Carbon Dots with Absorption Red-Shifting for Two-Photon Fluorescence Imaging of Tumor Tissue pH and Synergistic Phototherapy.

Phototherapy exhibits significant potential as a novel tumor treatment method, and the development of highly active photosensitizers and photothermal agents has drawn considerable attention. In this work, S and N atom co-doped carbon dots (S,N-CDs) with an absorption redshift effect were prepared by hydrothermal synthesis with lysine, o-phenylenediamine, and sulfuric acid as raw materials. The near-infrared (NIR) absorption features of the S,N-CDs resulted in two-photon (TP) emission, which has been used in TP fluorescence imaging of lysosomes and tumor tissue pH and real-time monitoring of apoptosis during tumor phototherapy, respectively. The obtained heteroatom co-doped CDs can be used not only as an NIR imaging probe but also as an effective photodynamic therapy/photothermal therapy (PDT/PTT) therapeutic agent. The efficiencies of different heteroatom-doped CDs in tumor treatment were compared. It was found that the S,N-CDs showed higher therapeutic efficiency than N-doped CDs, the efficiency of producing 1O2 was 27%, and the photothermal conversion efficiency reached 34.4%. The study provides new insight into the synthesis of carbon-based nanodrugs for synergistic phototherapy and accurate diagnosis of tumors.

A tumor microenvironment-induced absorption red-shifted polymer nanoparticle for simultaneously activated photoacoustic imaging and photothermal therapy.

Tumor microenvironment-responsive therapy has enormous application potential in the diagnosis and treatment of cancer. The glutathione (GSH) level has been shown to be significantly increased in tumor tissues. Thus, GSH can be used as an effective endogenous molecule for diagnosis and tumor microenvironment-activated therapy. In this study, we prepared a tumor microenvironment-induced, absorption spectrum red-shifted, iron-copper co-doped polyaniline nanoparticle (Fe-Cu@PANI). The Cu(II) in this nanoparticle can undergo a redox reaction with GSH in tumors. The redox reaction induces a red shift in the absorption spectrum of the Fe-Cu@PANI nanoparticles from the visible to the near-infrared region accompanying with the etching of this nanoparticle, which simultaneously activates tumor photoacoustic imaging and photothermal therapy, thereby improving the accuracy of in vivo tumor imaging and the efficiency of photothermal therapy. The nanoparticle prepared in this study has broad application prospects in the diagnosis and treatment of cancer.

Accelerating the peroxidase-like activity of MoSe2 nanosheets at physiological pH by dextran modification.

Surface modification of MoSe2via dextran during ultrasound exfoliation is demonstrated to be an efficient and easy strategy to accelerate the peroxidase-like catalytic activity of MoSe2 nanosheets at neutral pH. The enhancement of catalytic activity is owing to the rich negative charges of dextran on the dextran-modified MoSe2 nanosheets.

Dual functionalized natural biomass carbon dots from lychee exocarp for cancer cell targetable near-infrared fluorescence imaging and photodynamic therapy.

Photodynamic therapy (PDT) is a non-invasive phototherapy that has gained significant attention for cancer therapy. However, image-guided PDT still remains a considerable challenge. Herein, we developed a targeted, near-infrared (NIR) fluorescence imaging nanoprobe for cancer cells by preparing natural biomass carbon dots (NBCDs) from lychee exocarp, and loading transferrin and a photosensitizer on the NBCD surfaces for image-guided PDT of cancer cells and mouse tumors. Because the surfaces of cancer cells exhibit more transferrin receptors, the proposed NIR fluorescent nanoprobe can better penetrate cancer cells for cancer cell targetable fluorescence imaging. Thus, the dual-function nanoprobe made from natural biomass can be used as a specific agent for NIR fluorescence imaging and PDT. More importantly, the functional nanoprobe prepared from natural biomass emits NIR fluorescence, shows very low biological toxicity, and can minimize side effects on normal cells. After directly injecting the nanoprobes into tumor tissues, the photosensitizers on the surface of the NBCDs can produce singlet oxygen (1O2) through photodynamic reactions when irradiated with 650 nm light to kill cancer cells, thus inhibiting tumor growth in PDT-treated mice. Therefore, the functional fluorescent nanoprobe made from natural biomass has been employed as a PDT agent, and holds great promise in image-guided tumor PDT.

Self-assembled nanomaterials for synergistic antitumour therapy.

