Deep Red Light Driven Hydrogen Evolution by Heterojunction Polymer Dots for Diabetic Wound Healing.

We describe small heterojunction polymer dots (Pdots) with deep-red light catalyzed H2 generation for diabetic skin wound healing. The Pdots with donor/acceptor heterojunctions showed remarkably enhanced photocatalytic activity as compared to the donor or acceptor nanoparticles alone. We encapsulate the Pdots and ascorbic acid into liposomes to form Lipo-Pdots nanoreactors, which selectively scavenge ⋅OH radicals in live cells and tissues under 650 nm light illumination. The antioxidant capacity of the heterojunction Pdots is ~10 times higher than that of the single-component Pdots described previously. Under a total light dose of 360 J/cm2, the Lipo-Pdots nanoreactors effectively scavenged ⋅OH radicals and suppressed the expression of pro-inflammatory cytokines in skin tissues, thereby accelerating the healing of skin wounds in diabetic mice. This study provides a feasible solution for safe and effective treatment of diabetic foot ulcers.

Site-Specific Modification of Single Domain Antibodies by Enzyme-Immobilized Magnetic Beads.

Nanobodies as imaging agents and drug conjugates have shown great potential for cancer diagnostics and therapeutics. However, site-specific modification of a nanobody with microbial transglutaminase (mTGase) encounters problems in protein separation and purification. Here, we describe a facile yet reliable strategy of immobilizing mTGase onto magnetic beads for site-specific nanobody modification. The mTGase immobilized on magnetic beads (MB-mTGase) exhibits catalytic activity nearly equivalent to that of the free mTGase, with good reusability and universality. Magnetic separation simplifies the protein purification step and reduces the loss of nanobody bioconjugates more effectively than size exclusion chromatography. Using MB-mTGase, we demonstrate site-specific conjugation of nanobodies with fluorescent dyes and polyethylene glycol molecules, enabling targeted immunofluorescence imaging and improved circulation dynamics and tumor accumulation in vivo. The combined advantages of MB-mTGase method, including high conjugation efficiency, quick purification, less protein loss, and recycling use, are promising for site-specific nanobody functionalization and biomedical applications.

Long-Term In Vivo Glucose Monitoring by Polymer-Dot Transducer in an Injectable Hydrogel Implant.

Optical sensors have attracted a great deal of interest for glucose detection. However, their practical applications for continuous glucose monitoring are still constrained by operational reliability in subcutaneous tissues. Here, we show an implantable hydrogel platform embedded with luminescent polymer dots (Pdots) for sensitive and long-term glucose monitoring. We use Pdot transducer in a polyacrylamide hydrogel matrix to construct an implantable platform. The hydrogel-Pdot transducer showed bright luminescence with ratiometric response to glucose changes. The in vitro and in vivo sensitivities of the hydrogel implant were enhanced by varying the enzyme concentration and injection volume. After implantation, the hydrogel with Pdot transducer remained at the implanted site without migration for 1 month and can be removed from the subcutaneous tissue for further analysis. Our results indicate that the hydrogel-Pdot platform maintains the intrinsic sensing property with excellent stability during 1 month implantation, while fibrous capsule formation on the implant in some cases needs to be solved for long-term continuous glucose monitoring.

NIR-II Fluorescence Imaging Reveals Bone Marrow Retention of Small Polymer Nanoparticles.

The concept that systemically administered nanoparticles are highly accumulated into the liver, spleen and kidney is a central paradigm in the field of nanomedicine. Here, we report that bone is an important organ for retention of small polymer nanoparticles using in vivo fluorescence imaging in the second near-infrared (NIR-II) window. We prepared different sized polymer nanoparticles with both visible and NIR-II fluorescence. NIR-II imaging reveals that the behavior of nanoparticle distribution in bone was largely dependent on the particle size. Small polymer nanoparticles of ∼15 nm diameter showed fast accumulation and long-term retention in bone, while the nanoparticles larger than ∼25 nm were dominantly distributed in liver. Confocal microscopy of bone sections indicated that the nanoparticles were largely distributed in the endothelial cells of sinusoidal vessels in bone marrow. The study provides promising opportunities for bone imaging and treatment of bone-related disease.

Light-Harvesting Fluorescent Spherical Nucleic Acids Self-Assembled from a DNA-Grafted Conjugated Polymer for Amplified Detection of Nucleic Acids.

