Dual Nanoprobe Research Articles

Selective fluoride detection based on modulation of red emissive carbon dots fluorescence by zirconium-alizarin complex: Application to Nile River water and human saliva samples

Fluoride is an essential ion for dental health that is optimally supplemented through drinking water, toothpastes, and other oral hygiene products. This work reports a selective, simple, and inexpensive fluorescent sesnor for fluoride quantification. The sensor exploits the red fluorescence of carbon dots coupled with the colorimetric response of a zirconium-alizarin complex. The zirconium-alizarin exhibits an absorption band at 530 nm that overlaps the carbon dot excitation peak, resulting in inner filter effect-based quenching of the 612 nm red emission. However, in the presence of fluoride, binding disrupts the zirconium complex, releasing free alizarin with an absorption blue shifted to 420 nm. This reduces spectral overlap and restores carbon dot fluorescence. The carbon dots were synthesized by a hydrothermal method and exhibited excellent fluorescence quantum yield (16.3 %). Structural and optical characterizations were performed using TEM, XRD, XPS, FTIR, UV–vis, and fluorescence techniques. Under optimized conditions, the probe provided selective turn-on fluorescence detection of fluoride with a linear response from 1.0 to 130.0 μM and limit of detection (S/N = 3) of 0.32 μM. Practical utility was demonstrated by accurate quantification of fluoride in Nile River water and human saliva samples. This dual nanoprobe strategy offers a rapid and cost-effective platform for fluoride analysis in support of healthcare and environmental monitoring applications.

Selective fluorescence turn-on detection of combination cisplatin-etoposide chemotherapy based on N-CDs/GSH-CuNCs nanoprobe.

Cisplatin (CIS) and etoposide (ETP) combination therapy is highly effective for treating various cancers. However, the potential for pharmacokinetic interactions between these drugs necessitates selective sensing methods to quantitate both CIS and ETP levels in patient's plasma. This work develops a dual fluorescence probe strategy using glutathione-capped copper nanoclusters (GSH-CuNCs) and nitrogen-doped carbon dots (N-CDs) for the simultaneous analysis of CIS and ETP. The fluorescence signal of GSH-CuNCs at 615 nm increased linearly with CIS concentration while the N-CD emission at 480 nm remained unaffected. Conversely, the N-CD fluorescence was selectively enhanced by ETP with no interference with the CuNC fluorescence. Extensive materials characterization including UV-vis, fluorescence spectroscopy, XRD, and TEM confirmed the synthesis of the nanoprobes. The sensor showed high sensitivity with limits of detection of 6.95 ng mL-1 for CIS and 7.63 ng mL-1 for ETP along with excellent selectivity against potential interferences in rabbit plasma. Method feasibility was demonstrated with application to real rabbit plasma samples. The method was further applied to estimate the pharmacokinetic parameters of CIS before and after ETP coadministration. The dual nanoprobe sensing strategy enables rapid and selective quantitation of CIS and ETP levels to facilitate therapeutic drug monitoring and optimization of combination chemotherapy regimens.

Dual-labeled nanoparticles based on small extracellular vesicles for tumor detection

BackgroundSmall extracellular vesicles (sEVs) are emerging natural nanoplatforms in cancer diagnosis and therapy, through the incorporation of signal components or drugs in their structure. However, for their translation into the clinical field, there is still a lack of tools that enable a deeper understanding of their in vivo pharmacokinetics or their interactions with the cells of the tumor microenvironment. In this study, we have designed a dual-sEV probe based on radioactive and fluorescent labeling of goat milk sEVs.ResultsThe imaging nanoprobe was tested in vitro and in vivo in a model of glioblastoma. In vitro assessment of the uptake of the dual probe in different cell populations (RAW 264.7, U87, and HeLa) by optical and nuclear techniques (gamma counter, confocal imaging, and flow cytometry) revealed the highest uptake in inflammatory cells (RAW 264.7), followed by glioblastoma U87 cells. In vivo evaluation of the pharmacokinetic properties of nanoparticles confirmed a blood circulation time of ~ 8 h and primarily hepatobiliary elimination. The diagnostic capability of the dual nanoprobe was confirmed in vivo in a glioblastoma xenograft model, which showed intense in vivo uptake of the SEV-based probe in tumor tissue. Histological assessment by confocal imaging enabled quantification of tumor populations and confirmed uptake in tumor cells and tumor-associated macrophages, followed by cancer-associated fibroblasts and endothelial cells.ConclusionsWe have developed a chemical approach for dual radioactive and fluorescent labeling of sEVs. This methodology enables in vivo and in vitro study of these vesicles after exogenous administration. The dual nanoprobe would be a promising technology for cancer diagnosis and a powerful tool for studying the biological behavior of these nanosystems for use in drug delivery.Graphical

