Xin Ting Zheng

Glowing Graphene Quantum Dots and Carbon Dots: Properties, Syntheses, and Biological Applications

The emerging graphene quantum dots (GQDs) and carbon dots (C-dots) have gained tremendous attention for their enormous potentials for biomedical applications, owing to their unique and tunable photoluminescence properties, exceptional physicochemical properties, high photostability, biocompatibility, and small size. This article aims to update the latest results in this rapidly evolving field and to provide critical insights to inspire more exciting developments. We comparatively review the properties and synthesis methods of these carbon nanodots and place emphasis on their biological (both fundamental and theranostic) applications.

One-step and high yield simultaneous preparation of single- and multi-layer graphene quantum dots from CX-72 carbon black

A facile, low cost and high yield method has been developed to prepare single- and multi-layer graphene quantum dots (GQDs) from XC-72 carbon black by chemical oxidation. The single-layer GQDs are demonstrated to be excellent probes for cellular imaging, while the multi-layer GQDs may offer great potential applications in optoelectronic devices.

Graphene quantum dots as universal fluorophores and their use in revealing regulated trafficking of insulin receptors in adipocytes

Graphene quantum dots (GQDs) hold great promise as a new class of fluorophores for bioimaging, owing to their remarkable physicochemical properties including tunable photoluminescence, excellent photostability, and biocompatibility. Despite their highly anticipated potentials, GQDs have yet to be used to specifically label and track molecular targets involved in dynamic cellular processes in live cells. Here, we demonstrate that GQDs can serve as universal fluorophores for bioimaging because they can be readily conjugated with a wide range of biomolecules while preserving their functionalities. As a proof-of-concept demonstration, insulin-conjugated GQDs have been synthesized and utilized for specific labeling and dynamic tracking of insulin receptors in 3T3-L1 adipocytes. Our experiments reveal, for the first time, that the internalization and recycling of insulin receptors in adipocytes are oppositely regulated by apelin and TNFα, which may contribute to the regulations of these two cytokines in insulin sensitivity.

RGD-Peptide Functionalized Graphene Biomimetic Live-Cell Sensor for Real-Time Detection of Nitric Oxide Molecules

It is always challenging to construct a smart functional nanostructure with specific physicochemical properties to real time detect biointeresting molecules released from live-cells. We report here a new approach to build a free-standing biomimetic sensor by covalently bonding RGD-peptide on the surface of pyrenebutyric acid functionalized graphene film. The resulted graphene biofilm sensor comprises a well-packed layered nanostructure, in which the RGD-peptide component provides desired biomimetic properties for superior human cell attachment and growth on the film surface to allow real-time detection of nitric oxide, an important signal yet short-life molecule released from the attached human endothelial cells under drug stimulations. The film sensor exhibits good flexibility and stability by retaining its original response after 45 bending/relaxing cycles and high reproducibility from its almost unchanged current responses after 15 repeated measurements, while possessing high sensitivity, good selectivity against interferences often existing in biological systems, and demonstrating real time quantitative detection capability toward nitric oxide molecule released from living cells. This study not only demonstrates a facial approach to fabricate a smart nanostructured graphene-based functional biofilm, but also provides a powerful and reliable platform to the real-time study of biointeresting molecules released from living cells, thus rendering potential broad applications in neuroscience, screening drug therapy effect, and live-cell assays.

In situ molecular detection of ischemic cells by enhanced protein direct electron transfer on a unique horseradish peroxidase–Au nanoparticles–polyaniline nanowires biofilm

To overcome shortcomings of the ex situ approaches, in situ detection using H(2)O(2) molecules to diagnose ischemia through enhanced protein direct electron transfer on a unique horseradish peroxidase-Au nanoparticles-polyaniline nanowires biofilm is demonstrated and it is discovered that the extracellular H(2)O(2) molecule released per ischemic cell is 2.7-times of that of a normal cell.

Restoring basal planes of graphene oxides for highly efficient loading and delivery of β-lapachone.

An efficient and biocompatible drug nanocarrier is essential for nanomedicines to realize their full therapeutic potential. Here, we investigate the loading of a selective and potent anticancer drug, β-lapachone (β-lap), on a magnetite nanoparticle-decorated reduced graphene oxide (Fe(3)O(4)/rGO) and the in vitro anticancer efficacy of β-lap loaded Fe(3)O(4)/rGO. Reduced graphene oxide (rGO) with magnetic functionality was prepared via electrostatic interaction between positively charged magnetite (Fe(3)O(4)) nanoparticles and negatively charged GO, followed by hydrazine reduction of GO to rGO. The prepared Fe(3)O(4)/rGO shows that Fe(3)O(4) makes the Fe(3)O(4)/rGO hybrid magnetically separable for easy handling during drug loading and release and the Fe(3)O(4)/rGO hybrid exhibits significantly higher loading capacity than that of Fe(3)O(4)/GO, suggesting that restoration of the graphene basal plane upon reduction of GO enhances the interaction between β-lap and rGO. Cellular uptake studies using fluorescently labeled Fe(3)O(4)/rGO verifies successful internalization of Fe(3)O(4)/rGO into the cytoplasm while rGO without hybridized Fe(3)O(4) has poor uptake performance. Furthermore, β-lap loaded Fe(3)O(4)/rGO shows remarkably high cytotoxicity toward MCF-7 breast cancer cells while the blank Fe(3)O(4)/rGO produces no cytotoxic effects. The cytotoxicity results suggest that Fe(3)O(4)/rGO is an efficient drug carrier for anticancer treatments. The fine-tuning of the chemical structures of graphene oxides by reduction chemistry may provide a universal route for controlled loading and release of drugs or biomolecules to construct advanced delivery vehicles.