The ultralow concentration of nucleic acids in complex biological samples requires fluorescence probes with high specificity and sensitivity. Herein, a new kind of spherical nucleic acids (SNAs) is developed by using fluorescent π-conjugated polymers (FCPs) as a light-harvesting antenna to enhance the signal transduction of nucleic acid detection. Specifically, amphiphilic DNA-grafted FCPs are synthesized and self-assemble into FCP-SNA structures. Tuning the hydrophobicity of the graft copolymer can adjust the size and light-harvesting capability of the FCP-SNAs. We observe that more efficient signal amplification occurs in larger FCP-SNAs, as more chromophores are involved, and the energy transfer can go beyond the Förster radius. Accordingly, the optimized FCP-SNA shows an antenna effect of up to 37-fold signal amplification and the limit of detection down to 1.7 pM in microRNA detection. Consequently, the FCP-SNA is applied to amplified in situ nucleic acid detecting and imaging at the single-cell level.

Semiconducting Polymer Dots with Dual-Enhanced NIR-IIa Fluorescence for Through-Skull Mouse-Brain Imaging.

Fluorescence probes in the NIR-IIa region show drastically improved imaging owing to the reduced photon scattering and autofluorescence in biological tissues. Now, NIR-IIa polymer dots (Pdots) are developed with a dual fluorescence enhancement mechanism. First, the aggregation induced emission of phenothiazine was used to reduce the nonradiative decay pathways of the polymers in condensed states. Second, fluorescence quenching was minimized by different levels of steric hindrance to further boost the fluorescence. The resulting Pdots displayed a fluorescence QY of ca. 1.7 % in aqueous solution, suggesting an enhancement of ca. 21 times in comparison with the original polymer in tetrahydrofuran (THF) solution. Small-animal imaging by using the NIR-IIa Pdots exhibited a remarkable improvement in penetration depth and signal to background ratio, as confirmed by through-skull and through-scalp fluorescent imaging of the cerebral vasculature of live mice.

Cooperative Blinking from Dye Ensemble Activated by Energy Transfer for Super-resolution Cellular Imaging.

Photoblinking is a fundamental process that occurs exclusively in single fluorophores such as organic dyes, fluorescent proteins, and quantum dots. Here, we describe a strategy to achieve pronounced, high on/off ratio, and cooperative blinking in donor-acceptor multifluorophore systems. An ensemble of dye molecules doped in semiconducting polymer dots (Pdots) exhibit robust photoblinking, while the pristine Pdots and the dye in optically inert polymer matrices fluoresce continuously. Energy transfer from Pdots to dye acceptors produces photoblinking via a cooperative process, in which the bright state originates from the dye ensemble and the dark state is due to quenching of semiconducting polymer by hole polarons. Using the blinking Pdots in subcellular structures labeling, we demonstrated approximately 3.6-fold enhancement of imaging resolution in high-order super-resolution optical fluctuation nanoscopy as compared to conventional microscopy. Our findings not only demonstrate the exciting possibility of transforming a nonquantized ensemble into a single-emitter-like optical source but also provide an effective approach to generate superior photoblinking fluorescent probes for super-resolution imaging applications.

Compact Conjugated Polymer Dots with Covalently Incorporated Metalloporphyrins for Hypoxia Bioimaging.

Hypoxia is closely related to multiple diseases, especially in tumors, which increases the aggressiveness and drug resistance of cancer cells. Precise hypoxia imaging is of great significance for cancer diagnosis and the evaluation of therapeutic effects. A kind of hydrophobic polymer (i.e., PFPtTFPP) as an imaging probe for hypoxia with fluorene as an energy donor and an oxygen-sensitive PtII porphyrin as an energy acceptor was developed. Compact polymer dots (Pdots) with a small size were prepared by nanoprecipitation. The PFPtTFPP Pdots showed excellent hypoxia sensing in solution with high sensitivity and full reversibility. The emission intensity, quantum yields, lifetime, and single-particle brightness significantly increased under hypoxia conditions. Remarkably, hypoxia imaging in vitro and in vivo was realized, and a clear increase in brightness was observed under hypoxia conditions and in the tumor area. Excellent hypoxia imaging ability is beneficial to potential applications in cancer diagnosis.

Semiconducting Polymer Nanocavities: Porogenic Synthesis, Tunable Host-Guest Interactions, and Enhanced Drug/siRNA Delivery.

