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In C and D. Sagittal MR images taken at different depths of the same knee at different time points. Alginate constructs are crosslinked with 2mM gadolinium In C the rat knee right after injection of MSC-alginate beads, the beads, visible as white dots, are clustered mostly at the suprapatellar joint space. We can see that the injected beads have moved and can be found throughout the joint. The microbeads were labeled with Gadolinium, this way we could follow them on MRI.

Previously the group - including dr. Haeck and dr. Nieuwstadt had developed a robust quantitative MR imaging tools for evaluation construct integrity. And as seen in the images we could still detect the beads after weeks of in vivo. It causes pain and impairs movement, thereby reducing quality of life. To date there is no treatment to stop or cure the disease. MSCs are very promising as cell therapy, because they have immunomodulatory and trophic capacities that can influence patho physiological processes and tissue repair.

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In this study we intend to develop alginate microcapsules with MSCs for intra-articular injection as a durable treatment for osteoarthritis. We will optimize the composition of the microcapsules to achieve prolonged survival of the cells and therapeutic effect by the therapeutic properties of the MSCs.

The aim of the ongoing studies is to evaluate the effect of the MSC microcapsules intra-articular considering their long-term integrity and their effect on the environment. We developed robust quantitative MR imaging tools for evaluation of cell survival and construct integrity. Encapsulated MSCs in microcapsules retained their therapeutic properties, indicating that we can prolong survival and function of MSCs.

In vivo characterization of alginate constructs by MRI. A In vivo quantitative MRI data of subcutaneous implanted alginate constructs in a rat. Alginate constructs are crosslinked with 20mM gadolinium resulting in low T1 times. B Identification of presence and localization of alginate constructs after intraarticular injection.

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Upon rupture of plaques and subsequent thrombus formation, a stroke or myocardial infarct can follow. Non-invasive visualization of atherosclerotic plaques remains a challenge, especially in coronary arteries. Moreover, the ability to differentiate between phenotypically rupture prone and more stable plaques is a major objective in current research. As inflammation is crucial in the onset-and progression of atherosclerosis, we aim to visualize inflammation by targeting macrophages, the most common inflammatory cell in atherosclerotic plaques. Leukocyte Function Associated antigen 1 LFA1 is expressed on macrophages and is involved in macrophage recruitment to sites of infection and inflammation.

Further ex vivo studies confirmed these findings, indicating that the in vivo signal indeed stems from plaque and not surrounding tissue. Future studies will elucidate which stages of disease can be discriminated using Danbirt, and if plaques can be differentiated according to plaque phenotype. In complementary studies we are using different tracers to explore the possibility to visualize different macrophage subtypes. Information on the pro-or anti-inflammatory phenotype of macrophages might yield more information on plaque status.

Our current and expected future results will aid clinicians in the evaluation of disease presence, stage, plaque status, and yield an indication of treatment effect. Signal is visible from the aortic arch. Right image: ex vivo autoradiograph of excised arteries. Signal originates from plaques located in the aortic arch and carotid bifurcations. However, this therapy is primarily focused on external radiation of local tumors and not effective when used in metastasizing cases.

To treat metastatic sites more effectively, systemic radionuclide therapy was developed. Relative recent findings showed that radionuclides can be specifically targeted to tumors. A therapy already used in the clinic exploits the somatostatin receptors, over-expressed in neuro-endocrine. This therapy shows great potential, considering a customized setup where the efficiency is enhanced, targeting is optimized and generating a system to change the target or radionuclide relatively easy opens up a whole new field of investigation.

Nano-carriers have been proven to be viable candidates for customizable systems, especially polymeric vesicles polymersomes. Alpha-irradiation setup. Coverslips are put on mylar to cover the cells. Using an old microscope the cells are irradiated by the Am source by placing the source very close to the cells.

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After 3 minutes of irradiation the cells are put back in to culture medium and are fixated after hours for immunohistochemical staining. Anti-tumor therapy can be maximized when the genetic background of certain tumors are taken in to account. Ionizing radiation can cause all kinds of DNA damage, including double stranded breaks DSBs , which are the most harmful. Alpha irradiation has proven to be of exceptional use to study DSBs induced by ionizing irradiation. By introducing straight paths of DSBs in the nucleus of the cell endogenous induced breaks are discriminated from induced breaks by ionizing irradiation.