Optical Detection of Single Cell Lactate Release for Cancer Metabolic Analysis

Sensitive detection of extracellular lactate concentrations at a single cell level is of importance for studying the metabolic alterations in tumor progression. A unique nanoscale optical fiber lactate sensor was developed to monitor the extracellular lactate concentrations of cancer cells by immobilizing its nanotip with lactate dehydrogenases, which could catalyze lactate conversion to generate NADH for sensitive fluorescence detection. The results demonstrate that the fabricated nanosensor can successfully detect the extracellular lactate concentrations for single HeLa, MCF-7, and human fetal osteoblast (hFOB) cells, showing that the cancer cells have distinctly higher extracellular lactate concentrations than normal cells as that predicted by Warburg effect. The nanosensor was also employed to investigate the effect of a monocarboxylate transporter inhibitor on the lactate efflux from cancer cells. Different lactate efflux inhibition profiles were obtained for HeLa and MCF-7 cell lines. This work demonstrates that the nanosensor has potential for evaluating the effect of metabolic agents on cancer metabolism and survival.

Single living cell detection of telomerase over-expression for cancer detection by an optical fiber nanobiosensor.

An optical fiber nanobiosensor was constructed to successfully detect a general cancer biomarker, telomerase at single cell level with its nanoscale tip. The nanotip immobilized with a specific antibody was inserted into a MCF-7 breast cancer cell nucleus to capture telomerases directly, after which an in vitro enzymatic sandwich immunoassay was performed to achieve sensitive single living cell detection. The nanotip inserted into MCF-7 cell nucleus provides significantly higher average (F-F(0))/F(0) ratio than that of human mesenchymal stem cell (hMSC) nucleus, demonstrating the successful detection of the telomerase over-expression in cancer cells as compared to normal cells. The detection in the cytoplasm shows much smaller average ratio than that in the nucleus of MCF-7 cells while clearly verifies the nuclear localization of telomerase. The successful detection of telomerase over-expression in a single living cell for the first time may provide a potential method for cancer detection, and also demonstrate a universal approach that can be used to detect other low expression proteins in a single living cell.

Silica-based complex nanorattles as multifunctional carrier for anticancer drug

In this work, we demonstrate a new route to generate silica-based multifunctional complex nanorattles. By using SnO2 hollow nanospheres as the starting template, we are able to build a new type of rattle-in-ball hollow structure with multi-level interior architecture, where the core of a nanorattle is itself another smaller rattle. Moreover, additional functionality can be introduced by forming Au nanoparticles in the interstitial space. Such unique hollow nanostructures are believed to be very useful as nanoreactors for selective catalysis. Magnetic functionality can also be incorporated by using an α-Fe2O3 nanospindle as the starting template followed by in situreduction to Fe3O4. This type of ellipsoidal nanorattle is feasible for drug delivery as it is shown to be highly biocompatible and non-cytotoxic. The cell viability assay proves that the sample is an efficient drug delivery vehicle exhibiting similar anticancer efficacy against MCF-7 carcinoma cells as compared to the free DOX. By constructing these two types of complex nanorattles as examples, we demonstrate new possible routes to generate versatile hollow nanostructures with distinct architectures and chemical compositions, thus widening their application potential.

Bifunctional electro-optical nanoprobe to real-time detect local biochemical processes in single cells

A bifunctional electro-optical nanoprobe with integrated nanoring electrode and optical nanotip was fabricated and investigated to simultaneously detect both electrical and optical signals in real-time with high spatial resolution. Concurrent measurements of the oxidant generation and the intracellular antioxidant levels in single cells correlate the stronger oxidant generation with an altered initial antioxidant response in the breast cancer cells in comparison to the normal ones suggesting that the cell malignancy is associated with the strength of oxidative stress, and the higher antioxidant level may be the cause of the drug resistance. While the optical detection indicates the fluctuation of the intracellular redox homeostasis, the chronoamperometric signals allow quantitative real-time detection of the H₂O₂ release and decay. Furthermore, the nanoscale probe enables localized simultaneous detections thus discovering that activated enzymes responsible for the oxidative stress target at specific membrane regions. This method promises applications in study of the dynamics of important physiological processes, and provides the opportunity to unravel the interplay of various signaling pathways.