Nanocavities composed of lipids and block polymers have demonstrated great potential in biomedical applications such as sensors, nanoreactors, and delivery vectors. However, it remains a great challenge to produce nanocavities from fluorescent semiconducting polymers owing to their hydrophobic rigid polymer backbones. Here, we describe a facile, yet general strategy that combines photocrosslinking with nanophase separation to fabricate multicolor, water-dispersible semiconducting polymer nanocavities (PNCs). A photocrosslinkable semiconducting polymer is blended with a porogen such as degradable macromolecule to form compact polymer dots (Pdots). After crosslinking the polymer and removing the porogen, this approach yields semiconducting polymer nanospheres with open cavities that are tunable in diameter. Both small molecules and macromolecules can be loaded in the nanocavities, where molecular size can be differentiated by the efficiency of the energy transfer from host polymer to guest molecules. An anticancer drug doxorubicin (Dox) is loaded into the nanocavities and the intracellular release is monitored in real time by the fluorescence signal. Finally, the efficient delivery of small interfering RNA (siRNA) to silence gene expression without affecting cell viability is demonstrated. The combined features of bright fluorescence, tunable cavity, and efficient drug/siRNA delivery makes these nanostructures promising for biomedical imaging and drug delivery.

Dual-Functional Ultrafiltration Membrane for Simultaneous Removal of Multiple Pollutants with High Performance.

Simultaneous removal of multiple pollutants from aqueous solution with less energy consumption is crucial in water purification. Here, a novel concept of dual-functional ultrafiltration (DFUF) membrane is demonstrated by entrapment of nanostructured adsorbents into the finger-like pores of ultrafiltration (UF) membrane rather than in the membrane matrix in previous reports of blend membranes, resulting in an exceptionally high active content and simultaneous removal of multiple pollutants from water due to the dual functions of rejection and adsorption. As a demonstration, hollow porous Zr(OH)x nanospheres (HPZNs) were immobilized in poly(ether sulfone) (PES) UF membranes through polydopamine coating with a high content of 68.9 wt %. The decontamination capacity of DFUF membranes toward multiple model pollutants (colloidal gold, polyethylene glycol (PEG), Pb(II)) was evaluated against a blend membrane. Compared to the blend membrane, the DFUF membranes showed 2.1-fold increase in the effective treatment volume for the treatment of Pb(II) contaminated water from 100 ppb to below 10 ppb (WHO drinking water standard). Simultaneously, the DFUF membranes effectively removed the colloidal gold and PEG below instrument detection limit, however the blend membrane only achieved 97.6% and 96.8% rejection for colloidal gold and PEG, respectively. Moreover, the DFUF membranes showed negligible leakage of nanoadsorbents during testing; and the membrane can be easily regenerated and reused. This study sheds new light on the design of high performance multifunction membranes for drinking water purification.

Incorporation of Porphyrin to π-Conjugated Backbone for Polymer-Dot-Sensitized Photodynamic Therapy.

The photosensitizers used in photodynamic therapy are mainly based on porphyrin derivatives. However, clinical applications encounter several limitations regarding photosensitizers such as their low absorption coefficients, poor water-solubility, and leaching from delivery carriers. Here, we describe covalent incorporation of porphyrin in conjugated polymer backbone for development of efficient polymer-dot photosensitizer. Spectroscopic characterizations revealed that the light-harvesting polymer dominantly transfer the excitation energy to the porphyrin unit, yielding efficient singlet oxygen generation for photodynamic therapy. The polymer dots (Pdots) also possess excellent stability that overcomes the photosensitizer leaching problem as encountered in other nanoparticle carriers. In vitro cytotoxicity and photodynamic efficacy of the Pdots were evaluated in MCF-7 cells by in vitro assay, indicating that the Pdots can efficiently damage cancer cells. In vivo photodynamic therapy by using the Pdots was further investigated with xenograft tumors in Balb/c nude mice, which show that the tumors were significantly inhibited or eradicated in certain cases. The high-yield singlet oxygen generation and excellent stability of porphyrin-incorporated Pdots are promising for photodynamic treatment of malignant tumors.

Near-Infrared II Semiconducting Polymer Dots: Chain Packing Modulation and High-Contrast Vascular Imaging in Deep Tissues.

