Herein, a pseudocatalytic hydrogel (AM-I@Agar) with intrinsic antibacterial and photothermal activities, synthesized by incorporating prefabricated amylose-iodine nanoparticles into low-melting-point agarose hydrogel, is explored as a bioactive agent for local treatment of subcutaneous abscesses and breast tumors. The AM-I@Agar hydrogel depicts the ability of pseudocatalytic O2 generation from H2 O2 to alleviate hypoxia. Meanwhile, the AM-I@Agar hydrogel exhibits temperature self-regulation features, beneficial for avoiding thermal injury during photothermal therapy owing to thermochromic properties. Upon local injection into a subcutaneous abscess, methicillin-resistant Staphylococcus aureus is effectively eliminated by the AM-I@Agar hydrogel, and complete skin recovery is achieved in 8 d, demonstrating much better antibacterial effects compared with penicillin, a small-molecule antibiotic. AM-I/5-FU@Agar hydrogel, obtained after loading 5-fluorouracil (5-FU), significantly inhibits tumors in both normal 4T1 tumor-bearing mice and MRSA-infected 4T1 tumor-bearing mice models via a synergistic photothermal-chemo effect, and shows treatment efficiency superior to that achieved with photothermal therapy or 5-FU alone. This work provides a concept for the design and development of bioactive agents for potential management of bacteria-associated cancer.

Ultrathin Hafnium Disulfide Atomic Crystals with ROS-Scavenging and Colon-Targeting Capabilities for Inflammatory Bowel Disease Treatment.

The exciting success of NBTXR3 in the clinic has triggered a tumult of activities in the design and development of hafnium-based nanoparticles. However, due to the concerns of nondegradation and limited functions, the biomedical applications of Hf-based nanoparticles mainly focus on tumors. Herein, tannic acid capped hafnium disulfide (HfS2@TA) nanosheets, a 2D atomic crystal of hafnium-based materials prepared by liquid-phase exfoliation, were explored as high-performance anti-inflammatory nanoagents for the targeted therapy of inflammatory bowel disease (IBD). Benefiting from the transformation of the S2-/S6+ valence state and huge specific surface area, the obtained HfS2@TA nanosheets were not only capable of effectively eliminating reactive oxygen species/reactive nitrogen species and downregulating pro-inflammatory factors but also could be excreted via kidney and hepatointestinal systems. Unexpectedly, HfS2@TA maintained excellent targeting capability to an inflamed colon even in the harsh digestive tract environment, mainly attributed to the electrostatic interactions between negatively charged tannic acid and positively charged inflamed epithelium. Encouragingly, upon oral or intravenous administration, HfS2@TA quickly inhibited inflammation and repaired the intestinal mucosa barrier in both dextran sodium sulfate and 2,4,6-trinitrobenzenesulfonic acid induced IBD models. This work demonstrated that ultrathin HfS2@TA atomic crystals with enhanced colon accumulation were promising for the targeted therapy of IBD.

Nanoscale Metal-Organic Layers Detect Mitochondrial Dysregulation and Chemoresistance via Ratiometric Sensing of Glutathione and pH.

Mitochondrial dysregulation controls cell death and survival by changing endogenous molecule concentrations and ion flows across the membrane. Here, we report the design of a triply emissive nanoscale metal-organic layer (nMOL), NA@Zr-BTB/F/R, for sensing mitochondrial dysregulation. Zr-BTB nMOL containing Zr6 secondary building units (SBUs) and 2,4,6-tris(4-carboxyphenyl)aniline (BTB-NH2) ligands was postsynthetically functionalized to afford NA@Zr-BTB/F/R by exchanging formate capping groups on the SBUs with glutathione(GSH)-selective (2E)-1-(2'-naphthyl)-3-(4-carboxyphenyl)-2-propen-1-one (NA) and covalent conjugation of pH-sensitive fluorescein (F) and GSH/pH-independent rhodamine-B (R) to the BTB-NH2 ligands. Cell imaging demonstrated NA@Zr-BTB/F/R as a ratiometric sensor for mitochondrial dysregulation and chemotherapy resistance via GSH and pH sensing.

