Jesse Trekker

Temperature Determination of Resonantly Excited Plasmonic Branched Gold Nanoparticles by X‐ray Absorption Spectroscopy

The fields of bioscience and nanomedicine demand precise thermometry for nanoparticle heat characterization down to the nanoscale regime. Since current methods often use indirect and less accurate techniques to determine the nanoparticle temperature, there is a pressing need for a direct and reliable element-specific method. In-situ extended X-ray absorption fine structure (EXAFS) spectroscopy is used to determine the thermo-optical properties of plasmonic branched gold nanoparticles upon resonant laser illumination. With EXAFS, the direct determination of the nanoparticle temperature increase upon laser illumination is possible via the thermal influence on the gold lattice parameters. More specifically, using the change of the Debye-Waller term representing the lattice disorder, the temperature increase is selectively measured within the plasmonic branched nanoparticles upon resonant laser illumination. In addition, the signal intensity shows that the nanoparticle concentration in the beam more than doubles during laser illumination, thereby demonstrating that photothermal heating is a dynamic process. A comparable temperature increase is measured in the nanoparticle suspension using a thermocouple. This good correspondence between the temperature at the level of the nanoparticle and at the level of the suspension points to an efficient heat transfer between the nanoparticle and the surrounding medium, thus confirming the potential of branched gold nanoparticles for hyperthermia applications. This work demonstrates that X-ray absorption spectroscopy-based nanothermometry could be a valuable tool in the fast-growing number of applications of plasmonic nanoparticles, particularly in life sciences and medicine.

Intraarterial route increases the risk of cerebral lesions after mesenchymal cell administration in animal model of ischemia

Mesenchymal stem cells (MSCs) are a promising clinical therapy for ischemic stroke. However, critical parameters, such as the most effective administration route, remain unclear. Intravenous (i.v.) and intraarterial (i.a.) delivery routes have yielded varied outcomes across studies, potentially due to the unknown MSCs distribution. We investigated whether MSCs reached the brain following i.a. or i.v. administration after transient cerebral ischemia in rats, and evaluated the therapeutic effects of both routes. MSCs were labeled with dextran-coated superparamagnetic nanoparticles for magnetic resonance imaging (MRI) cell tracking, transmission electron microscopy and immunohistological analysis. MSCs were found in the brain following i.a. but not i.v. administration. However, the i.a. route increased the risk of cerebral lesions and did not improve functional recovery. The i.v. delivery is safe but MCS do not reach the brain tissue, implying that treatment benefits observed for this route are not attributable to brain MCS engrafting after stroke.

Synthetic Antiferromagnetic Nanoparticles as Potential Contrast Agents in MRI

We present the top-down synthesis of a novel type of MRI T2 contrast agent with great control over size and shape using a colloidal lithography technique. The resulting synthetic antiferromagnetic nanoparticles (SAF-NPs) yield improved relaxivities compared to superparamagnetic iron oxide alternatives (SPIONs). For T2 weighted imaging, the outer sphere relaxation theory has shown that the sensitivity of a T2 contrast agent is dependent on the particle size with an optimal size that exceeds the superparamagnetic limit of SPIONs. With the use of the interlayer exchange coupling effect, the SAF-NPs presented here do not suffer from this limit. Adjusting the outer sphere relaxation theory for spherical particles to SAF-NPs, we show both theoretically and experimentally that the SAF-NP size can be optimized to reach the r2 maximum. With measured r2 values up to 355 s(-1) mM(-1), our SAF-NPs show better performance than commercial alternatives and are competitive with the state-of-the-art. This performance is confirmed in an in vitro MRI study on SKOV3 cells.

PEGylated lipids impede the lateral diffusion of adsorbed proteins at the surface of (magneto)liposomes

Protein binding to nanoparticles is a crucial issue in biomedicine, as it triggers their clearance from the bloodstream after intravenous injection. Many techniques are available for measuring strong protein binding interactions, but weak dynamic interactions are more difficult to assess. To tackle the latter problem, in the present work, cytochrome c was chosen as a representative model of a water-soluble protein and the adsorbing particulates were either small unilamellar phospholipid vesicles or 14 nm diameter solid superparamagnetic iron oxide cores onto which a phospholipid bilayer was strongly chemisorbed (so-called magnetoliposomes). Incorporation of cytochrome c oxidase into the phospholipid bilayer allowed the association of cytochrome c with the surface of the particles to be measured with high sensitivity by VIS-spectrophotometry. The impact of enzyme density as well as some of the physical features of the PEG corona (degree of PEGylation and PEG chain length) adjacent to the surface of the lipid structures on the overall kinetics was also investigated.

Synthesis of PEGylated Magnetic Nanoparticles With Different Core Sizes

Tailoring the properties of superparamagnetic nanoparticles (MNPs) is essential for various nano-based biological applications. Having control over the properties of the MNPs permits a maximum flexibility. Starting from monodisperse iron oxide MNPs produced by thermal decomposition, we report on the optimization and characterization of a first and second seed mediated growth step by varying the surfactant amount and by optimizing the heating steps. We demonstrate the ability to gradually increase the size of crystalline MNPs from 6 over 9 to 12 nm with an improving monodispersity as demonstrated by Transmission Electron Microscopy, Dynamic Light Scattering and X-ray diffraction. The magnetic properties of the MNPs, studied by Vibrating Sample Magnetometry, were in concert with their size increase. We also show the functionalization of these particles with polyethylene glycolated silanes, to render the MNPs stable in water. Different characterization techniques, namely Transmission Electron Microscopy, Dynamic Light Scattering, Fourier-transform InfraRed, Thermo gravimetric analysis and X-ray Photoelectron Spectroscopy, confirmed the successful engraftment of the silanes on the MNPs surface. In conclusion, the proposed route of step-wise synthesis in combination with silane functionalization allows fine tuning the physical properties of iron oxide MNPs for applications in an aqueous environment.

