Adhesion molecule-targeted magnetic particle imaging nanoprobe for visualization of inflammation in acute lung injury.

PURPOSE: Uncontrolled intra-alveolar inflammation is a central pathogenic feature, and its severity translates into a valid prognostic indicator of acute lung injury (ALI). Unfortunately, current clinical imaging approaches are unsuitable for visualizing and quantifying intra-alveolar inflammation. This study aimed to construct a small-sized vascular cell adhesion molecule-1 (VCAM-1)-targeted magnetic particle imaging (MPI) nanoprobe (ESPVPN) to visualize and accurately quantify intra-alveolar inflammation at the molecular level. METHODS: ESPVPN was engineered by conjugating a peptide (VHPKQHRGGSK(Cy7)GC) onto a polydopamine-functionalized superparamagnetic iron oxide core. The MPI performance, targeting, and biosafety of the ESPVPN were characterized. VCAM-1 expression in HUVECs and mouse models was evaluated by western blot. The degree of inflammation and distribution of VCAM-1 in the lungs were assessed using histopathology. The expression of pro-inflammatory markers and VCAM-1 in lung tissue lysates was measured using ELISA. After intravenous administration of ESPVPN, MPI and CT imaging were used to analyze the distribution of ESPVPN in the lungs of the LPS-induced ALI models. RESULTS: The small-sized (~10 nm) ESPVPN exhibited superior MPI performance compared to commercial MagImaging® and Vivotrax, and ESPVPN had effective targeting and biosafety. VCAM-1 was highly expressed in LPS-induced ALI mice. VCAM-1 expression was positively correlated with the LPS-induced dose (R = 0.9381). The in vivo MPI signal showed positive correlations with both VCAM-1 expression (R = 0.9186) and representative pro-inflammatory markers (MPO, TNF-α, IL-6, IL-8, and IL-1ß, R > 0.7). CONCLUSION: ESPVPN effectively targeted inflammatory lungs and combined the advantages of MPI quantitative imaging to visualize and evaluate the degree of ALI inflammation.

Biomimetic manganese-eumelanin nanocomposites for combined hyperthermia-immunotherapy against prostate cancer.

Pro-tumoral and immunosuppressive M2-like tumor-associated macrophages (TAMs) contribute to tumor progression, recurrence and distal metastasis. However, current TAMs-modulating therapeutic strategies often encounter challenges including insufficient immune activation, weak antigen presentation ability and unsatisfactory antitumor immune performance. Herein, cyclic RGD peptide functionalized and manganese doped eumelanin-like nanocomposites (RMnMels) are reported for combined hyperthermia-immunotherapy against PC3 prostate cancer. The RMnMels could promote M2-to-M1 macrophage repolarization via scavenging multiple reactive oxygen species and remodeling the immunosuppressive tumor microenvironment. Following near-infrared light irradiation, RMnMels-mediated thermal ablation not only could destroy tumor cells directly, but also elicit the release of damage associated molecular patterns and tumor-associated antigens, provoking robust tumor immunogenicity and strong antitumor immune responses. The results showed that RMnMels could effectively scavenge reactive oxygen species and promote M2-to-M1 macrophage repolarization both in vitro and in vivo. Synergistically enhanced anti-tumor therapeutic efficacy was achieved following single administration of RMnMels plus single round of laser irradiation, evidenced by decreased primary tumor sizes and decreased number of distant liver metastatic nodules. The as-developed RMnMels may represent a simple and high-performance therapeutic nanoplatform for immunomodulation and enhanced antitumor immune responses.

Colourful fluorescence-based carbon dots for tumour imaging-guided nanosurgery.

Fluorescent-intraoperative navigation is a visual technique that allows surgeons to accurately distinguish malignant and normal tissues during surgery. It has the advantages of immediacy, high resolution, and high specificity. However, a single fluorescent source cannot provide sufficient surgical information. Multicolour carbon dots (CDs) are more suitable since they provide outstanding water solubility, photostability, and multicolour-fluorescence imaging. Here, we prepared an optical probe with CD-based multicolour-fluorescence imaging via a hydrothermal method. CDs can be endocytosed by tumour cells, and after intravenous injection, they can effectively accumulate at the tumour site. In a pancreatic cancer mouse model, we demonstrated the multicolour-fluorescence imaging capabilities of CDs, which aided the accurate resection of tumours under fluorescent-intraoperative navigation. Stereoscopic fluorescence microscopy imaging and H&E staining proved that the removed tissue belonged to the pancreatic tumour. This study emphasizes the potential of CDs for fluorescence-guided intraoperative resection and expands the application of CDs in biological fields.

