ATP-Activatable Photosensitizer Enables Dual Fluorescence Imaging and Targeted Photodynamic Therapy of Tumor

Targeted delivery of intracellular stimuli-activatable photosensitizers (PSs) into tumor cells to achieve selective imaging and on-demand photodynamic therapy (PDT) of tumors has provided a vital opportunity for precise cancer diagnosis and therapy. In this paper, we report a tumor targeting and adenosine triphosphate (ATP)-activatable nanophotosensitizer Apt-HyNP/BHQ₂ by modifying hybrid micellar nanoparticles with both nucleolin-targeting aptamer AS1411 and quencher BHQ₂-labeled ATP-binding aptamer BHQ₂-ATP-apt. We demonstrated that both of the fluorescence emissions at 555 and 627 nm were quenched by BHQ₂ in Apt-HyNP/BHQ₂, resulting in low PDT capacity. After selective entry into tumor cells through nucleolin-mediated endocytosis, the high concentration of intracellular ATP could bind to BHQ₂-ATP-apt and trigger Apt-HyNP/BHQ₂ dissociation, leading to turning “on” both fluorescence and PDT. The “off–on” fluorescence emissions at both 555 and 627 nm were successfully applied for dual color fluorescence imaging of endogenous ATP levels and real-time monitoring of intracellular activation of Apt-HyNP/BHQ₂ in tumor cells. Moreover, imaging-guided precise PDT of tumors in living mice was also demonstrated, allowing for selective ablation of tumors without obvious side effects. This study highlights the potential of using a combination of tumor-targeting and ATP-binding aptamers to design ATP-activatable PSs for both fluorescence imaging and imaging-guided PDT of tumors in vivo

Aggregation-Induced Electrochemiluminescence from a Cyclometalated Iridium(III) Complex

Aggregation-induced emission has been extensively found in organic compounds and metal complexes. In contrast, aggregation-induced electrochemiluminescence (AI-ECL) is rarely observed. Here, we employ two tridentate ligands [2,2′:6′,2″-terpyridine (tpy) and 1,3-bis­(1<i>H</i>-benzimidazol-2-yl)­benzene (bbbiH₃)] to construct a cyclometalated iridium­(III) complex, [Ir­(tpy)­(bbbi)] (<b>1</b>), showing strong AI-ECL. Its crystal structure indicates that neighboring [Ir­(tpy)­(bbbi)] molecules are connected through both π–π-stacking interactions and hydrogen bonds. These supramolecular interactions can facilitate the self-assembly of complex <b>1</b> into nanoparticles in an aqueous solution. The efficient restriction of molecular vibration in these nanoparticles leads to strong AI-ECL emission of complex <b>1</b>. In a dimethyl sulfoxide–water (H₂O) mixture with a gradual increase in the H₂O fraction from 20% to 98%, complex <b>1</b> showed a ∼39-fold increase in the electrochemiluminescence (ECL) intensity, which was ∼4.04 times as high as that of [Ru­(bpy)₃]²⁺ under the same experimental conditions. Moreover, the binding of bovine serum albumin to the nanoparticles of complex <b>1</b> can improve the ECL emission of this complex, facilitating the understanding of the mechanism of AI-ECL for future applications

Targeted Delivery of a γ‑Glutamyl Transpeptidase Activatable Near-Infrared-Fluorescent Probe for Selective Cancer Imaging

The noninvasive and specific detection of cancer cells in living subjects has been essential for the success of cancer diagnoses and treatments. Herein, we report a strategy of combining an α_vβ₃-integrin-receptor-targetable ligand, c-RGD, with the γ-glutamyl transpeptidase (GGT)-recognizable substrate, γ-glutamate (γ-Glu), to develop a tumor-targeting and GGT-activatable near-infrared (NIR)-fluorescent probe for the noninvasive imaging of tumors in living mice. We demonstrated that the probe’s fluorescence was off initially, but when the γ-Glu in the probe was specifically cleaved by GGT, the fluorescent product was released and could be selectively taken up by U87MG-tumor cells via α_vβ₃-receptor-mediated endocytosis. Remarkably, enhanced intracellular NIR fluorescence distributed mainly in the lysosomes was observed in the tumor cells only, showing that the probe was capable of differentiating the tumor cells from the GGT-positive, α_vβ₃-deficient normal cells. Moreover, the probe also showed a high selectivity for the real-time and noninvasive detection of GGT activity in xenograft U87MG tumors following iv administration. This study reveals the advantage of using a combination of receptor-mediated cell uptake and molecular-target-triggered activation to design molecular probes for improved cancer imaging, which could facilitate effective cancer diagnoses