Monitoring the Bystander Killing Effect of Human Multipotent Stem Cells for Treatment of Malignant Brain Tumors

Tumor infiltrating stem cells have been suggested as a vehicle for the delivery of a suicide gene towards otherwise difficult to treat tumors like glioma. We have used herpes simplex virus thymidine kinase expressing human multipotent adult progenitor cells in two brain tumor models (hU87 and Hs683) in immune-compromised mice. In order to determine the best time point for the administration of the codrug ganciclovir, the stem cell distribution and viability were monitored in vivo using bioluminescence (BLI) and magnetic resonance imaging (MRI). Treatment was assessed by in vivo BLI and MRI of the tumors. We were able to show that suicide gene therapy using HSV-tk expressing stem cells can be followed in vivo by MRI and BLI. This has the advantage that (1) outliers can be detected earlier, (2) GCV treatment can be initiated based on stem cell distribution rather than on empirical time points, and (3) a more thorough follow-up can be provided prior to and after treatment of these animals. In contrast to rodent stem cell and tumor models, treatment success was limited in our model using human cell lines. This was most likely due to the lack of immune components in the immune-compromised rodents.

Easy and efficient cell tagging with block copolymers based contrast agents for sensitive MRI detection in vivo.

Superparamagnetic iron oxide nanoparticles (MNPs) together with magnetic resonance imaging (MRI) are the preferred tools for monitoring the fate and biodistribution of administered cells in stem cell therapy studies. Commercial MNPs need transfection agents and long incubation times for sufficient cell labeling and further in vivo cell detection. In this work, we have synthesized MNPs coated with pluronic F127 and tetronic 908, and validated their applicability as contrast agents for MRI cell detection on two different cell types: rat mesenchymal stem cells (MSCs) and multipotent neural progenitor cell line from mice (C17.2). No transfection agent was needed for a complete MNP internalization, and the uptake was only dependent on MNP concentration in medium and limited on the incubation time. By combining in vivo MRI and ex vivo histology microscopy, we have demonstrated the MRI signal detected corresponded exclusively to labeled cells and not to free particles. Pluronic F127- and tetronic 908-coated MNPs represent promising contrast agents for stem cell tracking due to their ease of use in preparation, their efficiency for cell labeling, and their high sensitivity for in vivo cell detection.

Enzyme conjugation and biosensing with quantum dots: A photoluminescence study

In this paper we investigate the enzyme conjugation of CdSe/ZnS core/shell quantum dots (QDs) by means of their photoluminescent properties, as well as the influence of the enzymatic reaction on these properties. The QDs were first transferred from toluene to water by encapsulation in poly(ethylene glycol)-phospholipid micelles. The water solubilized QDs were bioconjugated with the enzymes glucose oxidase (GOX) and horseradish peroxidase (HRP). For covalent coupling the crosslinkers 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and hydroxysulfosuccinimide (sulfo-NHS) were used. Photoluminescence (PL) measurements showed that EDC had a large effect on the PL intensity. Also, the PL intensity increased in PBS buffer as compared to water. The enzyme conjugation caused a noticeable shift in the PL peak wavelength, but had a minor effect on the PL intensity. Sensing experiments confirmed the well-known quenching of PL by H2O2. Similarly, the PL intensity was sensitive to glucose in the mM range in the presence of GOX. This can be understood since the conversion of glucose by GOX yields H2O2 as a by-product. Measurements of the quantum yield (QY) showed increased values for QD solutions treated with EDC. The QY decreased with increasing glucose concentration in the presence of GOX. The change in QY was accompanied by a change in pH, suggesting a correlation between pH and QY.

Assessment of bystander killing-mediated therapy of malignant brain tumors using a multimodal imaging approach

IntroductionIn this study, we planned to assess if adult stem cell-based suicide gene therapy can efficiently eliminate glioblastoma cells in vivo. We investigated the therapeutic potential of mouse Oct4− bone marrow multipotent adult progenitor cells (mOct4− BM-MAPCs) in a mouse glioblastoma model, guided by multimodal in vivo imaging methods to identify therapeutic windows.MethodsMagnetic resonance imaging (MRI) of animals, wherein 5 × 105 syngeneic enhanced green fluorescent protein-firefly luciferase-herpes simplex virus thymidine kinase (eGFP-fLuc-HSV-TK) expressing and superparamagnetic iron oxide nanoparticle labeled (1 % or 10 %) mOct4− BM-MAPCs were grafted in glioblastoma (GL261)-bearing animals, showed that labeled mOct4− BM-MAPCs were located in and in close proximity to the tumor. Subsequently, ganciclovir (GCV) treatment was commenced and the fate of both the MAPCs and the tumor were followed by multimodal imaging (MRI and bioluminescence imaging).ResultsIn the majority of GCV-treated, but not phosphate-buffered saline-treated animals, a significant difference was found in mOct4− BM-MAPC viability and tumor size at the end of treatment. Noteworthy, in some phosphate-buffered saline-treated animals (33 %), a significant decrease in tumor size was seen compared to sham-operated animals, which could potentially also be caused by a synergistic effect of the immune-modulatory stem cells.ConclusionsSuicide gene therapy using mOct4− BM-MAPCs as cellular carriers was effective in reducing the tumor size in the majority of the GCV-treated animals leading to a longer progression-free survival compared to sham-operated animals. This treatment could be followed and guided noninvasively in vivo by MRI and bioluminescence imaging. Noninvasive imaging is of particular interest for a rapid and efficient validation of stem cell-based therapeutic approaches for glioblastoma and hereby contributes to a better understanding and optimization of a promising therapeutic approach for glioblastoma patients.