Fluorine-Doped Carbon Dots with Intrinsic Nucleus-Targeting Ability for Drug and Dye Delivery.

A new type of fluorine-doped carbon dots (FCDs) with the nucleus-targeting capability was prepared and utilized as a promising candidate for drug and dye delivery. Doxorubicin (DOX) and boron dipyrromethene (BODIPY) was used as a model drug and dye, respectively, to construct FCD-DOX and FCD-BODIPY nanocomposites via coassembly with FCDs. The results demonstrate that FCDs can remarkably increase the cellular uptake and delivery of DOX and BODIPY. This work developed a convenient strategy to construct CDs-based nanohybrids for nucleus-targeted bioimaging and cancer treatment.

Cell-Penetrating Peptides Transport Noncovalently Linked Thermally Activated Delayed Fluorescence Nanoparticles for Time-Resolved Luminescence Imaging.

Luminescent probes and nanoparticles (NPs) with long excited state lifetimes are essential for time-resolved biological imaging. Generally, cell membranes are physiological barriers that could prevent the uptake of many unnatural compounds. It is still a big challenge to prepare biocompatible imaging agents with high cytomembrane permeability, especially for nonmetallic NPs with long-lived luminescence. Herein, an amphiphilic cell-penetrating peptide, F6G6(rR)3R2, was designed to transport hydrophobic fluorophores across cellular barriers. Three classical thermally activated delayed fluorescence (TADF) molecules, 4CzIPN, NAI-DPAC, and BTZ-DMAC, could self-assemble into well-dispersed NPs with F6G6(rR)3R2 in aqueous solution. These NPs showed low cytotoxicity and could penetrate membranes easily. Moreover, long-lived TADF enabled them to be used in time-resolved luminescence imaging in oxygenic environments. These findings greatly expanded the applications of cell-penetrating peptides for delivery of molecules and NPs by only noncovalent interactions, which were more flexible and easier than covalent modifications.

Dual-Phosphorescent Iridium(III) Complexes Extending Oxygen Sensing from Hypoxia to Hyperoxia.

Hypoxia and hyperoxia, referring to states of biological tissues in which oxygen supply is in sufficient and excessive, respectively, are often pathological conditions. Many luminescent oxygen probes have been developed for imaging intracellular and in vivo hypoxia, but their sensitivity toward hyperoxia becomes very low. Here we report a series of iridium(III) complexes in which limited internal conversion between two excited states results in dual phosphorescence from two different excited states upon excitation at a single wavelength. Structural manipulation of the complexes allows rational tuning of the dual-phosphorescence properties and the spectral profile response of the complexes toward oxygen. By manipulating the efficiency of internal conversion between the two emissive states, we obtained a complex exhibiting naked-eye distinguishable green, orange, and red emission in aqueous buffer solution under an atmosphere of N2, air, and O2, respectively. This complex is used for intracellular and in vivo oxygen sensing not only in the hypoxic region but also in normoxic and hyperoxic intervals. To the best of our knowledge, this is the first example of using a molecular probe for simultaneous bioimaging of hypoxia and hyperoxia.

First magnetic particle imaging to assess pulmonary vascular leakage in vivo in the acutely injured and fibrotic lung.

Increased pulmonary vascular permeability is a characteristic feature of lung injury. However, there are no established methods that allow the three-dimensional visualization and quantification of pulmonary vascular permeability in vivo. Evans blue extravasation test and total protein test of bronchoalveolar lavage fluid (BALF) are permeability assays commonly used in research settings. However, they lack the ability to identify the spatial and temporal heterogeneity of endothelial barrier disruption, which is typical in lung injuries. Magnetic resonance (MR) and near-infrared (NIR) imaging have been proposed to image pulmonary permeability, but suffer from limited sensitivity and penetration depth, respectively. In this study, we report the first use of magnetic particle imaging (MPI) to assess pulmonary vascular leakage noninvasively in vivo in mice. A dextran-coated superparamagnetic iron oxide (SPIO), synomag®, was employed as the imaging tracer, and pulmonary SPIO extravasation was imaged and quantified to evaluate the vascular leakage. Animal models of acute lung injury and pulmonary fibrosis (PF) were used to validate the proposed method. MPI sensitively detected the SPIO extravasation in both acutely injured and fibrotic lungs in vivo, which was confirmed by ex vivo imaging and Prussian blue staining. Moreover, 3D MPI illustrated the spatial heterogeneity of vascular leakage, which correlated well with CT findings. Based on the in vivo 3D MPI images, we defined the SPIO extravasation index (SEI) to quantify the vascular leakage. A significant increase in SEI was observed in the injured lungs, in consistent with the results obtained via ex vivo permeability assays. Overall, our results demonstrate that 3D quantitative MPI serves as a useful tool to examine pulmonary vascular integrity in vivo, which shows promise for future clinical translation.