Technetium-99m and ICG-labeled HPG (hyperbranched polyglycerol) as a SPECT/FL dual imaging nanoprobe for imaging blood cells: in vitro investigation using myelogenous leukemia cells

Technetium-99m and ICG-labeled HPG (hyperbranched polyglycerol) as a SPECT/FL dual imaging nanoprobe for imaging blood cells: in vitro investigation using myelogenous leukemia cells

Asialoglycoprotein receptor targeted optical and magnetic resonance imaging and therapy of liver fibrosis using pullulan stabilized multi-functional iron oxide nanoprobe

Early diagnosis and therapy of liver fibrosis is of utmost importance, especially considering the increased incidence of alcoholic and non-alcoholic liver syndromes. In this work, a systematic study is reported to develop a dual function and biocompatible nanoprobe for liver specific diagnostic and therapeutic applications. A polysaccharide polymer, pullulan stabilized iron oxide nanoparticle (P-SPIONs) enabled high liver specificity via asialogycoprotein receptor mediation. Longitudinal and transverse magnetic relaxation rates of 2.15 and 146.91 mM−1 s−1 respectively and a size of 12 nm, confirmed the T2 weighted magnetic resonance imaging (MRI) efficacy of P-SPIONs. A current of 400A on 5 mg/ml of P-SPIONs raised the temperature above 50 °C, to facilitate effective hyperthermia. Finally, a NIR dye conjugation facilitated targeted dual imaging in liver fibrosis models, in vivo, with favourable histopathological results and recommends its use in early stage diagnosis using MRI and optical imaging, and subsequent therapy using hyperthermia.

P-Selectin-Based Dual-Model Nanoprobe Used for the Specific and Rapid Visualization of Early Detection toward Severe Acute Pancreatitis in Vivo.

Identifying severe acute pancreatitis (SAP) as soon as possible is critical for achieving optimal outcomes and saving lives. In this study, a novel P-selectin-targeted, NIR fluorescent dye (Cy 5.5)-labeled dual-modal nanoprobe based on diethylenetriaminepentaacetic chelates (Gd-DTPA-Cy5.5-PsLmAb) was constructed for the bimodal imaging of SAP at the early stage. Gd-DTPA-Cy5.5-PsLmAb was prepared, and its structure was characterized by Fourier transform infrared spectroscopy, UV-vis spectroscopy, and fluorescence spectroscopy, and its stability was evaluated. Biocompatibility was evaluated by the hemolysis and cytotoxicity assays. The enzyme-linked immunosorbent assay was used to detect and evaluate the expression of P-selectin in the peripheral blood of 11 patients with acute pancreatitis (AP) and 5 healthy volunteers. The bimodal imaging ability of Gd-DTPA-Cy5.5-PsLmAb nanoprobes was evaluated via near-infrared fluorescence (NIRF) and magnetic resonance imaging (MRI) in AP animal models in vivo. Gd-DTPA-Cy5.5-PsLmAb showed low toxicity to human embryonic kidney cells (293T cells) and good blood compatibility. The P-selectin levels of humans and rats in the mild acute pancreatitis (MAP)/SAP stage were significantly higher than those in the control group and reached the highest level at the SAP stage. Furthermore, Gd-DTPA-Cy5.5-PsLmAb nanoprobes showed clear NIRF imaging of mouse pancreas at the MAP stage and SAP stage by a fluorescence signal at 6.09 × 108 and 1.95 × 109, respectively. Meanwhile, Gd-DTPA-Cy5.5-PsLmAb nanoprobes also successfully showed the T1-weighted MR signal of rat pancreas at the MAP stage, but Gd-DTPA seldom showed any signal increase at the MAP stage; Gd-DTPA-Cy5.5-PsLmAb and Gd-DTPA could show an increasing MR signal of rat pancreas at the SAP stage. Gd-DTPA-Cy5.5-PsLmAb proved to offer a stronger signal than Gd-DTPA.Our findings indicate that Gd-DTPA-Cy5.5-PsLmAb is an effective and specific MR/NIRF dual nanoprobe for bimodal imaging, providing a promising diagnostic approach for early SAP in clinic.