Self-assembly is a bottom-up strategy that can be used to construct a wide range of nanostructures. This review discusses the applications of self-assembled nanomaterials in the field of antitumour therapy. A variety of materials have been developed, such as nanomaterials self-assembled from polymers, biomacromolecules, proteins, peptides, inorganic materials, nucleic acids, and organic molecules, among others, and their applications include phototherapy, chemotherapy, gene therapy, imaging, immunotherapy and other fields. These therapeutic methods can combine with each other as synergistic therapy. Here, we review the progress of self-assembled anticancer nanomaterials applied to target tumours, decrease drug resistance, cross the blood-brain barrier to treat metastases, and perform other tasks. Additional challenges to be overcome in this field are also discussed.

Nitrogen and Phosphorus Co-Doped Carbon Nanodots as a Novel Fluorescent Probe for Highly Sensitive Detection of Fe(3+) in Human Serum and Living Cells.

Chemical doping with heteroatoms can effectively modulate physicochemical and photochemical properties of carbon dots (CDs). However, the development of multi heteroatoms codoped carbon nanodots is still in its early stage. In this work, a facile hydrothermal synthesis strategy was applied to synthesize multi heteroatoms (nitrogen and phosphorus) codoped carbon nanodots (N,P-CDs) using glucose as carbon source, and ammonia, phosphoric acid as dopant, respectively. Compared with CDs, the multi heteroatoms doped CDs resulted in dramatic improvement in the electronic characteristics and surface chemical activities. Therefore, the N,P-CDs prepared as described above exhibited a strong blue emission and a sensitive response to Fe(3+). The N,P-CDs based fluorescent sensor was then applied to sensitively determine Fe(3+) with a detection limit of 1.8 nM. Notably, the prepared N,P-CDs possessed negligible cytotoxicity, excellent biocompatibility, and high photostability. It was also applied for label-free detection of Fe(3+) in complex biological samples and the fluorescence imaging of intracellular Fe(3+), which indicated its potential applications in clinical diagnosis and other biologically related study.

A carbon nanotubes based fluorescent aptasensor for highly sensitive detection of adenosine deaminase activity and inhibitor screening in natural extracts.

A carbon nanotubes (CNTs) based fluorescent aptasensor was developed for adenosine deaminase (ADA) activity detection and inhibitor screening by using adenosine (AD) as the substrate. This sensing system consists of CNTs, AD, split anti-AD aptamer fragment and dye-labeled aptamer fragment. In the absence of ADA, two aptamer fragments bind simultaneously with AD to form an AD-aptamer complex. This AD-aptamer complex cannot adsorb onto CNTs, and has high fluorescence intensity. When ADA is introduced into this system, ADA can convert AD into inosine, which has not affinity to the split anti-AD aptamer fragment. Thus, the split anti-AD aptamer fragments were adsorbed onto CNTs via strong π-π stacking interactions, resulting in the quenching of the fluorescence of the dye-labeled aptamer fragment. The proposed aptasensor can detect ADA activity from 0.005 to 0.2U/mL with a low detection limit of 0.002 U/mL. Moreover, it has been also demonstrated that this CNTs-based fluorescence aptasensor is suitable for ADA inhibitor screening from traditional Chinese medicine (TCM). Considering the superior sensitivity and specificity, the proposed CNTs-based fluorescent aptasensor can be expected to provide a simple, cost-effective and sensitive platform for the detection of ADA activity and screening of potential drugs.

Carbon nanotube signal amplification for ultrasensitive fluorescence polarization detection of DNA methyltransferase activity and inhibition.

A versatile sensing platform based on multiwalled carbon nanotube (MWCNT) signal amplification and fluorescence polarization (FP) is developed for the simple and ultrasensitive monitoring of DNA methyltransferase (MTase) activity and inhibition in homogeneous solution. This method uses a dye-labeled DNA probe that possess a doubled-stranded DNA (dsDNA) part for Mtase and its corresponding restriction endonuclease recognition, and a single-stranded DNA part for binding MWCNTs. In the absence of MTase, the dye-labeled DNA is cleaved by restriction endonuclease, and releases very short DNA carrying the dye that cannot bind to MWCNTs, which has relatively small FP value. However, in the presence of MTase, the specific recognition sequence in the dye-labeled DNA probe is methylated and not cleaved by restriction endonuclease. Thus, the dye-labeled methylated DNA product is adsorbed onto MWCNTs via strong π-π stacking interactions, which leads to a significant increase in the FP value due to the enlargement of the molecular volume of the dye-labeled methylated DNA/MWCNTs complex. This provides the basic of a quantitative measurement of MTase activity. By using the MWCNT signal amplification approach, the detection sensitivity can be significantly improved by two orders of magnitude over the previously reported methods. Moreover, this method also has high specificity and a wide dynamic range of over five orders of magnitude. Additionally, the suitability of this sensing platform for MTase inhibitor screening has also been demonstrated. This approach may serve as a general detection platform for sensitive assay of a variety of DNA MTases and screening potential drugs.