Fluorescence imaging in the second near-infrared (NIR-II) window has attracted considerable interest in investigations of vascular structure and angiogenesis, providing valuable information for the precise diagnosis of early stage diseases. However, it remains challenging to image small blood vessels in deep tissues because of the strong photon scattering and low fluorescence brightness of the fluorophores. Here, we describe our combined efforts in both fluorescent probe design and image algorithm development for high-contrast vascular imaging in deep turbid tissues such as mouse and rat brains with intact skull. First, we use a polymer blending strategy to modulate the chain packing behavior of the large, rigid, NIR-II semiconducting polymers to produce compact and bright polymer dots (Pdots), a prerequisite for in vivo fluorescence imaging of small blood vessels. We further developed a robust Hessian matrix method to enhance the image contrast of vascular structures, particularly the small and weakly fluorescent vessels. The enhanced vascular images obtained in whole-body mouse imaging exhibit more than an order of magnitude improvement in the signal-to-background ratio (SBR) as compared to the original images. Taking advantage of the bright Pdots and Hessian matrix method, we finally performed through-skull NIR-II fluorescence imaging and obtained a high-contrast cerebral vasculature in both mouse and rat models bearing brain tumors. This study in Pdot probe development and imaging algorithm enhancement provides a promising approach for NIR-II fluorescence vascular imaging of deep turbid tissues.

Complexation modelling and oxidation mechanism of organic pollutants in cotton pulp black liquor during iron salt precipitation and electrochemical treatment.

Removal behavior of organic pollutants such as lignin in cotton pulp black liquor (CPBL) was investigated in precipitation followed by electrochemical oxidation (EO) using FeCl3, Fe2(SO4)3, FeCl2 and FeSO4 as precipitants, electrolyte and catalysts. Based on comparison of precipitation efficacy of iron salts, spectroscopic techniques, thermodynamic equilibrium calculations and molecular dynamics (MD) simulations were used to provide insight into the interaction between iron cations and lignin. The results showed that FeCl3 achieved the highest removal of chemical oxygen demand (COD, 76.05%), UV254 (69.21%) and lignin (78.28%). Iron cationic complexation with lignin was identified as the key mechanism in precipitation. Fe3+ was more active in binding to organic ligands mainly due to charge effect compared to Fe2+. The strong Fe-sulphate coordination affected the complexation with lignin. MD simulations showed the formation of inner sphere complexes of iron cations with deprotonated carboxyl and hydroxyl groups via bidentate and monodentate coordination. The removal efficiency of electrochemical oxidation (EO) as a post-treatment of the precipitation was dependent on iron salts. Removals of COD, UV254 and color can achieve 98.88%, 98.9% and 99.97% by FeCl3 precipitation and EO processes. The effluent reached the primary discharge standard specified in Integrated Wastewater Discharge Standard of China (GB8978-1996). FeCl3 demonstrated significant advantages in the removal of organic pollutants from cotton pulp black liquor in the combined process of precipitation and electrochemical treatment and may have practical application potential.

Surface Plasmon-Enhanced NIR-II Fluorescence in a Multilayer Nanoprobe for Through-Skull Mouse Brain Imaging.

Fluorescence imaging in the second near-infrared window (NIR-II, 1000-1700 nm) holds great potential for the accurate visualization of deeply located biological structures in vivo. However, the weak fluorescence of current NIR-II fluorophores remains a long-standing challenge for the ever-growing imaging demand. Here, we describe a surface plasmon-enhanced NIR-II fluorescence strategy by incorporating silica-coated gold nanorods (GNRs) and polymer dots (Pdots) into a multilayer nanostructure. Precise manipulation of the silica spacing layer thickness signifies an optimum distance of 8.6 nm, where an enhancement factor of up to 6.4 is achieved in the NIR-II imaging window. The surface plasmon enhancement approach is successfully extended to several types of Pdots fluorophores with NIR-II emission. We finally perform outer-layer encapsulation and PEGylation for the multilayer probes and demonstrate surface plasmon-enhanced NIR-II fluorescence for mouse brain imaging through the skull, which exhibits a refined signal-to-background ratio and penetration depth as compared to the clinically approved ICG dye.

Light-Controlled Precise Delivery of NIR-Responsive Semiconducting Polymer Nanoparticles with Promoted Vascular Permeability.

The endothelial barrier plays an essential role in health and disease by protecting organs from toxins while allowing nutrients to access the circulation. However, it is the major obstacle that limits the delivery of therapeutic drugs to the diseased tissue. Here, it is reported for the first time that near-infrared (NIR) laser pulses can transiently promote the delivery of semiconducting polymer nanoparticles passing the vascular barrier via photoacoustic-effect-induced accumulation, only by the aid of pulse laser irradiation. This strategy enables selective and substantial accumulation of the NIR-absorbing nanoparticles inside specific tissues, implying the discovery of an unprecedented approach for light-controlled nanoparticle delivery. Especially, the nanoparticle delivery in solid tumors by 10-min laser scanning is approximately six times higher than that of the enhanced permeability and retention (EPR) effect in 24 h under current experimental conditions. Further results confirm that this strategy facilitates substantial accumulation of nanoparticles in the mouse brain with intact skull. This approach thus opens a new door for tissue-specific delivery of nanomaterials with an unprecedented level of efficiency and precision.