Magnetic Particle Imaging-Guided Hyperthermia for Precise Treatment of Cancer: Review, Challenges, and Prospects.

Magnetic particle imaging (MPI) is a novel quantitative imaging technique using the nonlinear magnetization behavior of magnetic nanoparticles (MNPs) to determine their local concentration. Magnetic fluid hyperthermia (MFH) is a promising non-invasive therapy using the heating effects of MNPs. MPI-MFH is expected to enable real-time MPI guidance, localized MFH, and non-invasive temperature monitoring, which shows great potential for precise treatment of cancer. In this review, we introduce the fundamentals of MPI and MFH and their applications in the treatment of cancer. Also, we discuss the challenges and prospects of MPI-MFH.

Characterization and anti-aging effects of polysaccharide from Gomphus clavatus Gray.

In this study, a highly branched polysaccharide (GPF, 112.0 kDa) was isolated and purified from Gomphus clavatus Gray fruiting bodies. GPF was primarily composed of mannose, galactose, arabinose, xylose, and glucose at a molar ratio of 3.2:1.9:1.6:1.2:1.0. GPF was a highly branched heteropolysaccharide composed of 13 glucosidic bonds, with a degree of branching (DB) of 48.85 %. GPF exhibited anti-aging activity in vivo, significantly increased antioxidant enzymes activities (SOD, CAT and GSH-Px), improved total antioxidant capability (T-AOC) and decreased MDA level in the serum and brain of d-Gal induced aging mice. Behavioral experiments showed that GPF effectively improved learning and memory deficits in d-Gal induced aging mice. Mechanistic studies indicated that GPF could activate AMPK by increasing AMPK phosphorylation and upregulating SIRT1 and PGC-1α expression. These findings suggest that GPF has significant potential as a natural candidate to slow down aging and prevent aging-related diseases.

PGNet: Projection generative network for sparse-view reconstruction of projection-based magnetic particle imaging.

BACKGROUND: Magnetic particle imaging (MPI) is a novel tomographic imaging modality that scans the distribution of superparamagnetic iron oxide nanoparticles. However, it is time-consuming to scan multiview two-dimensional (2D) projections for three-dimensional (3D) reconstruction in projection MPI, such as computed tomography (CT). An intuitive idea is to use the sparse-view projections for reconstruction to improve the temporal resolution. Tremendous progress has been made toward addressing the sparse-view problem in CT, because of the availability of large data sets. For the novel tomography of MPI, to the best of our knowledge, studies on the sparse-view problem have not yet been reported. PURPOSE: The acquisition of multiview projections for 3D MPI imaging is time-consuming. Our goal is to only acquire sparse-view projections for reconstruction to improve the 3D imaging temporal resolution of projection MPI. METHODS: We propose to address the sparse-view problem in projection MPI by generating novel projections. The data set we constructed consists of three parts: simulation data set (including 3000 3D data), four phantoms data, and an in vivo mouse data. The simulation data set is used to train and validate the network, and the phantoms and in vivo mouse data are used to test the network. When the number of novel generated projections meets the requirements of filtered back projection, the streaking artifacts will be absent from MPI tomographic imaging. Specifically, we propose a projection generative network (PGNet), that combines an attention mechanism, adversarial training strategy, and a fusion loss function and can generate novel projections based on sparse-view real projections. To the best of our knowledge, we are the first to propose a deep learning method to attempt to overcome the sparse-view problem in projection MPI. RESULTS: We compare our method with several sparse-view methods on phantoms and in vivo mouse data and validate the advantages and effectiveness of our proposed PGNet. Our proposed PGNet enables the 3D imaging temporal resolution of projection MPI to be improved by 6.6 times, while significantly suppressing the streaking artifacts. CONCLUSION: We proposed a deep learning method operated in projection domain to address the sparse-view reconstruction of MPI, and the data scarcity problem in projection MPI reconstruction is alleviated by constructing a sparse-dense simulated projection data set. By our proposed method, the number of acquisitions of real projections can be reduced. The advantage of our method is that it prevents the generation of streaking artifacts at the source. Our proposed sparse-view reconstruction method has great potential for application to time-sensitive in vivo 3D MPI imaging.