Using this technique we can investigate the DDR in great detail using immunofluorescence. However, this technique requires non-typical culture conditions and was only possible for fixed time points. This project improved the technique through optimizing the irradiation of cells from another perspective, using mylar as a protective barrier which alpha-particles can pass through picture.

The improved method of irradiation allows normal culture condition and in addition Super Resolution imaging of alpha particle induced damage which can greatly enhance the knowledge of DSBs and the DDR. The main intent of the whole consortium is the development of new theranostic agents that can play a role in the fight against cancer. The minimal involvement of off target organs and the high retention in the tumor suggests that P94 has the potential to be used as a carrier for therapeutic radionuclides.

Pluronics are amphipathic tri-block copolymers, commonly used in industrial application, cosmetics and pharmaceutical preparations. Modified and radiolabeled Pluronics can be used as theranostic carriers to deliver drugs and imaging agents at the tumor site. We compared the biodistribution of P94 following two different injection routes: intratumoral IT and intravenous IV.

Results panel A showed that PIn-DTPA had little accumulation in the tumor and elevated accumulation in the liver but also little accumulation in off target tissue after IV injection. Furthermore the improved retention of radioactivity seemed to be specifically mediated via the P94 copolymer panel B as IT injection of In-DTPA resulted in fast excretion of radioactivity. The superior soft tissue contrast of the MR image aids in the delineation of the organs, whereas the SPECT signal allows for accurate quantification of the administered radiopharmaceutical.

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These two properties combined offer great potential for research and development of theranostic compounds. Accurate quantification, relying on adequate attenuation correction, is of key importance to be a viable alternative for ex vivo biodistribution studies. A CT scan provides accurate means for this correction, for MR this correction is more difficult and involves segmentation of the MR image in different tissue classes.

Dosimetry studies are performed to assess the radiation dose and expediency of a theranostic compound. To determine the exact amount of radioactivity in each organ, necessary to calculate the radiation doses, accurate. A second part of this project will focus on performance characterization of the SPECT subsystem in terms of resolution, image contrast, sensitivity and partial volume effects.

PC3 tumor bearing mouse imaged 24h after injection with In-Sarabesin3. The uptake of In-Sarabesin3 is well confined within the tumor left. Additionally some activity in the kidney cortex remains. There is an enormous interest in the role of macrophages polarization in tissue maintenance, repair and breakdown. Although in vivo imaging of macrophages can potentially elucidate how chronic diseases initiate and evolve, current work also clearly shows a weakness, namely lack of knowledge on macrophage polarization.

Macrophages are present in virtually all tissues of the body. There are tissue-specific macrophages of bone, central nervous system, connective tissue and many more. When macrophages become activated, they can: 1 act as immune effector cells, 2 assist in tissue development, and 3 participate in tissue remodeling. Depending on their functional state, macrophages can facilitate or prevent disease. The amount of RA measured fluctuates in time Fig. In order to assess the whether the PET signal is a measure for the level of macrophage-specific activation in the osteoarthritic knee, knees have been resected at different time points too and processed for histological analysis.

Currently, historical analysis of the knees is being performed and related to the imaging results obtained. We are also evaluating novel tracers for their use as specific markers for activated macrophages. The first tracer tested was the in-house available InOctreoscan. This tracer binds to several subtypes of the somatostatin receptor SSTR as reported before.

We subsequently confirmed that the SSTR 2 , is expressed on pro-inflammatory macrophages, through rt-pcr analysis and tracer uptake experiments In vivo studies with In-Octreoscan, unfortunately revealed that the sensitivity of this SPECT tracer is not high enough.

Mice injected with collagenase in the intra-articular space were imaged at different time points days after induction of OA. PET scans show radioactivity RA present in. Kristell Chatalic obtained her PhD in on a topic at the interface of radiochemistry, molecular imaging and radionuclide therapy under the supervision of Prof. Otto C. Boerman Radboud University Medical Center. The third section focuses on PSMA targeting.

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This Nanobody was engineered with a C-terminal cysteine or site-specific coupling of a chelator. In Chapter 3. In this chapter we investigated a strategy to decrease renal tracer uptake.

The first section of this thesis presents the theranostic concept in nuclear medicine. Chapter 1. Strategies to improve applications of radiolabeled peptides for imaging and therapy are discussed in Chapter 1.