Lysosome-Targeting Fluorogenic Probe for Cathepsin B Imaging in Living Cells

Cathepsin B (CTB) is a lysosomal protease which has been recognized as a promising biomarker for many malignant tumors, and accurate detection of its activity is important in early diagnosis of cancers and predicting metastasis. Herein, we reported a lysosome-targeting fluorogenic small-molecule probe for fluorescence imaging of lysosomal CTB in living cancer cells by incorporating a CTB-recognitive peptide substrate Cbz-Lys-Lys-<i>p</i>-aminobenzyl alcohol (Cbz-Lys-Lys-PABA) and a lysosome locating group morpholine. We demonstrated that the probe could be efficiently activated by CTB to generate ∼73-fold enhancement in fluorescence under acidic lysosomal environment (pH 4.5–6.0), allowing for high sensitivity and specificity to detect CTB. Fluorescence imaging results showed selective accumulation and fluorescence turn-on in the lysosomes of cancer cells, which were capable of reporting on lysosomal CTB activity in cancer cells and normal tissue cells. This study highlights the potential of using a lysosome-targeting group to design a sensitive and specific fluorogenic probe for fluorescence imaging of lysosomal CTB in living cells

Surface-Roughened SERS-Active Single Silver Nanowire for Simultaneous Detection of Intracellular and Extracellular pHs

The simultaneous and accurate detection of intracellular pH (pHi) and extracellular pH (pHe) is essential for studying the complex physiological activities of cancer cells and exploring pH-related therapeutic mechanisms. Here, we developed a super-long silver nanowire-based surface-enhanced Raman scattering (SERS) detection strategy for simultaneous sensing of pHi and pHe. A surface-roughened silver nanowire (AgNW) with a high aspect ratio is prepared at a nanoelectrode tip using a Cu-mediated oxidation process, which is then modified by pH-sensitive 4-mercaptobenzoic acid (4-MBA) to form 4-MBA@AgNW as a pH sensing probe. With the assistance of a 4D microcontroller, 4-MBA@AgNW is efficient in simultaneously detecting pHi and pHe in both 2D and 3D culture cancer cells by SERS, with minimal invasiveness, high sensitivity, and spatial resolution. Further investigation proves that the surface-roughened single AgNW can also be used in monitoring the dynamic variation of pHi and pHe of cancer cells upon stimulation with anticancer drugs or under a hypoxic environment

Activatable QD-Based Near-Infrared Fluorescence Probe for Sensitive Detection and Imaging of DNA

Accurate detection of DNA is essential for the precise diagnosis of diseases. Here we report an activatable near-infrared (NIR) fluorescence nanoprobe (QD-Al-GFLX) composed of NIR quantum dots (QDs) and Al­(III)-gatifloxacin (Al-GFLX) complexes for the sensitive detection of double-stranded DNA (dsDNA) both in aqueous solution and in living cells. We demonstrated that the initial strong NIR fluorescence of QDs in QD-Al-GFLX was quenched by the Al-GFLX complex via a photoinduced electron transfer (PET) mechanism. Upon interaction with dsDNA, the high binding affinity between dsDNA and Al-GFLX complex could trigger QD-Al-GFLX dissociation, which could eliminate the PET process, resulting in significant enhancement of NIR fluorescence. QD-Al-GFLX was sensitive and specific to detect dsDNA in aqueous solution, with a detection limit of 6.83 ng/mL. The subsequent fluorescence imaging revealed that QD-Al-GFLX holds a high ability to enter into live cells, generating strong NIR fluorescence capable of reporting on dsDNA levels. This study highlighted the potential of using QD-Al-GFLX nanoprobe for the real-time detection and imaging of dsDNA in living cells

Surface-Roughened SERS-Active Single Silver Nanowire for Simultaneous Detection of Intracellular and Extracellular pHs