Deep Penetrating and Sensitive Targeted Magnetic Particle Imaging and Photothermal Therapy of Early-Stage Glioblastoma Based on a Biomimetic Nanoplatform.

Early diagnosis can effectively improve the survival of glioblastoma multiforme (GBM). A specific imaging technique that is simultaneously deep penetrating and sensitive to small tissue changes is desired to identify GBM. Due to its excellent features in signal contrast, detection sensitivity, and none or little attenuation in tissue, magnetic particle imaging (MPI) possesses great potential in cancer diagnosis, especially when the imaging modality is equipped with specifically targeted nanoprobes. However, when gliomas are small, the blood-brain barrier (BBB) is complete and prevents nanoprobes from entering the brain, which negates the theranostic effect. This study proposes a biomimetic nanoplatform that assist the MPI tracers in breaking through the BBB and then demonstrate a targeted and sensitive diagnosis of GBM. Afterward, the photothermal therapy and immune regulation show an excellent therapeutic effect on the GBM. It is experimentally confirmed that the MPI signal does not decay with tissue depth and shows excellent sensitivity for thousands-cells. Only small animals are conducted in this study due to the limitations of the current commercial MPI scanner, however, this research theoretically enables large animal and human studies, which encourages a promising pathway toward the noninvasive diagnosis of early-stage GBM in clinics.

Cross-domain knowledge transfer based parallel-cascaded multi-scale attention network for limited view reconstruction in projection magnetic particle imaging.

Projection magnetic particle imaging (MPI) can significantly improve the temporal resolution of three-dimensional (3D) imaging compared to that using traditional point by point scanning. However, the dense view of projections required for tomographic reconstruction limits the scope of temporal resolution optimization. The solution to this problem in computed tomography (CT) is using limited view projections (sparse view or limited angle) for reconstruction, which can be divided into: completing the limited view sinogram and image post-processing for streaking artifacts caused by insufficient projections. Benefiting from large-scale CT datasets, both categories of deep learning-based methods have achieved tremendous progress; yet, there is a data scarcity limitation in MPI. We propose a cross-domain knowledge transfer learning strategy that can transfer the prior knowledge of the limited view learned by the model in CT to MPI, which can help reduce the network requirements for real MPI data. In addition, the size of the imaging target affects the scale of the streaking artifacts caused by insufficient projections. Therefore, we propose a parallel-cascaded multi-scale attention module that allows the network to adaptively identify streaking artifacts at different scales. The proposed method was evaluated on real phantom and in vivo mouse data, and it significantly outperformed several advanced limited view methods. The streaking artifacts caused by an insufficient number of projections can be overcome using the proposed method.

Chiral Carbon Dots-Enzyme Nanoreactors with Enhanced Catalytic Activity for Cancer Therapy.

As a class of functional proteins, enzymes possess inherent insignificant features, for instance, mediocre stability and membrane impermeability and reduced enzymatic activity after modification, which partly limit their biomedical applications. Thus, it is indispensable to exploit robust nanoreactors with high enzymatic activity and good stability and cell permeability. Here, the chiral carbon dots (CDs)-glucose oxidase (GOx) nanoreactors named LGOx and DGOx were constructed by the coassembly of GOx with L/D-CDs, respectively. L/DGOx can significantly enhance the activity of GOx and improve the efficient delivery of GOx to cancer cells. Moreover, these nanoreactors can generate hydrogen peroxide to efficaciously kill cancer cells and restrain tumor growth, and DGOx exhibits higher enzymatic activity than LGOx. According to our understanding, this is the first report about utilizing chiral CDs as vectors to construct effective CDs-enzyme nanohybrids for cancer therapy, which is envisioned to be a versatile strategy for multitudinous biomedical applications.

Dual-Emissive Phosphorescent Polymer Probe for Accurate Temperature Sensing in Living Cells and Zebrafish Using Ratiometric and Phosphorescence Lifetime Imaging Microscopy.

