The Nanoassembly of an Intrinsically Cytotoxic Near-Infrared Dye for Multifunctionally Synergistic Theranostics.

The combination of diagnostic and therapeutic functions in a single theranostic nanoagent generally requires the integration of multi-ingredients. Herein, a cytotoxic near-infrared (NIR) dye (IR-797) and its nanoassembly are reported for multifunctional cancer theranostics. The hydrophobic IR-797 molecules are self-assembled into nanoparticles, which are further modified with an amphiphilic polymer (C18PMH-PEG5000) on the surface. The prepared PEG-IR-797 nanoparticles (PEG-IR-797 NPs) possess inherent cytotoxicity from the IR-797 dye and work as a chemotherapeutic drug which induces apoptosis of cancer cells. The IR-797 NPs are found to have an ultrahigh mass extinction coefficient (444.3 L g-1 cm-1 at 797 nm and 385.9 L g-1 cm-1 at 808 nm) beyond all reported organic nanomaterials (<40 L g-1 cm-1 ) for superior photothermal therapy (PTT). In addition, IR-797 shows some aggregation-induced-emission (AIE) properties. Combining the merits of good NIR absorption, high photothermal energy conversion efficiency, and AIE, makes the PEG-IR-797 NPs useful for multimodal NIR AIE fluorescence, photoacoustic, and thermal imaging-guided therapy. The research exhibits the possibility of using a single ingredient and entity to perform multimodal NIR fluorescence, photoacoustic, and thermal imaging-guided chemo-/photothermal combination therapy, which may trigger wide interest from the fields of nanomedicine and medicinal chemistry to explore multifunctional theranostic organic molecules.

Correction: An, F., et al. A Conjugate of Pentamethine Cyanine and 18F as a Positron Emission Tomography/Near-Infrared Fluorescence Probe for Multimodality Tumor Imaging. Int. J. Mol. Sci. 2017, 18, 1214.

The authors wish to make the following corrections to this paper [...].

The impact of light irradiation timing on the efficacy of nanoformula-based photo/chemo combination therapy.

Photo/chemo combination therapy has been demonstrated to be a generally more powerful strategy for treating cancers than a single treatment modality. However, it is unknown whether the timing of light irradiation has any impact on therapeutic efficacy. We designed a carrier-free and self-monitoring nanodrug to monitor the entire dual-drug release profile and determined the impact of photodynamic therapy (PDT) at different time points. The designed nanodrug consists of the chemotherapeutic doxorubicin (DOX) and the photosensitizer pheophorbide A (PhA). The drugs form a fluorescence resonance energy transfer (FRET) pair (DOX transferring energy to PhA) when present at a precise ratio in the combination nanodrug. Due to the FRET effect, the DOX-PhA nanoparticles (NPs) show PhA fluorescence in a normal pH environment (such as cytoplasm). However, the FRET effect is lost when the NPs are disassembled in an acidic environment (such as lysosomes), and the DOX fluorescence is recovered. By real-time fluorescence variation monitoring, we determined the key time points when the drugs reached various subcellular locations, which helped us to determine the PDT-triggering time points and investigate the impact on the therapeutic effect in the combination therapy. Furthermore, the PDT was triggered at these established time points both in vitro and in vivo, which revealed that the best PDT-triggering time point in the combination therapy was achieved after nuclear entry of DOX. The study suggests that the optimization of combination therapy, not only photo/chemo but also chemo/chemo combination therapy, may require not only a controlled drug ratio but also a controlled drug release profile and target arrival time.

Strategies for Preparing Albumin-based Nanoparticles for Multifunctional Bioimaging and Drug Delivery.

Biosafety is the primary concern in clinical translation of nanomedicine. As an intrinsic ingredient of human blood without immunogenicity and encouraged by its successful clinical application in Abraxane, albumin has been regarded as a promising material to produce nanoparticles for bioimaging and drug delivery. The strategies for synthesizing albumin-based nanoparticles could be generally categorized into five classes: template, nanocarrier, scaffold, stabilizer and albumin-polymer conjugate. This review introduces approaches utilizing albumin in the preparation of nanoparticles and thereby provides scientists with knowledge of goal-driven design on albumin-based nanomedicine.

A Conjugate of Pentamethine Cyanine and 18F as a Positron Emission Tomography/Near-Infrared Fluorescence Probe for Multimodality Tumor Imaging.