SPECT/CT imaging of chemotherapy-induced tumor apoptosis using 99mTc-labeled dendrimer-entrapped gold nanoparticles

Non-invasive imaging of apoptosis in tumors induced by chemotherapy is of great value in the evaluation of therapeutic efficiency. In this study, we report the synthesis, characterization, and utilization of radionuclide technetium-99m (99mTc)-labeled dendrimer-entrapped gold nanoparticles (Au DENPs) for targeted SPECT/CT imaging of chemotherapy-induced tumor apoptosis. Generation five poly(amidoamine) (PAMAM) dendrimers (G5.NH2) were sequentially conjugated with 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), polyethylene glycol (PEG) modified duramycin, PEG monomethyl ether, and fluorescein isothiocyanate (FI) to form the multifunctional dendrimers, which were then utilized as templates to entrap gold nanoparticles. Followed by acetylation of the remaining dendrimer surface amines and radiolabeling of 99mTc, the SPECT/CT dual mode nanoprobe of tumor apoptosis was constructed. The developed multifunctional Au DENPs before and after 99mTc radiolabeling were well characterized. The results demonstrate that the multifunctional Au DENPs display favorable colloidal stability under different conditions, own good cytocompatibility in the given concentration range, and can be effectively labeled by 99mTc with high radiochemical stability. Furthermore, the multifunctional nanoprobe enables the targeted SPECT/CT imaging of apoptotic cancer cells in vitro and tumor apoptosis after doxorubicin (DOX) treatment in the established subcutaneous tumor model in vivo. The designed duramycin-functionalized Au DENPs might have the potential to be employed as a nanoplatform for the detection of apoptosis and early tumor response to chemotherapy.

Rhodamine bound maghemite as a long-term dual imaging nanoprobe of adipose tissue-derived mesenchymal stromal cells.

In the last few years, magnetically labeled cells have been intensively explored, and non-invasive cell tracking and magnetic manipulation methods have been tested in preclinical studies focused on cell transplantation. For clinical applications, it is desirable to know the intracellular pathway of nanoparticles, which can predict their biocompatibility with cells and the long-term imaging properties of labeled cells. Here, we quantified labeling efficiency, localization, and fluorescence properties of Rhodamine derivatized superparamagnetic maghemite nanoparticles (SAMN-R) in mesenchymal stromal cells (MSC). We investigated the stability of SAMN-R in the intracellular space during a long culture (20days). Analyses were based on advanced confocal microscopy accompanied by atomic absorption spectroscopy (AAS) and magnetic resonance imaging. SAMN-R displayed excellent cellular uptake (24h of labeling), and no toxicity of SAMN-R labeling was found. 83% of SAMN-R nanoparticles were localized in lysosomes, only 4.8% were found in mitochondria, and no particles were localized in the nucleus. On the basis of the MSC fluorescence measurement every 6days, we also quantified the continual decrease of SAMN-R fluorescence in the average single MSC during 18days. An additional set of analyses showed that the intracellular SAMN-R signal decrease was minimally caused by fluorophore degradation or nanoparticles extraction from the cells, main reason is a cell division. The fluorescence of SAMN-R nanoparticles within the cells was detectable minimally for 20days. These observations indicate that SAMN-R nanoparticles have a potential for application in transplantation medicine.

Folic acid-conjugated GdPO4:Tb3+@SiO2 Nanoprobe for folate receptor-targeted optical and magnetic resonance bi-modal imaging

Both fluorescent and magnetic nanoprobes have great potential applications for diagnostics and therapy. In the present work, a folic acid-conjugated and silica-modified GdPO4:Tb3+ (GdPO4:Tb3+@SiO2-FA) dual nanoprobe was strategically designed and synthesized for the targeted dual-modality optical and magnetic resonance (MR) imaging via a facile aqueous method. Their structural, optical, and magnetic properties were determined using transmission electron microscopy (TEM), X-ray diffraction (XRD), Fourier transform infrared (FTIR), ultraviolet-visible spectra (UV-Vis), photoluminescence (PL), and superconducting quantum interference device (SQUID). These results indicated that GdPO4:Tb3+@SiO2-FA were uniform monodisperse core-shell structured nanorods (NRs) with an average length of ~200 nm and an average width of ~25 nm. The paramagnetic property of the synthesized GdPO4:Tb3+@SiO2-FA NRs was confirmed with its linear hysteresis plot (M-H). In addition, the NRs displayed an obvious T1-weighted effect and thus it could potentially serve as a T1-positive contrast agent. The NRs emitted green lights due to the 5D4 → 7F5 transition of the Tb3+. The in vitro assays with NCI-H460 lung cancer cells and human embryonic kidney cell line 293T cells indicated that the GdPO4:Tb3+@SiO2-FA nanoprobe could specifically bind the cells bearing folate receptors (FR). The MTT assay of the NRs revealed that its cytotoxicity was very low. Further in vivo MRI experiments distinctively depict enhanced anatomical features in a xenograft tumor. These results suggest that the GdPO4:Tb3+@SiO2-FA NPs have excellent imaging and cell-targeting abilities for the folate receptor-targeted dual-modality optical and MR imaging and can be potentially used as the nanoprobe for bioimaging.