Temperature plays an important part in many biochemical processes. Accurate diagnosis and proper treatment usually depend on precise measurement of temperature. In this work, a dual-emissive phosphorescent polymer temperature probe, composed of iridium(III) complexes as temperature sensitive unit with phosphorescence lifetime of ∼500 ns and europium(III) complexes as reference unit with lifetime of ∼400 µs, has been rationally designed and synthesized. Upon the increase of the temperature, the luminescence intensity from the iridium(III) complexes is enhanced, while that from the europium(III) complexes remains unchanged, which makes it possible for the ratiometric detection of temperature. Furthermore, the polymer also displays a significant change in emission lifetime accompanied by the temperature variation. By utilizing the laser scanning confocal microscope and time-resolved luminescence imaging systems, ratiometric and time-resolved luminescence imaging in Hela cells and zebrafish have been carried out. Notably, the intensity ratio and long-lifetime-based imaging can offer higher sensitivity, decrease the detection limit, and minimize the background interference from biosamples.

A novel phosphorescent iridium(iii) complex bearing a donor-acceptor-type o-carboranylated ligand for endocellular hypoxia imaging.

The structure-property relationship of carborane-embedded cationic iridium(iii) complexes was investigated in this work, especially with donor-acceptor-type ligands. Firstly, an efficient synthetic approach for the new donor-acceptor-type ligands (2 and 4) was developed. By using these ligands, novel iridium(iii) complexes II and IV were prepared. Complex IV shows about a 92 nm red-shift in solution (ΦP = 0.13 in ethanol), and the emission color has an obvious change from green to red when compared with the Model complex. In addition, the photophysical characteristics of complex IV are quite sensitive to the oxygen level in living cells, based on which endocellular hypoxia imaging of complex IV has been realized by the phosphorescence lifetime imaging microscopy (PLIM) technology. Besides the experimental studies, density functional theory (DFT) and time dependent DFT (TD-DFT) calculations have been successfully applied to investigate the excited-state electronic structures of carborane-modified iridium complexes.

Luminescent gold nanocluster-based sensing platform for accurate H2S detection in vitro and in vivo with improved anti-interference.

Gold nanoclusters (Au NCs) are promising luminescent nanomaterials due to their outstanding optical properties. However, their relatively low quantum yields and environment-dependent photoluminescence properties have limited their biological applications. To address these problems, we developed a novel strategy to prepare chitosan oligosaccharide lactate (Chi)-functionalized Au NCs (Au NCs@Chi), which exhibited emission with enhanced quantum yield and elongated emission lifetime as compared to the Au NCs, as well as exhibited environment-independent photoluminescence properties. In addition, utilizing the free amino groups of Chi onto Au NCs@Chi, we designed a FRET-based sensing platform for the detection of hydrogen sulfide (H2S). The Au NCs and the specific H2S-sensitive merocyanine compound were respectively employed as an energy donor and acceptor in the platform. The addition of H2S induced changes in the emission profile and luminescence lifetime of the platform with high sensitivity and selectivity. Utilization of the platform was demonstrated to detect exogenous and endogenous H2S in vitro and in vivo through wavelength-ratiometric and time-resolved luminescence imaging (TLI). Compared to previously reported luminescent molecules, the platform was less affected by experimental conditions and showed minimized autofluorescence interference and improved accuracy of detection.

Hyperbranched Phosphorescent Conjugated Polymer Dots with Iridium(III) Complex as the Core for Hypoxia Imaging and Photodynamic Therapy.

Real-time monitoring of the contents of molecular oxygen (O2) in tumor cells is of great significance in early diagnosis of cancer. At the same time, the photodynamic therapy (PDT) could be realized by highly toxic singlet oxygen (1O2) generated in situ during the O2 sensing, making it one of the most promising methods for cancer therapy. Herein, the iridium(III) complex cored hyperbranched phosphorescent conjugated polymer dots with the negative charges for hypoxia imaging and highly efficient PDT was rationally designed and synthesized. The incomplete energy transfer between the polyfluorene and the iridium(III) complexes realized the ratiometric sensing of O2 for the accurate measurements. Furthermore, the O2-dependent emission lifetimes are also used in photoluminescence lifetime imaging and time-gated luminescence imaging for eliminating the autofluorescence remarkably to enhance the signal-to-noise ratio of imaging. Notably, the polymer dots designed could generate the 1O2 effectively in aqueous solution, and the image-guided PDT of the cancer cells was successfully realized and investigated in detail by confocal laser scanning microscope. To the best of our knowledge, this represents the first example of the iridium(III) complex cored hyperbranched conjugated polymer dots with the negative charges for both hypoxia imaging and PDT of cancer cells simultaneously.