BODIPY-based asymmetric monosubstituted (turn-on) and symmetric disubstituted (ratiometric) fluorescent probes for selective detection of phosgene in solution and gas phase.

Here, we designed and synthesized two fluorescent probes for detecting phosgene by nucleophilic substitution reaction using BODIPY as a fluorophore and 2-aminobenzylamine as reactive functional group. For the first time, we have studied the similarities and differences between asymmetric monosubstituted (1) and symmetric disubstituted (2) probes. A monosubstituted probe 1 (having a 2-aminobenzylamine group at the 3-position of BODIPY) has fluorescence-enhancing (weak green fluorescence to strong green fluorescence) responce to phosgene in 30 s with a large Stokes shift (∼70 nm) and sensitive property (DL = 0.81 nM); while the disubstituted probe 2 (having two 2-aminobenzylamine groups at the 3, 5-position of BODIPY) has a ratiometric fluorescent responce to phosgene in 2 min. The linear range of the response is wider than that of the monosubstitution probe 1, and the detection limit is also lower (2.36 nM). In addition, probes 1 and 2 can effectively eliminate the interference of other substances with similar chemical structure as phosgene. They can not only detect phosgene in solution environment, but also in gaseous environment quickly and sensitively.

A naphthalimide-based lysosome-targeting fluorescent probe for the selective detection and imaging of endogenous peroxynitrite in living cells.

The morpholine (ML) group can be used as a targeting unit for lysosomes. Here, a novel turn-off fluorescence probe for the highly selective imaging of peroxynitrite (ONOO-) produced by the endogenous stimulation of lysosomes in living cells is presented. The probe, denoted ML-NAP-DPPEA, comprises ML and 2-(diphenylphosphino)ethylamine (DPPE) groups attached to the fluorophore naphthalimide (NAP). ML-NAP-DPPEA shows excellent properties, including high selectivity for ONOO-, low cytotoxicity, and no interference, leading to low detection limits (17.6 nM). In the presence of ONOO-, the secondary amine group (NH) is oxidized to an electron-withdrawing group (HN → O), which quenches the fluorescence of ML-NAP-DPPEA. This intracellular lysosomal imaging technique was tested, and the results pointed to its potential use as a probe for studying the biological function and pathological effects of ONOO- in subcellular structure. Graphical abstract.

A ratiometric fluorescent probe for detection of endogenous and exogenous hydrogen sulfide in living cells.

A ratiometric visualized fluorescent probe of H2S of intramolecular charge transfer (ICT) and excited intramolecular proton transfer (ESIPT) mechanisms due to solvation effects has been designed and synthesized. This chemosensor shows the distinct signal changes with dual-emission in blue and green fluorescence spectral channel (from 495 nm to 525 nm) upon the addition of H2S in a single wavelength excitation. This chemosensor exhibits the low detection limit (91 nM) and high sensitivity and selectivity. Based on this, this chemosensor was successfully used not only to monitor H2S exogenously but also used to detect and image the endogenously generated H2S in HeLa cells with excellent performance.

Mechanochromic luminescent covalent organic frameworks for highly selective hydroxyl radical detection.

The mechanochromic luminescence phenomenon was first reported in the low-cost mechanochemical (MC) synthesis process of a covalent organic framework (COF-TpMA (MC)) prepared using triformylphloroglucinol (Tp) and melamine (MA) as precursors, which shows excellent performance as a hydroxyl radical (˙OH) detector in living systems. This work not only shows a new kind of mechanochromic luminescent material but also exhibits a close relationship between the π-π stacking and luminescence properties of COF materials.

BODIPY-based fluorescent sensor for imaging of endogenous formaldehyde in living cells.