The novel synthesis of a dual-modality, pentamethine cyanine (Cy5) fluorescent, 18F positron emission tomography (PET) imaging probe is reported. The probe shows a large extinction coefficient and large quantum yield in the biologically transparent, near-infrared window (650-900 nm) for in vivo fluorescent imaging. This fluorophore bears the isotope, 18F, giving a 18F-PET/near-infrared fluorescent (NIRF), bi-modal imaging probe, that combines the long-term stability of NIRF and the unlimited penetration depth of PET imaging. The bi-modal probe is labeled with 18F in a quick, one-step reaction, which is important in working with the rapid decay of 18F. The bi-modal probe bears a free carboxyl group, highlighting a PET/NIRF synthon that can be conjugated onto many advanced biomolecules for biomarker-specific in vivo dual-modal PET/NIR tumor imaging, confocal histology, and utility in multi-fluorophore, fluorescence-guided surgery. Its potential in vivo biocompatibility is explored in a quick proof-of-principal in vivo study. The dye is delivered to A549 xenograft flank-tumors to generate PET and NIRF signals at the tumor site. The tumor distribution is confirmed in ex vivo gamma counting and imaging. Pentamethine cyanine (Cy5) has the ability to preferentially accumulate in tumor xenografts. We substitute the PET/NIRF probe for Cy5, and explore this phenomenon.

18F-positron-emitting/fluorescent labeled erythrocytes allow imaging of internal hemorrhage in a murine intracranial hemorrhage model.

An agent for visualizing cells by positron emission tomography is described and used to label red blood cells. The labeled red blood cells are injected systemically so that intracranial hemorrhage can be visualized by positron emission tomography (PET). Red blood cells are labeled with 0.3 µg of a positron-emitting, fluorescent multimodal imaging probe, and used to non-invasively image cryolesion induced intracranial hemorrhage in a murine model (BALB/c, 2.36 × 108 cells, 100 µCi, <4 mm hemorrhage). Intracranial hemorrhage is confirmed by histology, fluorescence, bright-field, and PET ex vivo imaging. The low required activity, minimal mass, and high resolution of this technique make this strategy an attractive alternative for imaging intracranial hemorrhage. PET is one solution to a spectrum of issues that complicate single photon emission computed tomography (SPECT). For this reason, this application serves as a PET alternative to [99mTc]-agents, and SPECT technology that is used in 2 million annual medical procedures. PET contrast is also superior to gadolinium and iodide contrast angiography for its lack of clinical contraindications.

Dual PET and Near-Infrared Fluorescence Imaging Probes as Tools for Imaging in Oncology.

OBJECTIVE: The purpose of this article is to summarize advances in PET fluorescence resolution, agent design, and preclinical imaging that make a growing case for clinical PET fluorescence imaging. CONCLUSION: Existing SPECT, PET, fluorescence, and MRI contrast imaging techniques are already deeply integrated into the management of cancer, from initial diagnosis to the observation and management of metastases. Combined positron-emitting fluorescent contrast agents can convey new or substantial benefits that improve on these proven clinical contrast agents.

Self-carried curcumin nanoparticles for in vitro and in vivo cancer therapy with real-time monitoring of drug release.

The use of different nanocarriers for delivering hydrophobic pharmaceutical agents to tumor sites has garnered major attention. Despite the merits of these nanocarriers, further studies are needed to improve their drug loading capacities (which are typically <10%) and reduce their potential systemic toxicity. Therefore, the development of alternative self-carried nanodrug delivery strategies without using inert carriers is highly desirable. In this study, we developed a self-carried curcumin (Cur) nanodrug for highly effective cancer therapy in vitro and in vivo with real-time monitoring of drug release. With a biocompatible C18PMH-PEG functionalization, the Cur nanoparticles (NPs) showed excellent dispersibility and outstanding stability in physiological environments with drug loading capacities >78 wt%. Both confocal microscopy and flow cytometry confirmed the cellular fluorescence "OFF-ON" activation and real-time monitoring of the Cur molecule release. In vitro and in vivo experiments clearly show that the therapeutic efficacy of the PEGylated Cur NPs is considerably better than that of free Cur. This self-carried strategy with real-time monitoring of drug release may open a new way for simultaneous cancer therapy and monitoring.

Preparation and size control of sub-100 nm pure nanodrugs.