Dual Nanoprobe for Single Cell Viability Detection: Method Characterization

This paper presents characterization results of the dual nanoprobe technique for single cell viability detection. Characterization is one of the steps in improving single cell viability detection technique in term of dual nanoprobe sensitivity, design and measurement configuration. The characterizations were focused on improving dual nanoprobe sensitivity and design by studying the effect of different material types, cross sections and measurement configuration, i.e. penetration depth and the gap of the dual nanoprobe on the measurement result. From the findings, the most preferred material is Tungsten and different cross section shapes do not give significant differences in dual nanoprobe sensitivity. It was also found that the current flow increases significantly with deeper penetration depth and narrower probes gap. Therefore, penetration depth and gap need to be constant during measurement in order to get reliable single cell viability detection result. The dual nanoprobe also has the potential to be used in single cell surgery, single cell thermal measurement, single cell drug delivery, and early disease detection applications.

Rigid and Conductive Dual Nanoprobe for Single Cell Analysis

Electrical property characterization of a single cell can be used to infer about its physiological condition, e.g. cell viability. Due to that, a dual nanoprobe-microfluidic system for electrical properties measurement of single cells has been proposed. This paper is concerned about the mechanical and electrical characterizations of the dual nanoprobe. Electrical and mechanical characterizations were conducted to measure the resistance and the strength of the dual nanoprobe for five different metals i.e. Aluminium, Copper, Silver, Tungsten, and Zinc using finite element approach. From the findings, Tungsten’s nanoprobe has the highest strength while the resistance values for the five materials are not significantly different. Therefore, Tungsten is selected as the most recommended metal for the dual nanoprobe. We also performed single cell electrical measurement to test the functionality of the sensor. This work provides general information of the nanoprobe which can be used as a framework in other applications involving Nano devices i.e. cell surgery and drug delivery.

Analysis of Normal and Infected Bio-Cell Using Dual Nanoprobe

The objective of this paper is to analyze the yeast cell, liver cell and blood cell under both infected and healthy conditions by applying electrical signal through dual nanoprobe from source to cell. Knowledge of nanoprobe based bio-cell analysis can be used to differentiate infected cells from healthy ones, since their electrical behaviour is different. The voltage can be applied either inside the cytoplasm penetrating the cell wall, or at the outer surface of cell membrane. In this paper, the simulation has been carried out by applying voltage inside the cytoplasm using ABAQUS 6.10 CAE, powerful finite element software and the results obtained from simulation shows current flow healthy yeast and dead yeast cells are 1·9nA and 34pA respectively whereas the value of the current measured for the leukaemia affected blood cell is 21·2nA, being 2% less than the White Blood Cell (WBC). Since the conductivity of the cytoplasm of the healthy cell is theoretically higher than that of dead or infected cell, which is verified by simulation. The results from simulation show that the measured cell current from liver tumour cell is 2-7 times larger than healthy liver cell including membrane as it is supposed to be since the conductivity of cell including cell membrane is theoretically greater for dead or infected cell than healthy cell.

Instantaneous and Quantitative Single Cells Viability Determination Using Dual Nanoprobe Inside ESEM

We performed single pulses current measurement on single cells using dual nanoprobe through environmental scanning electron microscope nanomanipulator system. The ability to characterize the electrical property of single cells can be used as a novel method for cell viability detection in quantitative and instantaneous manners. The nanoprobe was successfully fabricated using focused ion beam tungsten deposition and etching processes. The characteristics of the nanoprobe were examined from the energy dispersion spectrometry and noise analyses. In this paper, for the first time, the electrical property of single cells under their native condition was presented. In order to apply this method for cell viability detection, two types of cells were used, i.e., dead cells and live cells. The results showed that there is a significant difference on the electrical measurement data between dead and live cells.

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