The simultaneous and accurate detection of intracellular pH (pHi) and extracellular pH (pHe) is essential for studying the complex physiological activities of cancer cells and exploring pH-related therapeutic mechanisms. Here, we developed a super-long silver nanowire-based surface-enhanced Raman scattering (SERS) detection strategy for simultaneous sensing of pHi and pHe. A surface-roughened silver nanowire (AgNW) with a high aspect ratio is prepared at a nanoelectrode tip using a Cu-mediated oxidation process, which is then modified by pH-sensitive 4-mercaptobenzoic acid (4-MBA) to form 4-MBA@AgNW as a pH sensing probe. With the assistance of a 4D microcontroller, 4-MBA@AgNW is efficient in simultaneously detecting pHi and pHe in both 2D and 3D culture cancer cells by SERS, with minimal invasiveness, high sensitivity, and spatial resolution. Further investigation proves that the surface-roughened single AgNW can also be used in monitoring the dynamic variation of pHi and pHe of cancer cells upon stimulation with anticancer drugs or under a hypoxic environment

Surface-Roughened SERS-Active Single Silver Nanowire for Simultaneous Detection of Intracellular and Extracellular pHs

The simultaneous and accurate detection of intracellular pH (pHi) and extracellular pH (pHe) is essential for studying the complex physiological activities of cancer cells and exploring pH-related therapeutic mechanisms. Here, we developed a super-long silver nanowire-based surface-enhanced Raman scattering (SERS) detection strategy for simultaneous sensing of pHi and pHe. A surface-roughened silver nanowire (AgNW) with a high aspect ratio is prepared at a nanoelectrode tip using a Cu-mediated oxidation process, which is then modified by pH-sensitive 4-mercaptobenzoic acid (4-MBA) to form 4-MBA@AgNW as a pH sensing probe. With the assistance of a 4D microcontroller, 4-MBA@AgNW is efficient in simultaneously detecting pHi and pHe in both 2D and 3D culture cancer cells by SERS, with minimal invasiveness, high sensitivity, and spatial resolution. Further investigation proves that the surface-roughened single AgNW can also be used in monitoring the dynamic variation of pHi and pHe of cancer cells upon stimulation with anticancer drugs or under a hypoxic environment

Magnetic Resonance Imaging of Stem Cell Apoptosis in Arthritic Joints with a Caspase Activatable Contrast Agent

About 43 million individuals in the U.S. encounter cartilage injuries due to trauma or osteoarthritis, leading to joint pain and functional disability. Matrix-associated stem cell implants (MASI) represent a promising approach for repair of cartilage defects. However, limited survival of MASI creates a significant bottleneck for successful cartilage regeneration outcomes and functional reconstitution. We report an approach for noninvasive detection of stem cell apoptosis with magnetic resonance imaging (MRI), based on a caspase-3-sensitive nanoaggregation MRI probe (C-SNAM). C-SNAM self-assembles into nanoparticles after hydrolysis by caspase-3, leading to 90% amplification of ¹H MR signal and prolonged <i>in vivo</i> retention. Following intra-articular injection, C-SNAM causes significant MR signal enhancement in apoptotic MASI compared to viable MASI. Our results indicate that C-SNAM functions as an imaging probe for stem cell apoptosis in MASI. This concept could be applied to a broad range of cell transplants and target sites

Redox-Mediated Disassembly to Build Activatable Trimodal Probe for Molecular Imaging of Biothiols

Activatable multimodal probes that show enhancement of multiplex imaging signals upon interaction with their specific molecular target have become powerful tools for rapid and precise imaging of biological processes. Herein, we report a stimuli-responsive disassembly approach to construct a redox-activatable fluorescence/¹⁹F-MRS/¹H-MRI triple-functional probe <b>1</b>. The small molecule probe <b>1</b> itself has a high propensity to self-assemble into nanoparticles with quenched fluorescence, attenuated ¹⁹F-MRS signal, and high ¹H-MRI contrast. Biothiols that are abundant in reducing biological environment were able to cleave the disulfide bond in probe <b>1</b> to induce disassembly of the nanoparticles and lead to fluorescence activation (∼70-fold), ¹⁹F-MRS signal amplification (∼30-fold) and significant <i>r</i>₁ relaxivity reduction (∼68% at 0.5 T). Molecular imaging of reducing environment in live cells and <i>in vivo</i> was realized using probe <b>1</b>. This approach could facilitate the development of other stimuli-responsive trimodal probes for molecular imaging