Pure nanodrugs (PNDs), nanoparticles consisting entirely of drug molecules, have been considered as promising candidates for next-generation nanodrugs. However, the traditional preparation method via reprecipitation faces critical challenges including low production rates, relatively large particle sizes, and batch-to-batch variations. Here, for the first time, we successfully developed a novel, versatile, and controllable strategy for preparing PNDs via an anodized aluminum oxide (AAO) template-assisted method. With this approach, we prepared PNDs of an anticancer drug (VM-26) with precisely controlled sizes reaching the sub-20 nm range. This template-assisted approach has much higher feasibility for mass production comparing to the conventional reprecipitation method and is beneficial for future clinical translation. The present method is further demonstrated to be easily applicable for a wide range of hydrophobic biomolecules without the need of custom molecular modifications and can be extended for preparing all-in-one nanostructures with different functional agents.

In vivo tumor-targeted dual-modal fluorescence/CT imaging using a nanoprobe co-loaded with an aggregation-induced emission dye and gold nanoparticles.

As an intensely studied computed tomography (CT) contrast agent, gold nanoparticle has been suggested to be combined with fluorescence imaging modality to offset the low sensitivity of CT. However, the strong quenching of gold nanoparticle on fluorescent dyes requires complicated design and shielding to overcome. Herein, we report a unique nanoprobe (M-NPAPF-Au) co-loading an aggregation-induced emission (AIE) red dye and gold nanoparticles into DSPE-PEG(2000) micelles for dual-modal fluorescence/CT imaging. The nanoprobe was prepared based on a facile method of "one-pot ultrasonic emulsification". Surprisingly, in the micelles system, fluorescence dye (NPAPF) efficiently overcame the strong fluorescence quenching of shielding-free gold nanoparticles and retained the crucial AIE feature. In vivo studies demonstrated the nanoprobe had superior tumor-targeting ability, excellent fluorescence and CT imaging effects. The totality of present studies clearly indicates the significant potential application of M-NPAPF-Au as a dual-modal non-invasive fluorescence/X-ray CT nanoprobe for in vivo tumor-targeted imaging and diagnosis.

Aggregation-induced near-infrared absorption of squaraine dye in an albumin nanocomplex for photoacoustic tomography in vivo.

Photoacoustic tomography (PAT) is a newly emerging noninvasive imaging modality that could be further enhanced using near-infrared (NIR)-absorbing materials as contrast agents. To date, the most extensively studied photoacoustic imaging agents are inorganic nanomaterials because organic materials with NIR-absorption capabilities are limited. In this study, a NIR-absorbing nanocomplex composed of a squaraine dye (SQ) and albumin was prepared based on the aggregation-induced NIR absorption of SQ. Through aggregation, the absorption spectrum of SQ was widened from the visible-light region to the NIR region, which facilitated photoacoustic signal generation in the tissue-transparent NIR optical window (700-900 nm). Blood analysis and histology measurements revealed that the nanocomplex can be used for PAT applications in vivo without obvious toxicity to living mice.

Nanostructural systems developed with positive charge generation to drug delivery.

The surface charge of a nanostructure plays a critical role in modulating blood circulation time, nanostructure-cell interaction, and intracellular events. It is unfavorable to have positive charges on the nanostructure surface before arriving at the disease site because positively charged nanostructures interact strongly with blood components, resulting in rapid clearance from the blood, and suboptimal targeted accumulation at the tumor site. Once at the tumor site, however, the positive charge on the nanostructure surface accelerates uptake by tumor cells and promotes the release of payloads from the lysosomes to the cytosol or nucleus inside cells. Thus, the ideal nanocarrier systems for drug delivery would maintain a neutral or negatively charged surface during blood circulation but would then generate a positive surface charge after accumulation at the tumor site or inside the cancer cells. This Progress Report focuses on the design and application of various neutral or negatively charged nanostructures that can generate a positive charge in response to the tumor microenvironment or an external stimulus.

Non-blinking, highly luminescent, pH- and heavy-metal-ion-stable organic nanodots for bio-imaging.

Heavy-metal-free, organic-small-molecule-based fluorescent nanodots (Sdots, 31-60 nm in diameter) based on 2,7-di(4-(diphenylamino)phenyl-2,1,3-benzothiadiazol-7-yl)-9,9'-spirobifluorene (Spiro-BTA) were prepared through a simple solution process. The Sdots show not only non-blinking and high-brightness fluorescence but also stability in various pH conditions and heavy metal ion solutions. More importantly, Spiro-BTA Sdots demonstrate obviously much higher brightness with very high signal-to-background ratio in Hela cell, compared with common CdSe/ZnS QDs. In addition, they exhibit large Stokes shifts, broad absorption spectra, and low toxicity to living cells which enable their applications as good fluorescence probes for bio-imaging. Further application of Sdots for folate receptor-mediated live-cell endocytosis was demonstrated by non-convalent modification with folic acid linked multidentate ligands.