A new highly selective fluorescent chemosensor for formaldehyde (FA) has been synthesized based on boron dipyrromethene as fluorophore and o-phenylenediamine (OPDA) as reaction group. When FA is added, the fluorescence emission band of the chemosensor red shift (from 525 nm to 548 nm) accompanied by an increase in intensity with strong green fluorescence was observed. This chemosensor also exhibited the lowest detection limit (0.104 µM) distinguished with other articles that have been reported. The application to cellular fluorescence imaging or test papers detection both indicated that the probe was highly responsive to the FA in endogenous cells and in the gaseous environment.

Fast and Selective Two-Stage Ratiometric Fluorescent Probes for Imaging of Glutathione in Living Cells.

Two fluorescent, m-nitrophenol-substituted difluoroboron dipyrromethene dyes have been designed by nucleophilic substitution reaction of 3,5-dichloro-4,4-difluoro-4-bora-3a,4a-diaza-s-indacene (BODIPY). Nonsymmetric and symmetric probes, that is. BODIPY 1 (with one nitrophenol group at the position 3) and BODIPY 2 (with two nitrophenol groups at the positions 3 and 5) were applied to ratiometric fluorescent glutathione detection. The detection is based on the two-step nucleophilic aromatic substitution of the nitrophenol groups of the probes by glutathione in buffer solution containing CTAB. In the first stage, probe 1 showed ratiometric fluorescent color change from green (λem = 530 nm) to yellow (λem = 561 nm) because of monosubstitution with glutathione (I561nm/I530nm). Addition of excess glutathione caused the second stage of ratiometric fluorescent color change from yellow to reddish orange (λem = 596 nm, I596nm/I561nm) due to disubstitution with glutathione. Therefore, different concentration ranges of glutathione (from less to excess) could be rapidly detected by the two-stage ratiometric fluorescent probe 1 in 5 min. While, probe 2 shows single-stage ratiometric fluorescent detection to GSH (from green to reddish orange, I596nm/I535nm). Probes 1 and 2 exhibit excellent properties with sensitive, specific colorimetric response and ratiometric fluorescent response to glutathione over other sulfur nucleophiles. Application to cellular ratiometric fluorescence imaging indicated that the probes were highly responsive to intracellular glutathione.

A Phenylselenium-Substituted BODIPY Fluorescent Turn-off Probe for Fluorescence Imaging of Hydrogen Sulfide in Living Cells.

Herein a phenylselenium-substituted BODIPY (1) fluorescent turn-off sensor was developed for the purpose to achieve excellent selectivity and sensitivity for H2S detection based on the substitution reaction of the phenylselenide group at the 3-position with H2S. The excess addition of hydrogen sulfide promoted further substitution of the phenylselenide group at the 5-position of the probe and was accompanied by a further decrease in fluorescence emission intensity. Sensor 1 demonstrated remarkable performance with 49-fold red color fluorescence intensity decrease at longer excitation wavelength, a low detection limit (0.0025 µM), and specific fluorescent response toward H2S over anions, biothiols, and other amino acids in neutral media. It showed no obvious cell toxicity and good membrane permeability, which was well exploited for intracellular H2S detection and imaging through fluorescence microscopy imaging.

A fluorescence enhancement probe based on BODIPY for the discrimination of cysteine from homocysteine and glutathione.

Herein, a fluorescent probe BODIPY-based glyoxal hydrazone (BODIPY-GH) (1) for cysteine based on inhibiting of intramolecular charge transfer (ICT) quenching process upon reaction with the unsaturated aldehyde has been synthesized, which exhibits longer excitation wavelength, selective and sensitive colorimetric and fluorimetric response toward cysteine in natural media. The probe shows highly selectivity towards cysteine over homocysteine and glutathione as well as other amino acids with a significant fluorescence enhancement response within 15min In the presence of 50 equiv. of homocysteine, the emission increased slightly within 15min and completed in 2.5h to reach its maximum intensity. Therefore, the discrimination of cysteine from homocysteine and glutathione can be achieved through detection of probe 1. It shows low cytotoxicity and excellent membrane permeability toward living cells, which was successfully applied to detect and image intracellular cysteine effectively by confocal fluorescence imaging.