Screening α-glucosidase inhibitor from natural products by capillary electrophoresis with immobilised enzyme onto polymer monolith modified by gold nanoparticles.

A novel strategy for screening α-glucosidase inhibitors (AGIs) from natural products by capillary electrophoresis (CE) with an immobilised enzyme microreactor was developed. In this approach, gold nanoparticles (AuNPs) was first covalently attached to surface of the pores of the porous polymer capillary monolith via the formation of an Au-S bond, and α-glucosidase was then simply and stably immobilised onto AuNPs through the strong affinity of gold for amino groups of the enzyme. In order to profiling the activity of the immobilised α-glucosidase, the natural substrate was hydrolyzed by it and the yield of product was determined by CE. The amount of covalently attached α-glucosidase to the monolith was calculated to be about 30.0 µg/mg. The immobilised enzyme exhibited 80% activity after 25 runs, and only lost 7.6% of activity after 6 runs within 31 days. Screening of AGIs present in extracts of natural products by the proposed method was demonstrated.

A gold nanoparticle-mediated enzyme bioreactor for inhibitor screening by capillary electrophoresis.

A facile protocol to prepare highly effective and durable in-line enzyme bioreactors inside capillary electrophoresis (CE) columns was developed. To demonstrate the methodology, l-glutamic dehydrogenase (GLDH) was selected as the model enzyme. GLDH was first immobilized onto 38-nm-diameter gold nanoparticles (GNPs), and the functionalized GNPs were then assembled on the inner wall at the inlet end of the CE capillary treated with polyethyleneimine (PEI), producing an in-line GLDH bioreactor. Compared with a GLDH bioreactor prepared by immobilizing GLDH directly on PEI-treated capillary, the GNP-mediated bioreactor showed a higher enzymatic activity and a much better stability. The in-capillary enzyme bioreactor was proven to be very useful for screening of GLDH inhibitors deploying the GLDH-catalyzed α-ketoglutaric acid reaction. The screening assay was preliminarily validated by using a known GLDH inhibitor, namely perphenazine. A Z' factor value of 0.95 (n=10) was obtained, indicating that the screening results were highly reliable. Screening of GLDH inhibitors present in medicinal plant extracts by the proposed method was demonstrated. The inhibition percentages were found to be 53% for Radix scutellariae, 45% for Radix codonopsis, 37% for Radix paeoniae alba, and 0% for the other 22 extracts tested at a concentration of 0.6mg extract/ml.

Rapid screening of monoamine oxidase B inhibitors in natural extracts by capillary electrophoresis after enzymatic reaction at capillary inlet.

A facile capillary electrophoresis (CE) method was developed for the screening of monoamine oxidase B (MAO-B) inhibitors in natural extracts. In this method, the enzymatic reaction occurred at the capillary inlet during a predetermined waiting period, followed by the electrophoretic separation of the reaction compounds, and detected by their UV absorbance at 280 nm. Conditions for the separation of substrates, products and enzyme were optimized. The optimal buffer composition was 50 mM N-2-hydroxyethyl-piperazine-N'-2-ethane sulphonic acid (HEPES) solution containing 10 mM SDS (pH=7.4). Under the optimal condition, the baseline separation of substrates, products and enzyme was achieved within 2 min. The present method was used to determine MAO-B kinetic constants, K(i), K(m) and IC50 based on quantitative of the substrate peak area compared with the reference electropherogram obtained from without the inhibitor. A validation study shows good reproducibility for both migration time (RSD=1.8%) and peak area (RSD=3.9%). Finally, the screening of 16 natural extracts was performed, and 2 natural extracts from Fructus crataegi and Radix polygoni multiflori were identified to be positive for MAO-B inhibition.

Immobilized capillary adenosine deaminase microreactor for inhibitor screening in natural extracts by capillary electrophoresis.

A novel strategy for the preparation of in-column adenosine deaminase (ADA) microreactor and rapid screening of enzyme inhibitors in natural extracts was demonstrated. In this approach, ADA was encapsulated in anionic polyelectrolyte alginate that was immobilized on the surface of fused-silica capillary via ionic binding technique with cationic polyelectrolyte polyethylenimine (PEI). On-line enzyme inhibition study was performed by capillary electrophoresis (CE). The substrate and product were baselined separated within 75s. The enzyme activity was determined by the quantification of peak area of the product. Enzyme inhibition can be read out directly from the reduced peak area of the product in comparison with a reference electropherogram obtained in the absence of any inhibitor. The inhibition percentage was used to evaluate relative activity of ADA microreactor. A known ADA inhibitor, erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) was employed as a model compound for the validation of the inhibitor screening method, and the screening of ADA inhibitor in 19 traditional Chinese herbal medicines was performed.