A biodegradable nano-photosensitizer with photoactivatable singlet oxygen generation for synergistic phototherapy.

Photodynamic therapy (PDT) is a promising method for cancer therapy and also may initiate unexpected damages to normal cells and tissues. Herein, we develop a near-infrared (NIR) light-activatable nanophotosensitizer, which shows negligible phototoxicity before photoactivation to improve the specificity of PDT. The nanophotosensitizer is prepared by indocyanine green carboxylic (ICG), Chlorin e6 (Ce6), and biodegradable poly (lactic acid) (PLA) and poly (lactic-co-glycolic acid) (PLGA), and all these materials have been approved by the Food and Drug Administration. Initially the phototoxicity of Ce6 is effectively inhibited by ICG through fluorescence resonance energy transfer (FRET). Upon 808 nm laser activation, ICG generate hyperthermia for photothermal therapy (PTT) and simultaneously is degraded due to the inherently poor photostability. The FRET is disrupted and followed by the recovery of phototoxicity of Ce6 for PDT. We investigated the photoactivation and the resulting phototherapy by cellular assays and mouse models, which indicate a superior synergistic treatment effect and selective PDT activated by near-infrared 808 nm light. This study presents a promising strategy for activatable and synergistic phototherapy with minimal damage to normal tissues.

Brightness Enhancement of Near-Infrared Semiconducting Polymer Dots for in Vivo Whole-Body Cell Tracking in Deep Organs.

In vivo visualization of cell migration and engraftment in small animals provide crucial information in biomedical studies. Semiconducting polymer dots (Pdots) are emerging as superior probes for biological imaging. However, in vivo whole-body fluorescence imaging is largely constrained by the limited brightness of Pdots in near-infrared (NIR) region. Here, we describe the brightness enhancement of NIR fluorescent Pdots for in vivo whole-body cell tracking in deep organs. We first synthesize semiconducting polymers with strong absorption in orange and far-red regions. By molecular doping, the weak broad-band fluorescence of the Pdots was significantly narrowed and enhanced by 1 order of magnitude enhancement, yielding bright narrow-band NIR emission with a quantum yield of ∼0.21. Under an excitation of far-red light (676 nm), a trace amount of Pdots (∼2 µg) in the stomach can be clearly detected in whole-body fluorescence imaging of live mice. The Pdots coated with a cell-penetrating peptide are able to brightly label cancer cells with minimal cytotoxicity. In vivo cell tracking in live mice indicated that the entrapment and migration of the tail-vein-administered cells (∼400 000) were clearly visualized in real time. These Pdots with deep-red excitation and bright NIR emission are promising for in vivo whole-body fluorescence imaging.

A synergetic analysis method for antifouling behavior investigation on PES ultrafiltration membrane with self-assembled TiO2 nanoparticles.

Fouling of ultrafiltration (UF) membranes is a major impediment for their use in drinking water production. Mixed matrix membranes (MMMs) may have great opportunities in dealing with this challenge due to their hierarchical structures and multiple functionalities. In this study, a synergetic analysis method based on intermolecular adhesion force measurement and fouling process simulation was applied to investigate the fouling mechanism of polyethersulfone (PES) UF membranes containing in situ self-assembled TiO2 nanoparticles (NPs). The fouling resistance behavior and antifouling mechanism of the newly developed composite membranes were investigated with sodium alginate (SA), bovine serum albumin (BSA) and humic acid (HA) as model organic foulants. An improved antifouling effect was conspicuously observed for the composite membranes, expressed by a lower flux decline and significantly better cleaning efficiency. A strong correlation between the self-assembled structure of TiO2 NPs and the antifouling behavior of the composite membrane was observed. A lower magnitude and a narrower distribution of adhesion forces for the composite membrane suggest the effective suppression of foulants adsorption on the clean or fouled membrane. The simulation analysis indicates that the main fouling mechanism was standard blocking and cake filtration, further confirming the superiority of the NPs self-assembled structure in mitigating membrane fouling. This dual analysis method may provide a promising technological support for the application of modified UF membranes with self-assembled NPs in drinking water production.